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  • Immunohistochemistry  (72)
  • Saccharomyces cerevisiae  (60)
  • Springer  (132)
  • American Chemical Society (ACS)
  • American Institute of Physics (AIP)
  • 2015-2019
  • 1995-1999  (132)
  • 1935-1939
  • 1925-1929
  • 2015
  • 1995  (132)
Collection
Keywords
Publisher
Years
  • 2015-2019
  • 1995-1999  (132)
  • 1935-1939
  • 1925-1929
Year
  • 1
    Electronic Resource
    Electronic Resource
    Peptidergic innervation of leg muscles of the cockroach, Periplaneta americana (L.), and a possible role in modulation of muscle contraction (1995)
    Springer
    Journal of comparative physiology 176 (1995), S. 425-435 
    Details
    ISSN: 1432-1351
    Keywords: FaRPs ; FMRFamide Nervous system Skeletal muscle ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract FMRFamide-related peptides of insects are particularly important because of their possible function as neurohormones and neuromodulators on a wide variety of tissues. Part of this study was an investigation of the immunofluorescent staining of motor nerves which arise in the metathoracic ganglion, examined in wholemount using an antiserum that recognizes extended -RFamide peptides (generally recognized to be of the FMRFamide family). This antiserum revealed immunochemical staining of numerous cell bodies in the metathoracic ganglion and of axons in peripheral nerve 5, a large nerve which contains both motor and sensory fibres. Axons staining positive for FMRFamide-related peptides were traced in nerve 5 as far as the femur-tibia joint, and into the first (sensory-motor) and third (motor only) ramus of nerve 5. Reverse-phase HPLC with radioimmunoassay revealed a peak of FMRFamide-related peptide activity in nerve 5 that was coincident with a peak found when thoracic ganglia were processed in the same fashion. A physiological assay was devised to test the ability of various non-native peptides to alter the characteristics of contraction of skeletal muscles of the legs. Using neurally evoked contractions of coxal depressor muscles of the metathoracic leg it was determined that several non-native peptides could potentiate muscle contractions. The results of this study suggest that muscles of the legs receive innervation by identifiable, FMRFamide-related peptide-containing neurons and that the release of peptide(s) at the muscle may be yet another method of modulating the mechanics of muscle contraction.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00219067
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  • 2
    Electronic Resource
    Electronic Resource
    Octopaminergic neurons in the locust brain: morphological, biochemical and electrophysiological characterisation of potential modulators of the visual system (1995)
    Springer
    Journal of comparative physiology 177 (1995), S. 611-625 
    Details
    ISSN: 1432-1351
    Keywords: Cobalt staining ; Gas chromatography-mass spectrometry ; Immunohistochemistry ; Insect ; Neuromodulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The two Protocerebral-Medulla 4 neurons (PM4a and b) in the locust brain have adjacent cell bodies in the medial deutocerebrum. They project through the posterior protocerebrum, forming limited arborisations en route, and enter the lobula and medulla of the ipsilateral optic lobe, where they form extensive, overlapping arborisations. The PM4a and b neurons are octopamine immunoreactive. Their octopamine content (approximately 25 pg per cell) is confirmed by gas chromatography-mass spectrometry; each cell contains approximately 25 pg p-octopamine. Simultaneous intracellular recording from exposed PM4a and b cell bodies reveals that the two cells are physiologically indistinguishable. They receive multimodal sensory inputs. Tactile/mechanosensory stimuli to much of the animal's body and head, acoustic stimuli, and simple visual stimuli all give rise to e.p.s.p.s and action potentials in the PM4 cell body. Simultaneous recording from the cell body in the deutocerebrum and the axon in the lobula demonstrates that action potentials are predominantly initiated in the deutocerebrum and propagate centrifugally, towards the optic lobe. Occasionally, bright light flashes will initiate an action potential in the axon in the optic stalk, which probably propagates bidirectionally: centripetally to the cell body, and centrifugally into the optic lobe. The extensive arborisations in the lobula and medulla are therefore likely to be sites of octopamine release. Because PM4 neurons are octopaminergic, project to the optic lobe, and receive modalities of sensory input known to dishabituate the Descending Contralateral Movement Detector (DCMD) visual interneuron, it is proposed that PM4 neurons are neuromodulatory — mediating dishabituation or arousal of the visual system.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00207190
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  • 3
    Electronic Resource
    Electronic Resource
    Expression of a yeast gene can be blocked by insertion of short yeast DNA fragments between a UAS and the TATA box (1995)
    Springer
    Current genetics 27 (1995), S. 387-389 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; URS ; FBP1 Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have constructed a plasmid, pOV10, which facilitates the introduction of putative upstream activating sequences (UAS) or upstream repressing sequences (URS) from yeast genes into plasmids containing CYC1-lacZ fusions. We have observed that the insertion of yeast sequences from 155 to 195 bp between the UAS and the TATA box of a CYC1-lacZ fusion gene can block β-galactosidase expression. It is suggested that this block is related to the formation of nucleosomes on the DNA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00352109
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  • 4
    Electronic Resource
    Electronic Resource
    NCA3, a nuclear gene involved in the mitochondrial expression of subunits 6 and 8 of the Fo-F1 ATP synthase of S. cerevisiae (1995)
    Springer
    Current genetics 27 (1995), S. 409-416 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mitochondrial synthesis ; Nuclear control ; F1Fo-ATPase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Respiratory-competent nuclear mutants have been isolated which presented a cryosensitive phenotype on a non-fermentative carbon source, due to a dysfunctioning of the mitochondrial F1-Fo ATP synthase which results from a relative defect in subunits 6 and 8 of the Fo sector. Both proteins are mtDNA-encoded, but the defect is due to the simultaneous presence of a mutation in two unlinked nuclear genes (NCA2 and NCA3, for Nuclear Control of ATPase) promoting a modification of the expression of the ATP8-ATP6 co-transcript (formerly denoted AAP1-OLI2). This co-transcript matures at a unique site to give two co-transcripts of 5.2 and 4.6 kb in length: in the mutant, the 5.2-kb co-transcript was greatly lowered. NCA3 was isolated from a wild-type yeast genomic library by genetic complementation. The level of the 5.2-kb transcript, like the synthesis of subunits 6 and 8, was partly restored in the transformed strain. A 1011-nucleotide ORF was identified that encodes an hydrophilic protein of 35417 Da. Disruption of chromosomal DNA within the reading frame promoted a dramatic decrease of the 5.2-kb mRNA but did not abolish the respiratory competence of a wild-type strain. NCA3 is located on chromosome IV and produces a single 1780-b transcript.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00311209
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  • 5
    Electronic Resource
    Electronic Resource
    Failure to detect an antimutator phenotype following disruption of the Saccharomyces cerevisiae DDR48 gene (1995)
    Springer
    Current genetics 27 (1995), S. 496-500 
    Details
    ISSN: 1432-0983
    Keywords: Antimutator ; DDR48 ; Saccharomyces cerevisiae ; Spontaneous mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The antimutator phenotype, reportedly conferred by disruption of the Saccharomyces cerevisiae DDR48 gene, was suggested to affect only a specific spontaneous mutational pathway. We attempted to identify the types of mutation that are DDR48-dependent by determining the specificity of the ddr48 antimutator. However, disruption of DDR48 did not decrease the rates of spontaneous forward mutation in a plasmid-borne copy of the yeast SUP4-o gene, the reversion or suppression of the lys2–1 allele, or forward mutation at the CAN1 locus. Interestingly, the latter gene had been reported previously to be subject to the antimutator effect. DNA sequence analysis of spontaneous SUP4-o mutations arising in DDR48 and ddr48 backgrounds provided no evidence for a reduction in the rates of individual mutational classes. Thus, we were unable to verify that disruption of DDR48 causes an antimutator phenotype.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00314438
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  • 6
    Electronic Resource
    Electronic Resource
    Purification and binding properties of the Mal63p activator of Saccharomyces cerevisiae (1995)
    Springer
    Current genetics 27 (1995), S. 509-516 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Maltose fermentation ; MAL63 ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mal63p is a transcriptional activator for maltose fermentation in Saccharomyces cerevisiae. We have purified it to homogeneity from a yeast strain in which the MAL63 gene is under the control of the GAL1–GAL10 promoter. Purification included fractionation of a whole-cell extract by ion-exchange chromatography, chromatography using both non-specific DNA-affinity (calf thymus), and sequence-specific DNA-affinity chromatography. Mal63p activity was assayed by its binding to a fragment of the MAL61–MAL62 promoter, using both filter-binding and electrophoretic-mobility shift assays. DNase-I footprinting identified a new binding site (site 3) between the two previously known sites (sites 1 and 2). Mal63p is a dimer, and methylation-protection experiments identify the recognition motif as: c/a GC N9 c/a GC/g.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00314440
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  • 7
    Electronic Resource
    Electronic Resource
    The PSO4 gene of S. cerevisiae is important for sporulation and the meiotic DNA repair of photoactivated psoralen lesions (1995)
    Springer
    Current genetics 27 (1995), S. 207-212 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; pso4-1 mutant Sporulation ; DNA repair ; Meiotic recombination Induced mutagenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have evaluated the effect of the Saccharomyces cerevisiae pso4-1 mutation in sporulation and DNA repair during meiosis. We have found that pso4-1 cells were arrested in an early step of meiosis, before premeiotic DNA synthesis, and hence did not produce spores. These results suggest that the PSO4 gene may act at the start point of the cell cycle, as do some SPO and CDC genes. The pso4-1 mutant cells are specifically sensitive to 8-MOP- and 3-CPs-photoinduced lesions, and are found to be severely affected in meiotic recombination as well as impaired in the mutagenic response, as previously described for mitosis. This means that the PSO4 gene is important for the repair 8-MOP-photoinduced lesions, mainly double-strand breaks, and the processing of these lesions into recombinogenic intermediates.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00326150
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  • 8
    Electronic Resource
    Electronic Resource
    Determination of chromosome copy numbers in Saccharomyces cerevisiae strains via integrative probe and blot hybridization techniques (1995)
    Springer
    Current genetics 27 (1995), S. 217-228 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Chromosome copy numbers ; Ploidy probes ; Industrial yeasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Methods have been devised for analyzing chromosome copy numbers in S. cerevisiae strains that may be polyploid or aneuploid, as is apparent in the case of many industrial strains. The initial step involved transformation of a strain with an integrative “ploidy probe” transplacement fragment that enable the copy number of the targeted chromosomal locus to be determined via genomic Southern blotting and quantitative probe hybridization. Dual probe co-hybridization to Southern genomic DNA blots was used to extend such locus copy number determinations to other loci within the same chromosome, thereby screening for internal consistency along the length of the chromosome. This approach was also used to extend the analysis to other chromosomes in the genome. The method was established and verified with euploid series laboratory strains and then used to examine chromosome copy numbers in three industrial strains. One brewing strain apparently contained three copies of the chromosomes tested, whilst another brewing and a baking strain showed evidence of aneuploidy.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00326152
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  • 9
    Electronic Resource
    Electronic Resource
    Cloning and expression of an Aspergillus kawachii endo-1,4-β-xylanase gene in Saccharomyces cerevisiae (1995)
    Springer
    Current genetics 28 (1995), S. 467-473 
    Details
    ISSN: 1432-0983
    Keywords: Aspergillus kawachii ; β-xylanase ; Expression ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract First-strand cDNA was prepared from mRNA isolated from Aspergillus kawachii IFO4308 and the β-xylanase gene (xynC) amplified by using the polymerase chain reaction (PCR) technique. This gene was inserted between the yeast phosphoglycerate kinase (PGK1) gene promoter (PGK1 p) and terminator (PGK1 T) sequences. The PGK1 P-xynC-PGK1 T construct (designated XYN3) was cloned into a multicopy episomal plasmid and the XYN3 gene was expressed in Saccharomyces cerevisiae. Functional β-xylanase (Xyn3) was produced and secreted by the recombinant yeast. Xyn3 was stable between 30 and 50°C, and the optimum temperature and pH were shown to be at 60°C and lower than pH3, respectively. An autoselective fur1::LEU2 XYN3 recombinant strain was developed that allowed β-xylanase production at a level of 300 nkat/ml in a non-selective complex medium.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00310817
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  • 10
    Electronic Resource
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    Strategy for deletion of complete open reading frames in Saccharomyces cerevisiae (1995)
    Springer
    Current genetics 27 (1995), S. 306-308 
    Details
    ISSN: 1432-0983
    Keywords: Gene deletion ; Open reading frame ; Saccharomyces cerevisiae ; Polymerase chain reaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The classical disruption method for yeast genes is by using in vitro deletion of the gene of interest, or of a part of it, with restriction enzymes. We are now routinely using a strategy that takes advantage of polymerase chain reactions (PCRs) which amplify large pieces of DNA. Since this approach results in a complete, precise deletion of the open reading frame, which is replaced by a unique restriction site, the ligated PCR can be used for the insertion of different markers of for two-step gene disruptions without an inserted marker. As we have now used this strategy for the deletion of more than ten genes we have in this report included some hints based on our experience.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00352097
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  • 11
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    Molecular cloning and characterization of a novel gene of Candida albicans, CDR1, conferring multiple resistance to drugs and antifungals (1995)
    Springer
    Current genetics 27 (1995), S. 320-329 
    Details
    ISSN: 1432-0983
    Keywords: Multidrug resistance ; Candida albicans ; Saccharomyces cerevisiae ; ABC transporters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By functional complementation of a PDR5 null mutant of Saccharomyces cervisiae, we have cloned and sequenced the multidrug-resistance gene CDR1 of Candida albicans. Transformation by CDR1 of a PDR5-disrupted host hypersensitive to cycloheximide and chloramphenicol resulted in resistance to cycloheximide, chloramphenicol and other drugs, such as the antifungal miconazole, with collateral hypersensitivity to oligomycin, nystatin and 2,4 dinitrophenol. Our results also demonstrate the presence of several PDR5 complementing genes in C. albicans, displaying multidrug-resistance patterns different from PDR5 and CDR1. The nucleotide sequence of CDR1 revealed that, like PDR5, it encodes a putative membrane pump belonging to the ABC (ATP-binding cassette) superfamily. CDR1 encodes a 1501-residue protein of 169.9 kDa whose predicted structural organization is characterized by two homologous halves, each comprising a hydrophobic region with a set of six transmembrane stretches, preceded by a hydrophilic nucleotide binding fold.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00352101
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  • 12
    Electronic Resource
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    The Saccharomyces cerevisiae HIS3 and LYS2 genes complement the Schizosaccharomyces pombe his5-303 and lys1-131 mutations, respectively: new selectable markers and new multi-purpose multicopy shuttle vectors, pSP3 and pSP4 (1995)
    Springer
    Current genetics 28 (1995), S. 380-383 
    Details
    ISSN: 1432-0983
    Keywords: Autonomously replicating sequence ; Auxotrophy ; Schizosaccharomyces pombe ; Saccharomyces cerevisiae ; Cloning vector ; Selectable marker ; HIS/his ; LYS/lys
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three new S. pombe plasmids are described. Plasmids pSP3 and pSP4 are two Schizosaccharomyces pombe ars1 multicopy vectors with the Saccharomyces cerevisiae HIS3 or LYS2 genes as selectable markers. They complement the S. pombe his5-303 or lys1-131 mutations, respectively. Plasmid pSPars1 is a vector carrying the S. pombe ars1 and a unique NdeI site which allows the introduction of any selectable marker therefore bringing a unified vector backbone for the construction of new S. pombe/S. cerevisiae/E. coli shuttle vectors. These plasmids permit classical molecular genetic techniques to be performed directly.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00326437
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  • 13
    Electronic Resource
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    Calmodulin-dependent protein kinase II and calmodulin are required for induced thermotolerance in Saccharomyces cerevisiae (1995)
    Springer
    Current genetics 27 (1995), S. 190-193 
    Details
    ISSN: 1432-0983
    Keywords: Calmodulin ; Calmodulin-dependent protein kinase II ; Heat shock response ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We show here that yeast mutants lacking calmodulin-dependent protein kinase II fail to fully acquire induced thermotolerance. A similar result was also obtained with mutants depending solely on either the N-terminal half or the C-terminal half of calmodulin. These findings indicate that both calmodulin-dependent protein kinase II and calmodulin are required for induced thermotolerance.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00313434
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  • 14
    Electronic Resource
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    Control of glycolytic gene expression in the budding yeast (Saccharomyces cerevisiae) (1995)
    Springer
    Current genetics 29 (1995), S. 1-9 
    Details
    ISSN: 1432-0983
    Keywords: Glycolysis ; Transcriptional activation ; Saccharomyces cerevisiae ; Chromatin structure ; Glucose induction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00313187
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  • 15
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    A mutant allele of the SUP45 (SAL4) gene of Saccharomyces cerevisiae shows temperature-dependent allosuppressor and omnipotent suppressor phenotypes (1995)
    Springer
    Current genetics 27 (1995), S. 417-426 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Omnipotent suppression ; Nonsense suppression ; SUP45
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using a plasmid-based termination-read-through assay, the sal4-2 conditional-lethal (temperature-sensitive) allele of the SUP45 (SAL4) gene was shown to enhance the efficiency of the weak ochre suppressor tRNA SUQ5 some 10-fold at 30°C. Additionally, this allele increased the suppressor efficiency of SRM2-2, a weak tRNAGln ochre suppressor, indicating that the allosuppressor phenotype is not SUQ5-specific. A sup + sal4-2 strain also showed a temperature-dependent omnipotent suppressor phenotype, enhancing readthrough of all three termination codons. Combining the sal4-2 allele with an efficient tRNA nonsense suppressor (SUP4) increased the temperature-sensitivity of that strain, indicating that enhanced nonsense suppressor levels contribute to the conditional-lethality conferred by the sal4-2 allele. However, UGA suppression levels in a sup + sal4-2 strain following a shift to the non-permissive temperature reached a maximum significantly below that exhibited by a non-temperature sensitive SUP4 suppressor strain. Enhanced nonsense suppression may not therefore be the primary cause of the conditional-lethality of this allele. These data indicate a role for Sup45p in translation termination, and possibly in an additional, as yet unidentified, cellular process.
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    URL: http://dx.doi.org/10.1007/BF00311210
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  • 16
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    Mutants of Saccharomyces cerevisiae sensitive to oxidative and osmotic stress (1995)
    Springer
    Current genetics 27 (1995), S. 427-434 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Oxidative stress ; Osmotic stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Although oxidative stress is involved in many human diseases, little is known of its molecular basis in eukaryotes. In a genetic approach, S. cerevisiae was used to identify elements involved in oxidative stress. By using hydrogen peroxide as an agent for oxidative stress, 34 mutants were identified. All mutants were recessive and fell into 16 complementation groups (pos1 to pos16 for peroxide sensitivity). They corresponded to single mutations as shown by a 2:2 segregation pattern. Enzymes reportedly involved in oxidative stress, such as glucose-6-phosphate dehydrogenase, glutathione reductase, superoxide dismutase, as well as glutathione concentrations, were investigated in wild-type and mutant-cells. One complementation group lacked glucose-6-phosphate dehydrogenase and was shown to be allelic to the glucose-6-phosphate dehydrogenase structural gene ZWF1/MET19. In other mutants all enzymes supposedly involved in oxidative-stress resistance were still present. However, several mutants showed strongly elevated levels of glutathione reductase, gluconate-6-phosphate dehydrogenase and glucose-6-phosphate dehydrogenase. One complementation group, pos9, was highly sensitive to oxidative stress and revealed the same growth phenotype as the previously described yap1/par1 mutant coding for the yeast homologue of mammalian transcriptional activator protein, c-Jun, of the proto-oncogenic AP-1 complex. However, unlike par1 mutants, which showed diminished activities of oxidative-stress enzymes and glutathion level, the pos9 mutants did not reveal any such changes. In contrast to other recombinants between pos mutations and par1, the sensitivity did not further increase in par1 pos9 recombinants, which may indicate that both mutations belong to the same regulating circuit. Interestingly, ten complementation groups were, in parallel, sensitive to osmotic stress, and one mutant allele revealed increased heat sensitivity. Our results indicate that a surprisingly large number of genes seem to be involved in oxidative-stress resistance and a possible overlap exists between osmotic stress and other stress reactions.
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    URL: http://dx.doi.org/10.1007/BF00311211
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  • 17
    Electronic Resource
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    The expression of a specific 2-deoxyglucose-6P phosphatase prevents catabolite repression mediated by 2-deoxyglucose in yeast (1995)
    Springer
    Current genetics 28 (1995), S. 101-107 
    Details
    ISSN: 1432-0983
    Keywords: 2-deoxyglucose ; 2-deoxyglucose-6P phosphatase ; Catabolite repression ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 2-deoxyglucose (2-DOG), a non-metabolize analogue of glucose, is taken up by yeast using the same transporter(s) as glucose and is phosphorylated by hexokinases producing 2-deoxyglucose-6-P. We found that in DOG R yeasts, 2-DOG was not able to trigger glucose repression, even at concentrations of 0.5%. This result suggests that the specific 2-DOG-6P phosphatase, the enzyme responsible for the DOG R phenotype, may be involved in inhibiting the process of catabolite repression mediated by 2-DOG
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    URL: http://dx.doi.org/10.1007/BF00315774
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  • 18
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    Carbon catabolite regulation of transcription of nuclear genes coding for mitochondrial proteins in the yeast Kluyveromyces lactis (1995)
    Springer
    Current genetics 28 (1995), S. 267-273 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Kluyveromyces lactis ; Transcriptional regulation ; Catabolite repression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Promoter regions of the KlQCR7, KlQCR8 and KlCYC1 genes, coding for subunits of the bc 1-complex and cytochrome c respectively, in the shortterm Crabtree-negative yeast Kluyveromyces lactis differ markedly in sequence from their Saccharomyces cerevisiae counterparts. They have, however, conserved very similar configurations of binding-site motifs for various transcription factors known to be involved in global and carbon-source regulation in S. cerevisiae. To investigate the carbon source-dependent expression of these genes in K. lactis, we have carried out medium-shift experiments and determined transcript levels during the shifts. In sharp contrast to the situation in S. cerevisiae, the level of expression in K. lactis is not affected when glucose is added to a non-fermentable carbon-source medium. However, the genes are not constitutively expressed, but become significantly induced when the cells are shifted from glucose to a nonfermentable carbon source. Finally, induction of transcriptional activation does not occur in media containing both glucose and non-femmentable carbon sources.
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    URL: http://dx.doi.org/10.1007/BF00309786
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  • 19
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    Positive and negative elements involved in the differential regulation by heme and oxygen of the HEM13 gene (coproporphyrinogen oxidase) in Saccharomyces cerevisiae (1995)
    Springer
    Current genetics 28 (1995), S. 503-511 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; HEM13 regulation ; Heme and oxygen ; CYP1, ROX1, SSN6, TUP1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Saccharomyces cerevisiae HEM13 gene codes for coproporphyrinogen oxidase (CPO), an oxygen-requiring enzyme catalysing the sixth step of heme biosynthesis. Its transcription is increased 40–50-fold in response to oxygen- or heme-deficiency. We have analyzed CPO activity and HEM13 mRNA levels in a set of isogenic strains carrying single or double deletions of the CYP1 (HAP1), ROX1, SSN6, or TUPI genes. The cells were grown in the presence or absence of oxygen and under heme-deficiency (hem1Δ background). Both Rox1p and Cyp1p partially repressed HEM13 in aerobic heme-sufficient cells, probably in an independent manner. In the absence of heme, Cyp1p activated HEM13 and strongly repressed ROX1, allowing de-repression of HEM13. Cyp1p had no effect on HEM13 expression in anaerobic cells. Deletions of SSN6 or TUP1 dramatically de-repressed HEM13 in aerobic cells. A series of deletions in the HEM13 promoter identified at least four regulatory regions that are required for HEM13 regulation. Two regions, containing motifs similar to the Rox1p consensus sequences, act as repression sites under aerobic growth. The two other sites act as activation sequences required for full induction under oxygen- or heme-deficiency. Taken together, these results suggest that induction of HEM13 occurs in part through relief of repression exerted by Rox1p and Cyp1p, and in part by activation mediated partly by Cyp1p under heme-deficiency and by unknown factors under oxygen-deficiency.
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    URL: http://dx.doi.org/10.1007/BF00518161
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  • 20
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    Characterization of a novel α-amylase from Lipomyces kononenkoae and expression of its gene (LKA1) in Saccharomyces cerevisiae (1995)
    Springer
    Current genetics 28 (1995), S. 526-533 
    Details
    ISSN: 1432-0983
    Keywords: α-Amylase ; Lipomyces kononenkoae ; LKA1 ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A highly active α-amylase (76 250 Da) secreted by the raw starch-degrading yeast Lipomyces kononenkoae strain IGC4052B was purified and characterized. Using high performance liquid chromatography (HPLC), end-product analysis indicated that the L. kononenkoae α-amylase acted by endo-hydrolysis on glucose polymers containing α-1,4 and α-1,6 bonds, producing mainly maltose, maltotriose and maltotetraose. The following NH2-terminal amino acids were determined for the purified enzyme: Asp-Cys-Thr-Thr-Val-Thr-Val-Leu-Ser-Ser-Pro-Glu-Ser-Val-Thr-Gly. The L. kononenkoae α-amylase-encoding gene (LKA1), previously cloned as a cDNA fragment, was expressed in Saccharomyces cerevisiae under the control of the PGK1 promoter. The native signal sequence efficiently directed the secretion of the glycosylated protein in S. cerevisiae. De-glycosylation of the enzyme indicated that post-translational glycosylation is different in S. cerevisiae from that in L. kononenkoae. Zymogram analysis indicated that glycosylation of the protein in S. cerevisiae had a negative effect on enzyme activity. Southern-blot analysis revealed that there is only a single LKA1 gene present in the genome of L. kononenkoae.
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    The upstream region of the isocitrate lyase gene (UPR-ICL) of Candida tropicalis induces gene expression in both Saccharomyces cerevisiae and Escherichia coli by acetate via two distinct promoters (1995)
    Springer
    Archives of microbiology 163 (1995), S. 322-328 
    Details
    ISSN: 1432-072X
    Keywords: Isocitrate lyase ; n-Alkane-utilizable yeast ; Candida tropicalis ; Saccharomyces cerevisiae ; Promoters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The upstream region of the isocitrate lyase gene (UPR-ICL, 1530bp) of an n-alkane-utilizable yeast, Candida tropicalis, induced gene expression in another yeast, Saccharomyces cerevisiae, when the yeasts were grown on acetate. Surprisingly, UPR-ICL displayed the same regulatory function in the bacterium Escherichia coli when grown on acetate. We determined the interesting nucleotide sequence of UPR-ICL. The deletion analysis of UPR-ICL in both cells revealed the presence of two distinct promoters: one was localized at-394 to-379 and regulated gene expression in S. cerevisiae; the other was tocated near the initiation codon and regulated gene expression in E. coli. The two promoter sequences were similar, but not identical to regulatory elements that have been previously reported in S. cerevisiae and E. coli, respectively. Accordingly, the possibility of novel regulatory mechanisms could not be excluded. This is an interesting example of the presence of distinct cis-acting regulatory elements responsible for the induction of gene expression in one gene by acetate in both S. cerevisiae and E. coli. Preservation of such promoters through evolution is also discussed.
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    The upstream region of the isocitrate lyase gene (UPR-ICL) of Candida tropicalis induces gene expression in both Saccharomyces cerevisiae and Escherichia coli by acetate via two distinct promoters (1995)
    Springer
    Archives of microbiology 163 (1995), S. 322-328 
    Details
    ISSN: 1432-072X
    Keywords: Key words Isocitrate lyase ; n-Alkane-utilizable yeast ; Candida tropicalis ; Saccharomyces cerevisiae ; Promoters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The upstream region of the isocitrate lyase gene (UPR-ICL, 1530bp) of an n-alkane-utilizable yeast, Candida tropicalis, induced gene expression in another yeast, Saccharomyces cerevisiae, when the yeasts were grown on acetate. Surprisingly, UPR-ICL displayed the same regulatory function in the bacterium Escherichia coli when grown on acetate. We determined the interesting nucleotide sequence of UPR-ICL. The deletion analysis of UPR-ICL in both cells revealed the presence of two distinct promoters: one was localized at –394 to –379 and regulated gene expression in S. cerevisiae; the other was located near the initiation codon and regulated gene expression in E. coli. The two promoter sequences were similar, but not identical to regulatory elements that have been previously reported in S. cerevisiae and E. coli, respectively. Accordingly, the possibility of novel regulatory mechanisms could not be excluded. This is an interesting example of the presence of distinct cis-acting regulatory elements responsible for the induction of gene expression in one gene by acetate in both S. cerevisiae and E. coli. Preservation of such promoters through evolution is also discussed.
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    Facts, myths and legends on the prime industrial microorganism (1995)
    Springer
    Journal of industrial microbiology and biotechnology 14 (1995), S. 514-522 
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    ISSN: 1476-5535
    Keywords: Saccharomyces cerevisiae ; Molecular taxonomy ; Classification ; Alcoholic fermentation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Archaic speculations and firmly established legends regarding the origin of the yeastSaccharomyces cerevisiae and related species are revisited in light of past and recent ecological evidence pointing to a strict association with artificial, man-made environments such as wineries and fermentation plants. The nomenclature within this industrially important group is also discussed in view of the modifications imposed from application of molecular techniques to classification.
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    Genetic and biochemical analyses of yeast RNase MRP (1995)
    Springer
    Molecular biology reports 22 (1995), S. 75-79 
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    ISSN: 1573-4978
    Keywords: ribosome synthesis ; RNA processing ; RNase MRP ; rRNA ; Saccharomyces cerevisiae ; snoRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract RNase MRP cleaves the yeast pre-rRNA at a site in internal transcribed spacer 1 (ITS1) and this cleavage can be reproducedin vitro by the highly purified enzyme. Two protein components (Pop1p and Pop2p) have been identified which are common to yeast RNase MRP and RNase P. Moreover, purified RNase P can also cleave the pre-rRNA substratein vitro, underlining the similarities between these particles. Genetic evidence suggests that RNase MRP functionally interacts with the snoRNPs which are required for other pre-rRNA processing reactions.
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    The yeast,Saccharomyces cerevisiae, RNase P/MRP ribonucleoprotein endoribonuclease family (1995)
    Springer
    Molecular biology reports 22 (1995), S. 87-93 
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    ISSN: 1573-4978
    Keywords: ribonucleoprotein endoribonuclease ; RNase MRP ; RNase P ; Saccharomyces cerevisiae ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ribonuclease P (RNase P) is a ribonucleoprotein responsible for the endonucleolytic cleavage of the 5′-termini of tRNAs. Ribonuclease MRP (RNase MRP) is a ribonucleoprotein that has the ability to cleave both mitochondrial RNA primers presumed to be involved in mitochondrial DNA replication and rRNA precursors for the production of mature rRNAs. Several lines of evidence suggest that these two ribonucleoproteins are related to each other, both functionally and evolutionarily. Both of these enzymes have activity in the nucleus and mitochondria. Each cleave their RNA substrates in a divalent cation dependent manner to generate 5′-phosphate and 3′-OH termini. In addition, the RNA subunits of both complexes can be folded into a similar secondary structure. Each can be immunoprecipitated from mammalian cells with Th antibodies. In yeast, both have been found to share at least one common protein. This review will discuss some of the recent advances in our understanding of the structure, function and evolutionary relationship of these two enzymes in the yeast,Saccharomyces cerevisiae.
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    The two-hybrid: anin vivo protein-protein interaction assay (1995)
    Springer
    Molecular biology reports 21 (1995), S. 119-127 
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    ISSN: 1573-4978
    Keywords: β-galactosidase ; fusion protein ; protein-protein interaction ; Saccharomyces cerevisiae ; transcriptional activation ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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    URL: http://dx.doi.org/10.1007/BF00986502
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    Calcitonin gene-related peptide and substance P in the pharynx and lung of the bullfrog, Rana catesbeiana (1995)
    Springer
    Cell & tissue research 279 (1995), S. 115-121 
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    ISSN: 1432-0878
    Keywords: Pharynx ; Lung ; Calcitonin gene-related peptide ; Substance P ; Coexistence ; Immunohistochemistry ; Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Indirect double immunofluorescence labelling in the pharynx and lung of the bullfrog, Rana catesbeiana, demonstrated the occurrence, distribution, and coexistence of two neuropeptides. In the pharynx, immunoreactive calcitonin gene-related peptide (CGRP) and substance P (SP) were localized in nerve fibers distributed within and just beneath the ciliated epithelium. In the lung, CGRP and SP were localized in nerve fibers in five principal locations: 1) within the smooth muscle layer in the interfaveolar septa; 2) in the luminal thickened edges of the septa; 3) around the pulmonary vasculature; 4) within, and 5) under the ciliated epithelium. Within the smooth muscle layer in the septa, luminal thickened septa, and around blood vessels, almost all fibers showed coexistence of CGRP and SP. Within and just beneath the ciliated epithelium in the thickened septa, all fibers showed coexistence of CGRP and SP. No immunoreactivity for vasoactive intestinal polypeptide, neuropeptide Y, galanin, somatostatin, FMRFamide, and leucine-and methionine-enkephalins was detected in the nerve fibers within the larynx and the lung. Together with our previous data, the present findings suggest that peptidergic mechanisms are involved in the regulation of amphibian respiratory systems throughout their life.
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    Distribution of immunoreactive neuropeptides in the pancreas of the bullfrog, Rana catesbeiana, demonstrated by immunofluorescence (1995)
    Springer
    Cell & tissue research 280 (1995), S. 307-311 
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    ISSN: 1432-0878
    Keywords: Key words: Pancreas ; Neuropeptides ; Immunohistochemistry ; Coexistence ; Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Indirect double immunofluorescence labelling for eight neuropeptides in the pancreas of the bullfrog, Rana catesbeiana, demonstrated the occurrence, distribution, and coexistence of certain neuropeptides in the exocrine and endocrine pancreas. Immunoreactivity of substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), FMRFamide (FMRF), and galanin (GAL) was localized in nerve fibers distributed between the acini and around the duct system and vasculature of the exocrine pancreas. In these regions, CGRP-immunoreactive fibers were more numerous than those containing the other five peptides. Almost all SP fibers showed coexistence of SP with CGRP, and about one third of fibers also showed coexistence of SP with VIP, NPY, FMRF, and GAL. In the endocrine pancreas, SP, CGRP, VIP, and GAL were recognized in the nerve fibers around and within the islets of Langerhans, and VIP and GAL fibers were more numerous than SP and CGRP fibers. All CGRP fibers, and about half of the VIP and GAL fibers were immunoreactive for SP. NPY- and FMRF-immunoreactive cells were found at the periphery of the islets. These findings suggest that the exocrine and endocrine pancreatic functions of the bullfrog are under the control of peptidergic innervation.
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    Ultrastructural and immunohistochemical studies of microfibril-associated components in the posterior chamber of the eye (1995)
    Springer
    Cell & tissue research 279 (1995), S. 303-313 
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    ISSN: 1432-0878
    Keywords: Microfibrils ; Ciliary zonule ; Heparan sulfate proteoglycan ; Fibrillin ; Freeze substitution ; Glycol methacrylate ; Immunohistochemistry ; Mouse (C57BL/6J) ; Chicken (White Leghorn)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Connective tissue microfibrils were observed in tissues prepared with methods believed to minimize the loss of tissue components. The eyes of C57BL/6J mice were fixed with glutaraldehyde followed by either freeze substitution, or embedding in glycol methacrylate, a water-miscible embedding medium, after limited or no dehydration. In these preparations, microfibrils were present within sheet-like layers observed in the posterior chamber of the eye. The material enclosing the microfibrils that formed the layer was also preserved, at least partially, by fixation of the tissue with uranyl acetate or potassium permanganate (KMnO4) as observed in the chick eye. This microfibril-associated material was found to be composed of heparan sulfate proteoglycan (HSPG) as shown by positive immunostaining for HSPG, as well as by identification of 4.5 nm-wide HSPG double tracks as its major constituent. When a considerable amount of this material was lost in KMnO4-fixed tissues, the remaining portion was preserved in the form of clusters of about 50 nm in width which were periodically adhered along the length of microfibrils. At the center of each cluster, a minute dark particulate structure was present. It was composed of an approximately 10 nm-wide polygonal assembly of 3.5 nm-wide ring-like structures, and was, in unfixed chick eyes, positively immunostained for fibrillin. The periodicity of HSPG clusters, and of fibrillin, along the length of immunostained microfibrils was similar, ranging from 45 nm to 65 nm. These observations indicate that fibrillin is periodically associated at the surface of “classical” microfibrils, and it may mediate the association of large amounts of HSPG to microfibrils.
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    Aromatase-immunoreactive cells are present in mouse brain areas that are known to express high levels of aromatase activity (1995)
    Springer
    Cell & tissue research 280 (1995), S. 561-574 
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    ISSN: 1432-0878
    Keywords: Aromatase ; Reproduction ; Preoptic area ; Hypothalamus ; Limbic system ; Immunohistochemistry ; Mouse (Jackson/C57)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The transformation of testosterone into estradiol in the brain plays a key role in several behavioral and physiological processes, but it has been so far impossible to localize precisely the cells of the mammalian brain containing the relevant enzyme, viz., aromatase. We have recently established an immunohistochemical technique that allows the visualization of aromatase-immunoreactive cells in the quail brain. In this species, a marked increase in the optical density of aromatase-immunoreactive cells is observed in subjects that have been treated with the aromatase inhibitor, R76713 or racemic Vorozole. This increased immunoreactivity, associated with a total blockade of aromatase activity, has been used as a tool in the present study in which the distribution of aromatase-immunoreactive material has been reassessed in the brain of mice pretreated with R76713. As expected, the aromatase inhibitor increases the density of the immunoreactive signal in mice. Strongly immunoreactive cells are found in the lateral septal region, the bed nucleus of the stria terminalis, the central amygdala, and the dorso-lateral hypothalamus. A less dense signal is also present in the medial preoptic area, the nucleus accumbens, several hypothalamic nuclei (e.g., paraventricular and ventromedial nuclei), all divisions of the amygdala, and several regions of the cortex, especially the cortex piriformis. These data demonstrate that, contrary to previous claims, aromatase-immunoreactive cells are present in all brain regions that have been shown previously to contain high aromatase activity.
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    Immunocytochemical localization of secretory acetylcholinesterase of the parasitic nematode Nippostrongylus brasiliensis (1995)
    Springer
    Cell & tissue research 280 (1995), S. 59-64 
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    ISSN: 1432-0878
    Keywords: Acetylcholinesterase ; Immunohistochemistry ; Immunoglobulin ; Nippostrongylus brasiliensis (Scolecida) ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Various parasitic nematodes secrete acetylcholinesterase (AChE). In this study, the localization of AChE in the nematode Nippostrongylus brasiliensis and the secretory forms of AChE in culture fluid were examined. A thiocholine method revealed that AChE activity was localized in the subventral glands, which have a secretory and excretory function via a duct connected to the excretory pore. By electron microscopy, AChE activity was found mainly in the matrix of secretory granules, and sometimes in the Golgi apparatus in the subventral gland cells. These results show that nematode AChE is produced and stored in the subventral glands. Monoclonal antibodies against AChE of human erythrocytes or electric rays also bound to the nematode subventral gland, suggesting immuno-cross-reactivity of AChE among these species. When AChE activity in the nematode excretory-secretory product was examined by SDS polyacrylamide gel electrophoresis combined with the thiocholine method, intense activity was demonstrated as a single band at 74kDa. Immunoblot analysis showed specific recognition of this molecule by IgE and IgG1 antibodies, but not by IgG2a antibody, in nematode-infected rat sera. These results indicate that the nematode AChE molecule produced in and secreted from the subventral glands is antigenic for the production of IgE/IgG1 in host animals.
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    Immune-complex trapping in the splenic ellipsoids of rainbow trout (Oncorhynchus mykiss) (1995)
    Springer
    Cell & tissue research 282 (1995), S. 41-48 
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    ISSN: 1432-0878
    Keywords: Key words: Ellipsoids ; Spleen ; Immune complexes ; Immunohistochemistry ; Oncorhynchus mykiss (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Rainbow trout (Oncorhynchus mykiss), immunised with horseradish peroxidase, were given horseradish peroxidase intravenously, and the trapping of antigen in the spleen was followed 1, 24, and 48 h after injection. After 1 h, the localisation of horseradish peroxidase indicated that the antigen had been extensively trapped in the walls of the splenic ellipsoids. The colocalisation of horseradish peroxidase with rainbow trout immunoglobulin M and complement factor 3 was shown with a double immunofluorescence technique and suggested that horseradish peroxidase was trapped in the form of immune complexes. After 24 and 48 h, very little horseradish peroxidase was detected in the ellipsoids, and horseradish peroxidase was mainly found in association with large cells with prominent cytoplasmic extensions. In nonimmunised fish given horseradish peroxidase intravenously, antigen was not detected in ellipsoids. Thus, the observed difference between immunised and nonimmunised trout suggests a specific role for the splenic ellipsoids in rapid immune-complex trapping and invites speculation on its significance in a secondary immune response.
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    RT97: a marker for capsaicin-insensitive sensory endings in the rat skin (1995)
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    Cell & tissue research 282 (1995), S. 155-161 
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    ISSN: 1432-0878
    Keywords: Neurofilament ; Primary afferent fibres ; Skin ; Capsaicin ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The mouse monoclonal antibody RT97, which recognises the 200-kDa neurofilament subunit in its phosphorylated form, selectively labels the somata of sensory A-fibres (large light cells) in the dorsal root ganglion of the rat. We have tested the hypothesis that this antibody also visualises large diameter sensory fibres and their end structures in peripheral tissue, in particular in the skin. RT97 immunoreactivity is found in endings that are known to be served by myelinated afferent fibres, including Meissner-like endings, Merkel discs, hair follicle receptors, Pacinian corpuscles and free nerve endings. RT97 immunoreactivity has not, however, been observed in endings of presumably unmyelinated sensory fibres (intraepidermal fibres immunoreactive for substance P and calcitonin gene-related peptide) or in sympathetic fibres innervating sweat glands and blood vessels. In addition, neither systemic (100–150 mg/kg as adults) nor perineural capsaicin pre-treatment affects RT97 immunoreactivity in the skin. The data indicate that RT97 is a useful marker in the study of the capsaicin-insensitive sensory innervation of the skin and possibly other peripheral organs.
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    Pro-enkephalin opioid peptides are abundant in porcine and bovine splenic nerves, but absent from nerves of rat, mouse, hamster, and guinea-pig spleen (1995)
    Springer
    Cell & tissue research 281 (1995), S. 143-152 
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    ISSN: 1432-0878
    Keywords: Key words: Enkephalin ; Opioid peptides ; Spleen ; Innervation ; Neuro-immunology ; Species differences ; Immunohistochemistry ; Cow ; Pig ; Guinea-pig ; Mouse ; Rat ; Dsungarian hamster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The opioidergic innervation of the mammalian spleen and possible species differences were investigated. Light-microscopic immunohistochemistry revealed that splenic nerves of bovine and porcine spleen, but not of rat, mouse, hamster and guinea-pig spleen contained proenkephalin-derived opioidergic innervation. Immunoreactivity to both prodynorphin and pro-opiomelanocortin was absent from splenic nerves. In bovine and porcine spleen, fibers immunoreactive for met-enkephalin, met-enkephalin-Arg-Phe, met-enkephalin-Arg-Gly-Leu, leu-enkephalin and peptide F formed perivascular plexus, traveled in trabecular connective tissue, and extended into the capsule. Spatial relationships with immune cells were apparent in the white and red pulp, excluding lymphoid follicles. Colocalization of enkephalin immunoreactivity with immunoreactivities for tyrosin hydroxylase, dopamin-β-hydroxylase, and neuropeptide Y, but not for substance P or calcitonin gene-related peptide were found. Our results provide evidence that opioid expression in splenic innervation is strongly species-dependent and exclusively proenkephalin-derived. Colocalization with marker enzymes of noradrenergic neurons indicates a mainly postganglionic sympathetic origin of proenkephalinergic splenic innervation. Opioidergic perivascular nerves probably control the splenic blood flow. A close interrelationship of opioidergic fibers with immune cells provides the anatomical basis for direct effects of neurally released opioids on splenic immune functions.
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    The monoclonal antibody GB 42 – a useful marker for the differentiation of myofibroblasts (1995)
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    Cell & tissue research 281 (1995), S. 231-242 
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    ISSN: 1432-0878
    Keywords: Key words: Placenta ; Stem villi ; Actin isoforms ; Myofibroblasts ; Smooth muscle cells ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The expression patterns of a variety of cytoskeletal antigens were studied in normal human tissues (placenta, umbilical cord, myometrium, colon, mammary gland, testis, skeletal muscle, myocardium) as well as in abnormal human tissues (palmar fibromatosis, fibrocystic disease of the mammary gland, mammary carcinoma). The immunohistochemical binding patterns of the monoclonal antibody GB 42 were compared to those of commercial antibodies directed against vimentin, desmin, smooth muscle myosin, pan actin, α-smooth muscle actin and γ-smooth muscle actin. Methods applied comprised immunohistochemistry on cryostat sections and paraffin sections. Immunogold immunocytochemistry was performed on Lowicryl sections. The patterns of GB 42-binding were confirmed biochemically by SDS-PAGE and Western-blotting, and quantitative amino acid analysis. Our data suggest that the monoclonal antibody GB 42 recognizes an actin isoform which is identical to, or closely related to, γ-smooth muscle actin. Unlike the commercially available antibody against γ-smooth muscle actin, GB 42 does not cross-react with α-skeletal or α-cardiac actins. The GB 42-antigen is expressed in smooth muscle cells, myoepithelial cells and in later stages of differentiation of myofibroblasts, in all the tissues investigated. Throughout the development of smooth muscle cells and myofibroblasts, the appearance of the GB 42-antigen occurs after the expression of vimentin, desmin and α-smooth muscle actin, but prior to the expression of smooth muscle myosin. GB 42 is a reliable marker for higher stages of differentiation of smooth muscle cells and myofibroblasts.
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    Immunohistochemical localization of the calcium/calmodulin-dependent protein phosphatase, calcineurin, in the mouse testis: its unique accumulation in spermatid nuclei (1995)
    Springer
    Cell & tissue research 281 (1995), S. 273-281 
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    ISSN: 1432-0878
    Keywords: Key words: Calcineurin ; Spermatogenesis ; Spermatids ; Nuclear transformation ; Immunohistochemistry ; Mouse (Jcl:ICR ; BALB/c)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunohistochemical localization of a calmodulin-dependent protein phosphatase, calcineurin, was studied in the mouse testis in relation to previous observations showing that calmodulin is unusually rich in spermatogenic stages from mid-pachytene spermatocytes to elongating spermatids. The antibodies raised against calcineurin from scallop testis reacted with subunit B, but not subunit A, of calcineurin isoforms from mouse brain and testis. Indirect immunofluorescence using these antibodies on the mouse testis revealed positive reactions only in the nuclei of round or elongating spermatids: calcineurin started to accumulate in nuclei from the acrosomal cap phase, peaked at the initial stage of nuclear elongation, and decreased thereafter. There was almost no signal in the cytoplasm; spermatogenic cells at other stages, including spermatogonia, spermatocytes, mature sperm, and other somatic cells in the seminiferous tubules were totally negative. Immuno-electron microscopy gave the same result, on the basis of measuring the density of immunogold particles. These results suggest a role for calcineurin in remodeling of the nuclear chromatin in metamorphosing spermatids.
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    Ontogeny of calbindin-D28K and calretinin in developing chick kidney (1995)
    Springer
    Cell & tissue research 279 (1995), S. 209-213 
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    ISSN: 1432-0878
    Keywords: Calcium-binding proteins ; Immunohistochemistry ; Mesonephros ; Metanephros ; Chick embryo (White leghorn)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The ontogeny of two calcium-binding proteins (calbindin-D28k and calretinin) was studied by immunohistochemical techniques in developing chick kidney. This study showed the presence of calbindin on the 5th incubation day and calretinin on the 7th incubation day in mesonephric distal and connecting tubules, and in the medial wall of the Wolffian duct. At later stages, immunostaining for these two proteins, in particular for calretinin, was also demonstrated in some metanephric proximal tubules. Glomeruli and Bowman's capsules were negative both in the mesonephros and metanephros. The presence of calretinin in the developing kidney has thus been demonstrated for the first time. The early expression of calbindin and calretinin in mesonephric distal tubules suggests their role in regulating the final excretion of calcium. The different patterns of immunoreactivity of the walls of the Wolffian duct can be correlated with their different histogenetic and histological features.
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    Morphometric analysis of atrial natriuretic peptide-containing granules in atriocytes of rats with experimental congestive heart failure (1995)
    Springer
    Cell & tissue research 279 (1995), S. 575-583 
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    ISSN: 1432-0878
    Keywords: A-V fistula ; Immunohistochemistry ; Atrial natriuretic peptides ; Congestive heart failure ; Atriocyte ; Rat (Wistar-Munich)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The morphometric characteristics of atrial natriuretic peptide-containing granules were studied in atrial myoendocrine cells of rats with aorto-caval fistula, an experimental model of congestive heart failure. A total of 6680 granules of control and aorto-caval rats were analyzed by a computerized image analysis system that evaluated the number and sectioned surface area of granules and their subcellular location. Compared with control animals, rats with congestive heart failure displayed a slight increase in the number of peripheral granules, adjacent to the sarcolemma, but not centrally located in the Golgi areas. The mean sectioned surface area of granules in rats with congestive heart failure was about 50% of that in controls, both in the right and left atria. Rats with aortocaval fistula had a higher percent of small granules and lower percent of large granules compared with controls. The data demonstrate different morphometric characteristics in atrial natriuretic peptide-containing granules in atriocytes in rats with experimental congestive heart failure; this may reflect the enhanced synthesis and release of atrial natriuretic peptide in heart failure.
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    Noradrenergic and adrenergic systems in the brain of the urodele amphibian, Pleurodeles waltlii, as revealed by immunohistochemical methods (1995)
    Springer
    Cell & tissue research 279 (1995), S. 619-627 
    Details
    ISSN: 1432-0878
    Keywords: Brain ; Noradrenaline ; Adrenaline ; Immunohistochemistry ; Pleurodeles waltlii (Urodela)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of noradrenaline and adrenaline in the brain of the urodele amphibian Pleurodeles waltlii has been studied with antibodies raised against noradrenaline and the enzymes dopamine-β-hydroxylase and phenylethanolamine-N-methyltransferase. Noradrenaline-containing cell bodies were found in the anterior preoptic area, the hypothalamic nucleus of the periventricular organ, the locus coeruleus and in the solitary tract/area postrema complex at the level of the obex. Noradrenergic fibers are widely distributed throughout the brain innervating particularly the ventrolateral forebrain, the medial amygdala, the lateral part of the posterior tubercle, the parabrachial region and the ventrolateral rhombencephalic tegmentum. Putative adrenergic cell bodies were found immediately rostral to the obex, ventral to the solitary tract. Whereas the cell bodies and their dendrites were Golgi-like stained, axons were more difficult to trace. Nevertheless, some weakly immunoreactive fibers could be traced to the basal forebrain. A comparison of these results with data previously obtained in anurans reveals not only several general features, but also some remarkable species differences.
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    URL: http://dx.doi.org/10.1007/BF00318174
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    Tyrosine hydroxylase in the cerebral ganglia of the American cockroach (Periplaneta americana L.): an immunohistochemical study (1995)
    Springer
    Cell & tissue research 282 (1995), S. 49-57 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Tyrosine hydroxylase ; Catecholamine neurons ; Invertebrate nervous system ; Immunohistochemistry ; Cerebral ganglia ; Periplaneta americana (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. We have investigated the distribution of tyrosine-hydroxylase-like immunoreactivity in the cerebral ganglia of the American cockroach, Periplaneta americana. Groups of tyrosine-hydroxylase-immunoreactive cell bodies occur in various parts of the three regions of the cerebral ganglia. In the protocerebrum, single large neurons or small groups of neurons are located in the lateral neuropil, adjacent to the calyces, and in the dorsal portion of the pars intercerebralis. Small scattered cell bodies are found in the outer layers of the optic lobe, and clusters of larger cell bodies can be found in the deutocerebrum, medial and lateral to the antennal glomeruli. Thick bundles of tyrosine-hydroxylase-positive nerve fibers traverse the neuropil in the proto- and deutocerebrum and innervate the glomerular and the nonglomerular neuropil with fine varicose terminals. Dense terminal patterns are present in the medulla and lobula of the optic lobe, the pars intercerebralis, the medial tritocerebrum, and the area surrounding the antennal glomeruli, the central body and the mushroom bodies. The pattern of tyrosine-hydroxylase-like immunoreactivity is similar to that previously described for catecholaminergic neurons, but it is distinctly different from the distribution of histaminergic and serotonergic neurons.
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    FMRFamide is endogenous to the Aplysia heart (1995)
    Springer
    Cell & tissue research 282 (1995), S. 331-341 
    Details
    ISSN: 1432-0878
    Keywords: Key words: FMRFamide ; Neuropeptide ; Immunohistochemistry ; High performance liquid chromatography ; Neurohormone ; Aplysia californica (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The presence of the molluscan neuropeptide FMRFamide was investigated in the heart of the sea hare, Aplysia californica. Immunohistochemical localization and high performance liquid chromatography (HPLC) coupled with radioimmunoassays of HPLC fractions were used to demonstrate the presence of FMRFamide and FLRFamide in the heart. FMRFamide-immunoreactive (FMRFamide-IR) nerve fibers, varicosities, and neuronal somata were observed in whole- mounts of the hearts. The atrium and atrioventricular (AV) valve regions contained significantly higher densities (P〈0.05, ANOVA) of immunoreactive varicosities compared to the ventricle. The high density of FMRFamide-IR varicosities in the atrium and the lack of sensitivity of this region to FMRFamide suggest that the atrium may be a neurohemal organ for the release of FMRFamide. The presence of FMRFamide-IR somata in the Aplysia heart suggests that peripheral neurons may play a role in modifying heart activity, independent of the central nervous system.
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    Immunohistochemistry of Grandry corpuscles in the oral mucosa of the duck bill: a light- and electron-microscopic study (1995)
    Springer
    Cell & tissue research 280 (1995), S. 253-258 
    Details
    ISSN: 1432-0878
    Keywords: Immunohistochemistry ; Substance P ; Grandry corpuscle ; Sensory nerves ; Dense-core vesicles ; Anas platyrhynchos (Aves, Anatiformes)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Grandry corpuscles in the oral mucosa of the upper bill of the duck were immunohistochemically studied using antisera against calcitonin gene-related peptide (CGRP), galanin, methionine-enkephalin, neuropeptide Y (NPY), somatostatin, substance P (SP) and vasoactive intestinal peptide (VIP). Grandry corpuscles in the lamina propria selectively showed only SP-like immunoreactivity. Herbst corpuscles distributed near Grandry corpuscles were negative to all antisera applied. Although immunoreactive products in the Grandry corpuscles were found as granules in the peripheral cytoplasm of the Grandry cell, the axon terminals and satellite cells exhibited no reactivity. In pre-embedding electron-microscopic sections, SP-like immunoreactive products visualized with 3,3′-diaminobezidine were localized in the granules of Grandry cells, but no labeling was observed in the cytoplasmic matrix or cell organelles. Electron-immunocytochemical labeling with colloidal gold by the post-embedding method clearly demonstrated that the SP antigen was localized only in the granules. It is presumed that Grandry cells have a secretory function. However, the function and the method of release of the SP contained in the observed granules remains obscure. Some CGRP-, NPY-, SP- and VIP-like-immunoreactive nerve fibers with varicosities associated with blood vessels and nerve fiber bundles of various sizes were observed in the lamina propria, but no such fibers penetrated into the intraepitherial layer. Nerve fibers positive for SP and VIP were also found in the interlobular connective tissue of the palatine glands. Some SP-positive neurons were detected in the vicinity of the palatine glands.
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    URL: http://dx.doi.org/10.1007/BF00307796
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    Distribution of peptidyl-glycine alpha-amidating monooxygenase immunoreactivity in the brain, pituitary and islet organ of the anglerfish (Lophius americanus) (1995)
    Springer
    Cell & tissue research 280 (1995), S. 159-170 
    Details
    ISSN: 1432-0878
    Keywords: Peptidyl-glycine alpha-amidating monooxygenase ; Insulin ; Glucagon ; Anglerfish peptide Y ; Neuropeptide Y ; Brain, pituitary, and islet organ ; Pancreas ; Immunohistochemistry ; Anglerfish, Lophius americanus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Peptidyl-glycine alpha-amidating monooxygenase (PAM; EC 1.14.17.3) is an enzyme that catalyzes conversion of glycine-extended peptides to alpha-amidated bioactive peptides. Two peptides that are processed at their carboxyl-termini by this enzyme are neuropeptide Y and anglerfish peptide Y, both of which possess a C-terminal glycine that is used as a substrate for amidation. Results from previous reports have demonstrated that neuropeptide Y-like and anglerfish peptide Y-like immunoreactivities are present in the brain of anglerfish (Lophius americanus). Furthermore, neuropeptide Y-like peptides, namely anglerfish peptide Y and anglerfish peptide YG (the homologues of pancreatic polypeptide) are present in the islet organ of this species. Neuropeptide Y has also been localized in the anterior, intermediated and posterior lobes of the pituitary gland in a variety of species. In order to learn more about the distribution of the enzyme responsible for alpha amidation of these peptides in the brain and pituitary and to specifically investigate the relationship of this enzyme to peptide synthesizing endocrine cells of the anglerfish islet, we performed an immunohistochemical study using several antisera generated against different peptide sequences of the enzyme. PAM antisera labeled cells in the islet organ, pituitary and brain, and fibers in the brain and pituitary gland. The PAM staining pattern in the brain was remarkably similar to the distribution of neuropeptide Y immunoreactivity reported previously. Clusters of cells adjacent to vessels in the anterior pituitary displayed punctate PAM immunoreactivity while varicose fibers were observed in the pituitary stalk and neurohypophysis. Endocrine cells of the islet organ were differentially labeled with different PAM antisera. Comparison of the staining patterns of insulin, glucagon, and anglerfish peptide Y in the islet organ to PAM immunoreactivity suggests a distribution of forms of PAM enzyme in insulin and anglerfish peptide Y-containing cells, but no overlap with glucagon-producing cells. The results also indicate that PAM immunoreactivity is widely distributed in the brain, pituitary and islet organ of anglerfish in cells that contain peptides that require presence of a C-terminal glycine for amidation.
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    Expression of the endogenous 14-kDa β-galactoside-binding lectin galectin in normal human skin (1995)
    Springer
    Cell & tissue research 280 (1995), S. 1-10 
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    ISSN: 1432-0878
    Keywords: Key words: Galectin ; β-Galactoside-binding lectin ; Human ; Skin ; Immunocytochemistry ; Immunohistochemistry ; Hybridization ; in situ ; Langerhans cell ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The localization of an endogenous 14-kDa β-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodi es. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining me thod following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the β-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.
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    Expression of the endogenous 14-kDa β-galactoside-binding lectin galectin in normal human skin (1995)
    Springer
    Cell & tissue research 280 (1995), S. 1-10 
    Details
    ISSN: 1432-0878
    Keywords: Galectin ; β-Galactoside-binding lectin ; Human ; Skin ; Immunocytochemistry ; Immunohistochemistry ; Hybridization, in situ ; Langerhans cell ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localization of an endogenous 14-kDa β-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodies. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining method following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the β-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.
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    Pro-enkephalin opioid peptides are abundant in porcine and bovine splenic nerves, but absent from nerves of rat, mouse, hamster, and guinea-pig spleen (1995)
    Springer
    Cell & tissue research 281 (1995), S. 143-152 
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    ISSN: 1432-0878
    Keywords: Enkephalin ; Opioid peptides ; Spleen ; Innervation ; Neuro-immunology ; Species differences ; Immunohistochemistry ; Cow ; Pig ; Guinea-pig ; Mouse ; Rat ; Dsungarian hamster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The opioidergic innervation of the mammalian spleen and possible species differences were investigated. Light-microscopic immunohistochemistry revealed that splenic nerves of bovine and porcine spleen, but not of rat, mouse, hamster and guinea-pig spleen contained proenkephalin-derived opioidergic innervation. Immunoreactivity to both prodynorphin and pro-opiomelanocortin was absent from splenic nerves. In bovine and porcine spleen, fibers immunoreactive for met-enkephalin, met-enkephalin-Arg-Phe, met-enkephalin-Arg-Gly-Leu, leu-enkephalin and peptide F formed perivascular plexus, traveled in trabecular connective tissue, and extended into the capsule. Spatial relationships with immune cells were apparent in the white and red pulp, excluding lymphoid follicles. Colocalization of enkephalin immunoreactivity with immunoreactivities for tyrosin hydroxylase, dopamin-β-hydroxylase, and neuropeptide Y, but not for substance P or calcitonin gene-related peptide were found. Our results provide evidence that opioid expression in splenic innervation is strongly species-dependent and exclusively proenkephalin-derived. Colocalization with marker enzymes of noradrenergic neurons indicates a mainly postganglionic sympathetic origin of proenkephalinergic splenic innervation. Opioidergic perivascular nerves probably control the splenic blood flow. A close interrelationship of opioidergic fibers with immune cells provides the anatomical basis for direct effects of neurally released opioids on splenic immune functions.
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    URL: http://dx.doi.org/10.1007/BF00307968
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    Reduced laminin immunoreactivity in the blood vessel wall of ageing rats correlates with reduced innervation in vivo and following transplantation (1995)
    Springer
    Cell & tissue research 281 (1995), S. 23-32 
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    ISSN: 1432-0878
    Keywords: Basal lamina ; Laminin ; Ageing ; Immunohistochemistry ; Confocal microscopy ; Blood vessels ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Changes in extracellular matrix composition and/or organisation, and in particular in the ratio of axonal growth-promoting components such as laminin to growth-inhibiting molecules, could contribute to the degenerative changes observed in the innervation of some peripheral tissues in old age. We have investigated this issue by evaluating laminin content or accessibility at various locations on blood vessels where we had previously studied age-related alterations in innervation density. We have employed a morphological approach, measuring laminin immunoreactivity by a densitometric application of confocal microscopy, because more conventional biochemical techniques would have been unable to distinguish specific, localized changes in laminin at sites accessible to nerves from heterogeneous changes in other areas of the vessel wall, such as the endothelial basal lamina. We found that in 24-month-old rats laminin immunoreactivity is decreased by 50% at the medial-adventitial border in association with the outer layer of smooth muscle cells, where a parallel decrease is observed in innervation density. Axonal terminals were shown to have access to laminin in this region of the blood vessel wall by double staining with laminin and a general neuronal marker. Changes in laminin immunore-activity were region-specific on the same blood vessel, thus excluding the possibility of a generalized decrease in immunoreactivity in old age. For example, in the basilar artery intensity of laminin immunoreactivity decreased in old age at the medial-adventitial border, but showed no change in endothelial cell basal lamina and in the adventitia. Moreover, we performed in oculo transplants of blood vessels displaying differences in laminin immunoreactivity and found that the density of innervation correlated with the intensity of laminin staining, thus lending further support to the hypothesis that laminin might play a role in nerve fibre atrophy in old age.
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    URL: http://dx.doi.org/10.1007/BF00307955
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    Origins of nerve fibers containing nitric oxide synthase in the rat celiac-superior mesenteric ganglion (1995)
    Springer
    Cell & tissue research 281 (1995), S. 215-221 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Nitric oxide synthase ; Immunohistochemistry ; Retrograde tracing ; Celiac-superior mesenteric ganglion ; Sensory ganglion ; Spinal cord ; Intestine ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The origin of nitric oxide synthase-containing nerve fibers in rat celiac-superior mesenteric ganglion was examined using retrograde tracing techniques combined with the immunofluorescence method. Fluoro-Gold was injected into the celiac-superior mesenteric ganglion. Neuronal cell bodies retrogradely labeled with Fluoro-Gold in the thoracic spinal cord, the dorsal root ganglia at the thoracic level, the nodose ganglion, and the intestine from the duodenum to the proximal colon were examined for nitric oxide synthase immunoreactivity. About 60% of sympathetic preganglionic neurons in the intermediolateral nucleus projecting to the celiac-superior mesenteric ganglion were immunoreactive for nitric oxide synthase, as were approximately 27% of nodose ganglion neurons and about 65% of dorsal root ganglion neurons projecting to the celiac-superior mesenteric ganglion. Neurons projecting to the celiac-superior mesenteric ganglion were found in the myenteric plexus of the small and large intestine. In the proximal colon, about 23% of such neurons were immunoreactive for nitric oxide synthase. However, in the small intestine, no immunoreactivity was found in these neurons.
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    Do vasoactive intestinal peptide (VIP)-and nitric oxide synthase-immunoreactive terminals synapse exclusively with VIP cell bodies in the submucous plexus of the guinea-pig ileum? (1995)
    Springer
    Cell & tissue research 281 (1995), S. 485-491 
    Details
    ISSN: 1432-0878
    Keywords: Nitric oxide synthase ; Vasoactive intestinal peptide ; Immunohistochemistry ; Electron microscopy ; Submucous plexus ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In the submucous plexus of the guinea-pig ileum, previous light-microscopic studies have revealed that vasoactive intestinal peptide (VIP)-immunoreactive and nitric oxide synthase (NOS)-immunoreactive terminals are found predominantly in association with VIP-immunoreactive nerve cell bodies. In this study, double-label immunohistochemistry at the light-microscopic level demonstrated co-localization of NOS-immunoreactivity and VIP-immunoreactivity in axon terminals in submucous ganglia. About 90% of nerve fibres with NOS-immunoreactivity or VIP-immunoreactivity were immunoreactive for both antigens; only about 10% of labelled varicosities contained only NOS-immunoreactivity or VIP-immunoreactivity. The VIP/NOS varicosities were more often seen in the central parts of the ganglia, close to the VIP-immunoreactive cell bodies. Ultrastructural immunocytochemistry with antibodies to VIP was used to determine if NOS/VIP terminals synapse exclusively with VIP-immunoreactive nerve cell bodies. We examined the targets of VIP-immunoreactive boutons in two submucous ganglia from different animals. Serial ultrathin sections were taken through the ganglia after they had been processed for VIP immunocytochemistry. For each cell body, the number of VIP inputs (synapses and close contacts) was determined. The number of VIP-immunoreactive synapses received by the cell bodies of submucous neurons varied from 0–4 and the number of VIP-immunoreactive close contacts varied from 3–10. There was no significant difference between VIP-immunoreactive nerve cell bodies and non-VIP nerve cell bodies in the number of VIP-immunoreactive synapses and close contacts they received. Thus, the implication from light microscopy that NOS/VIP terminals end predominantly on VIP nerve cells was not vindicated by electron microscopy.
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    Efferent projections from the retrochiasmatic area to the median eminence and to the pars nervosa of the hypophysis with special reference to the A15 dopaminergic cell group in the sheep (1995)
    Springer
    Cell & tissue research 281 (1995), S. 561-567 
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    ISSN: 1432-0878
    Keywords: Anterograde tracers ; Immunohistochemistry ; Tyrosine hydroxylase ; A15 dopaminergic group ; Retrochiasmatic area ; Prolactin secretion ; Sheep
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Anterograde tracers, viz. Phaseolus vulgaris leucoagglutinin and fluorescein dextran, were used in conjunction with tyrosine hydroxylase immunohisto-chemistry to study the projections of the A15 dopaminergic cell group towards the median eminence and pituitary in sheep. After injection of the tracers in the retrochiasmatic area, which contains the cell group A15, fibres containing anterograde tracer were observed in the internal zone of the median eminence and in the pars nervosa of the pituitary. Numerous tyrosine hydroxylase immunoreactive fibers were present in the external zone of the median eminence and in the pars intermedia and the pars nervosa of the pituitary, with characteristic patterns of organisation in each area. Most tyrosine hydroxylase-immunoreactive fibres containing fluorescein dextran were located in the pars nervosa, whereas only a few were observed in the internal zone of the median eminence. It was concluded that at least part of the dopaminergic innervation of the pars nervosa originated from the A15 group. These results provide morphological evidence for (1) the role of dopaminergic neurons of the A15 cell group in the seasonal control of prolactin secretion via the release of dopamine in the pars nervosa, and (2) putative physiological interactions between dopamine and the secretion of neurohypophysial hormones in sheep.
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    Immunohistochemical localization of spermadhesin AWN in the porcine male genital tract (1995)
    Springer
    Cell & tissue research 282 (1995), S. 175-179 
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    ISSN: 1432-0878
    Keywords: Immunohistochemistry ; Zona pellucida-binding protein ; Boar spermadhesin ; Pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Boar spermadhesin (AWN) is a 14-kDa multifunctional protein, attached to the surface of the spermatozoa and involved in sperm capacitation and zona pellucida binding. The cellular origin of AWN was previously unknown. Moreover, the region of the male genital tract in which AWN becomes attached to the surface of spermatozoa was also uncertain. By using monospecific polyclonal antibodies against AWN, the immunohistochemical distribution pattern of AWN epitopes has been investigated in tissue sections of the porcine male genital tract. Our study has revealed that AWN is synthesized in the rete testis and in the epithelium of the seminal vesicles. The latter are also the major contributors of seminal plasma AWN. In addition, immunoblotting analysis has shown that AWN is present on epididymal spermatozoa. Our results indicate that the cellular origin of spermadhesins is species-specific. The attachment of AWN to epididymal spermatozoa is probably important in developing the capacity for fertilization.
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    URL: http://dx.doi.org/10.1007/BF00319144
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    Tissue- and cell-specific distribution of creatine kinase B: A new and highly specific monoclonal antibody for use in immunohistochemistry (1995)
    Springer
    Cell & tissue research 280 (1995), S. 435-446 
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    ISSN: 1432-0878
    Keywords: Creatine kinase ; B-subunit ; Monoclonal antibody ; Immunohistochemistry ; Immuno-electron microscopy ; Western blot ; Mouse (C57BL/6) ; Rabbit (New Zealand White)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A synthetic 17-mer peptide corresponding to an unique sequence in the amino-terminal region of human creatine kinase B was used to raise a new and highly B-subunit-specific monoclonal antibody, CK-BYK/21E10. We show here that the monoclonal antibody is suitable for immunohistochemistry of unfixed frozen sections as well as formaldehyde- or Bouin-fixed, paraffin-embedded sections of human, rabbit, and mouse tissues. Moreover, in the study of cell- and tissue-specific distribution patterns, parallel Western blot analysis and immunoelectron microscopy is possible using this antibody. Our analyses demonstrate that creatine kinase B expression is restricted to a specific subset of cell types in various tissues. In brain, the B-subunit was found only in neurocytes, but not in glia cells. High expression was also observed in inner segments of photoreceptor cells and the outer plexiform layer of the retina, in the parietal cells of the stomach and in gut enterocytes, gallbladder and epithelial cells of the urogenital system. The possible roles of the creatine kinase/phosphocreatine-ATP system in these tissues are discussed.
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    Expression of neural cell adhesion molecules, NCAMs, and their polysialylated forms, PSA-NCAMs, in the developing rat pituitary gland (1995)
    Springer
    Cell & tissue research 280 (1995), S. 463-472 
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    ISSN: 1432-0878
    Keywords: NCAM ; PSA-NCAM ; Pituitary ; Development, ontogenetic ; Immunohistochemistry ; Rat(Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Neural cell adhesion molecules (NCAMs) can undergo post-translational modifications, such as the addition of polysialic acid chains, thus generating PSANCAMs, which are expressed mainly during development. Since polysialylation considerably modifies NCAM adhesivity, expression of NCAMs and PSANCAMs has been investigated in the developing hypophysis by immunohistochemistry. At embryonic day 13 (E13), an antibody against NCAM outlined all cellular profiles in the entire Rathke's pouch; this labelling persisted until adulthood. NCAM expression increased in all lobes during development and concerned all pituitary cell types. In contrast, at E13, PSA-NCAMs were only detected in the neural lobe, solely constituted of pituicytes at this stage, and the tuberal lobe, the only lobe expressing hormonal mRNA at the same stage. PSA-NCAMs expression increased in the neural lobe at E17 with the arrival of the neurosecretory fibres and persisted into adulthood. In the anterior lobe, PSA-NCAMs appeared at E15 where their distribution was similar to that of the differentiating corticotrophic cells; at subsequent stages, their expression extended to the whole anterior lobe. Only two cell types, corticotrophic and somatotrophic cells, remained labelled in the adult gland. In the intermediate lobe, melanotrophic cells never expressed PSA-NCAMs but these were expressed on folliculo-stellate cells at birth, preceding the onset of innervation. These results suggest that NCAMs and PSA-NCAMs play a role in pituitary histogenesis, cell differentiation and neurointermediate lobe innervation.
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    Changes in expression of two endogenous β-galactoside-binding isolectins in the dermis of chick embryonic skin during development in ovo and in vitro (1995)
    Springer
    Cell & tissue research 279 (1995), S. 3-12 
    Details
    ISSN: 1432-0878
    Keywords: β-Galactoside-binding lectin ; Dermis ; Skin ; Chick embryo ; Immunohistochemistry ; Keratinization ; Mucous metaplasia ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In order to elucidate the roles of metal-independent animal lectins, we systematically investigated changes in expression of 2 kinds of β-galactoside-binding isolectins (MW 14 and 16 kDa) in the dermis of chick embryonic tarsometatarsal skin during the course of development. These lectins were immunohistochemically located at different stages of development both in ovo and in vitro by light and electron microscopy. Light-microscopic observation showed that while positive staining for the 14-kDa lectin was weak at days 8 and 10 it became intense after day 13. In contrast, staining for the 16-kDa lectin was intense at days 8, 10, and 13, but it became weak after day 17 when keratinization of the epidermis was completed. Immuno-electron-microscopic observation revealed that both the 14 and 16-kDa lectins were located on the basement membrane, in the extracellular matrix, and in both the cytoplasm and the nucleus of dermal fibroblasts. Distribution of the 2 isolectins was also examined in cultured skin explants in vitro. The results were almost the same as those obtained in ovo when the skin explant was keratinized in the presence of hydrocortisone. However, in the skin explant where keratinization was prevented and mucous metaplasia was induced by the addition of vitamin A, the distribution of the 14-kDa lectin in the epidermis was significantly affected. These results indicate that (1) the expression of the 2 isolectins is differently regulated in both the dermis and epidermis, (2) the 16-kDa lectin is involved in the early stage of the formation of the dermis and the basement membrane and is replaced by the 14-kDa lectin as keratinization of the epidermis occurs, and (3) the expression of the 2 isolectins in the dermis is not significantly affected by the induction of mucous metaplasia, in contrast to their drastic changes in the epidermis.
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    Expression of neuron-specific enolase in the pineal organ of the domestic fowl during post-hatching development (1995)
    Springer
    Cell & tissue research 279 (1995), S. 25-36 
    Details
    ISSN: 1432-0878
    Keywords: Pineal organ ; Neuron-specific enolase ; Immunohistochemistry ; Three-dimensional reconstruction ; Post-hatching development ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemistry for neuron-specific enolase (NSE) revealed that NSE is localized in both a limited number of pinealocytes and intrinsic afferent neurons in the pineal organ of the domestic fowl. Furthermore, a computer-assisted three-dimensional imaging technique allowed to clarify the reverse distributional pattern of both elements: NSE-positive pinealocytes displayed a dense distribution especially in the vesicular portion of the gland, whereas NSE-immunoreactive nerve cells were mainly found in the pineal stalk. The number of NSE-positive intrinsic neurons in the pineal organ of chickens decreased rapidly after hatching, with a concentration of these elements in the basal portion (stalk) of the pineal organ. On the other hand, immunoreactive pinealocytes increased remarkably in the end-vesicle of the organ with age, followed by a gradual expansion toward the proximal portion. Thus, the spectacular increase in NSE-positive pinealocytes and the progressive reduction of reactive neurons occurred in parallel during the course of post-hatching development. NSE-immunoreactive pinealocytes displayed morphological characteristics of bipolar elements, endowed with an apical protrusion into the pineal lumen and a short basal process at younger stages, whereas multipolar types of NSE-positive pinealocytes were predominantly found in the adult domestic fowl. These results indicate that in the pineal organ of the domestic fowl (1) the ontogenetic expansion of NSE-immunoreactive pinealocytes is paralleled by a regressive afferent innervation, (2) the NSE-positive pinealocytes transform from a bipolar (columnar) type to a multipolar type during post-hatching development, and (3) these ontogenetic changes in the NSE-immunoreactivity and morphology of pinealocytes may reflect the development of a neurosecretory-like capacity of the organ.
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    URL: http://dx.doi.org/10.1007/BF00300688
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    Localization of CD44, the hyaluronate receptor; on the plasma membrane of osteocytes and osteoclasts in rat tibiae (1995)
    Springer
    Cell & tissue research 280 (1995), S. 225-233 
    Details
    ISSN: 1432-0878
    Keywords: CD44, adhesion molecule ; Bone ; Osteoclasts ; Osteocytes ; Immunohistochemistry ; Confocal laser scanning microscopy ; Electron microscopy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract CD44 is a multifunctional adhesion molecule that binds to hyaluronic acid, type I collagen, and fibronectin. We have studied the immunohistochemical localization of CD44 in bone cells by confocal laser scanning microscopy and transmission electron microscopy in order to clarify its role in the cell-cell and/or cell-matrix interaction of bone cells. In round osteoblasts attached to bone surfaces, immunoreactivity is restricted to their cytoplasmic processes. On the other hand, osteocytes in bone matrices show intense immunoreactivity on their plasma membrane. Intense immunoreactivity for CD44 can be detected on the basolateral plasma membranes of osteoclasts. There is considerably less reactivity observed in the area of the plasma membrane that is in direct contact with bone. The pre-embedding electron-microscopical method has revealed that CD44 is mainly localized on the basolateral plasma membrane of osteoclasts. However, the ruffled border and clear zone show little immunoreactivity. A CD44-positive reaction can be detected on both plasma membranes in the contact region between osteoclasts and osteocytes. These findings suggest that: 1) cells of the osteoblast lineage express CD44 in accordance with their morphological changes from osteoblasts into osteocytes; 2) osteoclasts express CD44 on their basolateral plasma membrane; 3) CD44 in osteoclasts and osteocytes may play an important role in cell-cell and/or cell-matrix attachment via extracellular matrices.
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    URL: http://dx.doi.org/10.1007/BF00307793
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    Immunophenotypic evidence for distinct populations of microglia in the rat hypothalamo-neurohypophysial system (1995)
    Springer
    Cell & tissue research 280 (1995), S. 665-673 
    Details
    ISSN: 1432-0878
    Keywords: Microglia ; Hypothalamo-neurohypophysial system ; Antigen-presenting cells ; Blood-brain barrier ; Phagocytosis ; Immunohistochemistry ; Rat (Long Evans)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The morphology, distribution and immunophenotype of microglia throughout the adult rat hypothalamo-neurohypophysial system was examined. Four macrophage-associated antibodies (OX-42, F4/80, ED1 and ED2) were used; the expression of major histocompatibility complex antigens was investigated by use of antibodies against OX-6, OX-17 (MHC class II) and OX-18 (MHC class I). Three distinct types of microglia were identified. The first was located in the magnocellular nuclei; these ‘radially branched’ (‘ramified’) microglia had round cell bodies and long branched processes, and were strongly immunoreactive only for OX-42. The second was located outside the blood-brain barrier in the median eminence, pituitary stalk and neurohypophysis often close to blood vessels; these ‘compact’ microglia had irregular cell bodies and shorter processes, and were strongly labelled by OX-42 and F4/80, weakly labelled by OX-18, and generally unlabelled by ED1, ED2, OX-6 and OX-17. The third type was found in small numbers throughout the system at the surface of the neurvous tissue or around blood vessels; these ‘perivascular’ microglia were elongated cells with no branching processes, and were strongly labelled by ED1, ED2, OX-18, OX-6, OX-17 and F4/80 antibodies but showed variable OX-42 immunoreactivity. Cells in a perivascular location were heterogeneous with respect to their immunophenotype. The presence in the normal adult rat hypothalamo-neurohypophysial system of MHC class-II molecules (OX-6 and OX-17) on a sub-set of perivascular microglia suggests that these cells are capable of presenting antigen to T lymphocytes. The microglia, which lie on either side of the blood-brain barrier, are well placed to facilitate interaction between the immune and neuroendocrine systems.
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    Configuration and distribution of bovine spermatogonia (1995)
    Springer
    Cell & tissue research 279 (1995), S. 277-289 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Spermatogonia ; Protein gene product (PGP) 9.5 ; Immunohistochemistry ; Tubular whole-mounts ; Spermatogonial degeneration ; Testis ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The configuration and distribution of bovine spermatogonia, preleptotene primary spermatocytes and Sertoli cells in the basal seminiferous tubular compartment have been studied by means of whole-mount preparations, immunohistochemistry and quantitative morphology. Three types of spermatogonia (Sg) can be identified. Large A-spermatogonia are irregularly distributed in the tubular periphery. Following the period of propagation of the A-spermatogonia, an interconnected meshwork of medium-sized spermatogonia with different cytogenetic potency is observed. Although the majority of the medium-sized spermatogonia are kinetically of the I type and divide to produce small B-spermatogonia, some members of the medium-sized population are seen in a growth phase and differentiate into large A-spermatogonia. These mark the beginning of a new round of spermatocytogenesis. Only one generation of B-spermatogonia divides into preleptotene primary spermatocytes. The architectural arrangement of multiplying spermatogonia in circles or rows is primarily the result of the distribution of the Sertoli cells. Spermatogonial multiplication is not strictly coordinated with the stages of the seminiferous epithelial cycle. Spermatogonial degeneration amounts on average to 3.6% and has therefore no decisive impact on the yield of primary spermatocytes.
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    URL: http://dx.doi.org/10.1007/s004410050283
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    Ultrastructural and immunohistochemical studies of microfibril-associated components in the posterior chamber of the eye (1995)
    Springer
    Cell & tissue research 279 (1995), S. 303-313 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Microfibrils ; Ciliary zonule ; Heparan sulfate proteoglycan ; Fibrillin ; Freeze substitution ; Glycol methacrylate ; Immunohistochemistry ; Mouse (C57BL/6J) ; Chicken (White Leghorn)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Connective tissue microfibrils were observed in tissues prepared with methods believed to minimize the loss of tissue components. The eyes of C57BL/6J mice were fixed with glutaraldehyde followed by either freeze substitution, or embedding in glycol methacrylate, a water-miscible embedding medium, after limited or no dehydration. In these preparations, microfibrils were present within sheet-like layers observed in the posterior chamber of the eye. The material enclosing the microfibrils that formed the layer was also preserved, at least partially, by fixation of the tissue with uranyl acetate or potassium permanganate (KMnO4) as observed in the chick eye. This microfibril-associated material was found to be composed of heparan sulfate proteoglycan (HSPG) as shown by positive immunostaining for HSPG, as well as by identification of 4.5 nm-wide HSPG double tracks as its major constituent. When a considerable amount of this material was lost in KMnO4-fixed tissues, the remaining portion was preserved in the form of clusters of about 50 nm in width which were periodically adhered along the length of microfibrils. At the center of each cluster, a minute dark particulate structure was present. It was composed of an approximately 10 nm-wide polygonal assembly of 3.5 nm-wide ring-like structures, and was, in unfixed chick eyes, positively immunostained for fibrillin. The periodicity of HSPG clusters, and of fibrillin, along the length of immunostained microfibrils was similar, ranging from 45 nm to 65 nm. These observations indicate that fibrillin is periodically associated at the surface of “classical” microfibrils, and it may mediate the association of large amounts of HSPG to microfibrils.
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    URL: http://dx.doi.org/10.1007/s004410050285
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    Immunophenotypic evidence for distinct populations of microglia in the rat hypothalamo-neurohypophysial system (1995)
    Springer
    Cell & tissue research 280 (1995), S. 665-673 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Microglia ; Hypothalamo-neurohypophysial system ; Antigen-presenting cells ; Blood-brain barrier ; Phagocytosis ; Immunohistochemistry ; Rat (Long Evans)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The morphology, distribution and immunophenotype of microglia throughout the adult rat hypo- thalamo-neurohypophysial system was examined. Four macrophage-associated antibodies (OX-42, F4/80, ED1 and ED2) were used; the expression of major histocompatibility complex antigens was investigated by use of antibodies against OX-6, OX-17 (MHC class II) and OX-18 (MHC class I). Three distinct types of microglia were identified. The first was located in the magnocellular nuclei; these ’radially branched’ (’ramified’) microglia had round cell bodies and long branched processes, and were strongly immunoreactive only for OX-42. The second was located outside the blood-brain barrier in the median eminence, pituitary stalk and neurohypophysis often close to blood vessels; these ’compact’ microglia had irregular cell bodies and shorter processes, and were strongly labelled by OX-42 and F4/80, weakly labelled by OX-18, and generally unlabelled by ED1, ED2, OX-6 and OX-17. The third type was found in small numbers throughout the system at the surface of the nervous tissue or around blood vessels; these ’perivascular’ microglia were elongated cells with no branching processes, and were strongly labelled by ED1, ED2, OX-18, OX-6, OX-17 and F4/80 antibodies but showed variable OX-42 immunoreactivity. Cells in a perivascular location were heterogeneous with respect to their immunophenotype. The presence in the normal adult rat hypothalamo-neurohypophysial system of MHC class-II molecules (OX-6 and OX-17) on a sub-set of perivascular microglia suggests that these cells are capable of presenting antigen to T lymphocytes. The microglia, which lie on either side of the blood-brain barrier, are well placed to facilitate interaction between the immune and neuroendocrine systems.
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    Configuration and distribution of bovine spermatogonia (1995)
    Springer
    Cell & tissue research 279 (1995), S. 277-289 
    Details
    ISSN: 1432-0878
    Keywords: Spermatogonia ; Protein gene product (PGP) 9.5 ; Immunohistochemistry ; Tubular wholemounts ; Spermatogonial degeneration ; Testis ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The configuration and distribution of bovine spermatogonia, preleptotene primary spermatocytes and Sertoli cells in the basal seminiferous tubular compartment have been studied by means of whole-mount preparations, immunohistochemistry and quantitative morphology. Three types of spermatogonia (Sg) can be identified. Large A-spermatogonia are irregularly distributed in the tubular periphery. Following the period of propagation of the A-spermatogonia, an interconnected meshwork of medium-sized spermatogonia with different cytogenetic potency is observed. Although the majority of the medium-sized spermatogonia are kinetically of the I type and divide to produce small B-spermatogonia, some members of the medium-sized population are seen in a growth phase and differentiate into large A-spermatogonia. These mark the beginning of a new round of spermatocytogenesis. Only one generation of B-spermatogonia divides into preleptotene primary spermatocytes. The architectural arrangement of multiplying spermatogonia in circles or rows is primarily the result of the distribution of the Sertoli cells. Spermatogonial multiplication is not strictly coordinated with the stages of the seminiferous epithelial cycle. Spermatogonial degeneration amounts on average to 3.6% and has therefore no decisive impact on the yield of primary spermatocytes.
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    Characterization and localization of gonadotropin-releasing hormone in the brain and pituitary of the three-spined stickleback, Gasterosteus aculeatus (1995)
    Springer
    Cell & tissue research 279 (1995), S. 485-493 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Gonadotropin-releasing hormone ; Hypothalamus ; Pituitary ; pars distalis ; High-performance liquid chromatography ; Immunohistochemistry ; Radioimmunoassay ; Stickleback ; Gasterosteus aculeatus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Radioimmunoassay (RIA) studies on high-performance liquid chromatography (HPLC) fractions of brain extracts of the three-spined stickleback, Gasterosteus aculeatus, provided evidence for at least two forms of gonadotropin-releasing hormone (GnRH). One form showed chromatographic and immunological properties similar to that of synthetic salmon GnRH (sGnRH). A second, unidentified form of GnRH eluted in the same position as chicken GnRH I (cGnRH-I); however, it did not cross-react in a cGnRH-I RIA. Furthermore, it cannot be excluded that chicken GnRH II (cGnRH-II) and maybe one other unidentified form are present in the stickleback. The distribution of GnRH in the brain of breeding adult male sticklebacks was studied by use of immunohistochemistry. Two antisera against sGnRH and antisera against mGnRH and cGnRH-II were applied on cryosections and visualized using the peroxidase-antiperoxidase method. Staining patterns were similar after incubations with all four antisera. Immunoreactive fibers were found in most parts of the brain. Three distinct groups of GnRH-immunoreactive perikarya were found in the nucleus olfactoretinalis, in the nucleus anterior periventricularis, and in the nucleus lateralis tuberis. Moreover, weakly stained cells occurred in a periventricular position in the midbrain. The proximal pars distalis of the pituitary, housing the gonadotropic cells, was richly innervated by GnRH-positive fibers. In the pars intermedia and in the rostral pars distalis, immunoreactive fibers were absent.
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    The distinct gene expression of the pro-hormone convertases in the rat heart suggests potential substrates (1995)
    Springer
    Cell & tissue research 279 (1995), S. 539-549 
    Details
    ISSN: 1432-0878
    Keywords: Key words: In situ hybridization ; Immunohistochemistry ; Pro-hormone convertases ; Cardiovascular tissues ; Pro-atrial natriuretic factor ; Pro-endothelin ; Processing ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The present study examined the distribution of the pro-hormone convertases PC1, PC2, furin, PACE4 and PC5 in the rat heart. Northern blot analysis of RNA extracted from cardiac tissues showed high levels of furin and PACE4 mRNA in the atria and ventricles, while PC5 mRNA was found to be expressed at high levels in the dorsal aorta. Although undetectable by Northern blot analysis, both PC1 and PC2 mRNA were detected by in situ hybridization and immunohistochemistry in discrete regions of the intracardiac para-aortic ganglia. In situ hybridization studies also showed that furin mRNA was observed in all cardiac tissues and cells, consistent with the previously reported ubiquitous expression of this gene. PACE4 mRNA was highly abundant in both the atria and ventricular cardiomyocytes, with low to undetectable levels observed in blood vessels. Finally, PC5 transcripts were expressed in the endothelial cells lining coronary vessels and the valve leaflets of the heart. The present localization studies in the heart and cardiac blood vessels suggests potential roles for each convertase in the processing of various neuropeptides, hormones and growth factors.
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    URL: http://dx.doi.org/10.1007/s004410050313
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    Immune-complex trapping in the splenic ellipsoids of rainbow trout (Oncorhynchus mykiss) (1995)
    Springer
    Cell & tissue research 282 (1995), S. 41-48 
    Details
    ISSN: 1432-0878
    Keywords: Ellipsoids ; Spleen ; Immune complexes ; Immunohistochemistry ; Oncorhynchus mykiss (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Rainbow trout (Oncorhynchus mykiss), immunised with horseradish peroxidase, were given horseradish peroxidase intravenously, and the trapping of antigen in the spleen was followed 1, 24, and 48 h after injection. After 1 h, the localisation of horseradish peroxidase indicated that the antigen had been extensively trapped in the walls of the splenic ellipsoids. The colocalisation of horseradish peroxidase with rainbow trout immunoglobulin M and complement factor 3 was shown with a double immunofluorescence technique and suggested that horseradish peroxidase was trapped in the form of immune complexes. After 24 and 48 h, very little horseradish peroxidase was detected in the ellipsoids, and horseradish peroxidase was mainly found in association with large cells with prominent cytoplasmic extensions. In nonimmunised fish given horseradish peroxidase intravenously, antigen was not detected in ellipsoids. Thus, the observed difference between immunised and nonimmunised trout suggests a specific role for the splenic ellipsoids in rapid immune-complex trapping and invites speculation on its significance in a secondary immune response.
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    URL: http://dx.doi.org/10.1007/BF00319131
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    Immunocytochemical localization of seminal proteins in salivary and lacrimal glands of the rat (1995)
    Springer
    Cell & tissue research 280 (1995), S. 171-181 
    Details
    ISSN: 1432-0878
    Keywords: Salivary glands ; Lacrimal gland ; Male accessory sex glands ; Immunohistochemistry ; Androgen-dependent protein secretion ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Antibodies against 10 different secretory proteins from the accessory sex glands of the male rat were used for immunohistochemical studies of salivary and lacrimal glands from intact and castrated rats, at the light- and electron-microscopic levels. In the parotid gland, secretory acinar cells showed immunoreactivity with antibodies against prostatic binding protein, cystatin-related peptide and acid phosphatase (isoenzyme pI 8.0; 5.6) typical of ventral prostate, and seminal vesicle secretion VI. Western blotting analysis indicated that immunoreactivity against prostatic binding protein was attributable to a subunit, presumably C3. Acid phosphatase pI 5.6 showed a molecular weight of 66 kDa, which is at variance with the prostatic form. Immunoreactivity for secretory transglutaminase, derived from the coagulating gland, was restricted to myoepithelial and stromal cells. In castrated animals, the immunoreactivity of acinar cells was reduced to the background level, whereas stromal transglutaminase immunoreactivity was unaltered. The distribution pattern of immunoreactivity for the proteins mentioned was almost identical in the lacrimal gland. Significant differences were however observed in the immunoreactivity of the inframandibular gland, where serous glandular cells were non-immunoreactive for seminal proteins, with the exception of acid phosphatase isoenzyme pI 8.0. Granules present in the convoluted granular ducts were immunoreactive particularly for acid phosphatase (isoenzyme pI 5.6)but much less for cystatin-related peptide; immunoreactivity was reduced after castration. The straight portion of the inframandibular duct system was immunoreactive for transglutaminase, but no influence of castration was visible. The distribution of immunoreactivity for seminal proteins present in the salivary and lacrimal glands and the pronounced androgen-dependence of their expression point to functional relationships of the respective proteins at both glandular sites.
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    Immunocytochemical localization of secretory acetylcholinesterase of the parasitic nematode Nippostrongylus brasiliensis (1995)
    Springer
    Cell & tissue research 280 (1995), S. 59-64 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Acetylcholinesterase ; Immunohistochemistry ; Immunoglobulin ; Nippostrongylusbrasiliensis (Scolecida) ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Various parasitic nematodes secrete acetylcholinesterase (AChE). In this study, the localization of AChE in the nematode Nippostrongylus brasiliensis and the secretory forms of AChE in culture fluid were examined. A thiocholine method revealed that AChE activity was localized in the subventral glands, which have a secretory and excretory function via a duct connected to the excretory pore. By electron microscopy, AChE activity was found mainly in the matrix of secretory granules, and sometimes in the Golgi apparatus in the subventral gland cells. These results show that nematode AChE is produced and stored in the subventral glands. Monoclonal antibodies against AChE of human erythrocytes or electric rays also bound to the nematode subventral gland, suggesting immuno-cross-reactivity of AChE among these species. When AChE activity in the nematode excretory-secretory product was examined by SDS polyacrylamide gel electrophoresis combined with the thiocholine met hod, intense activity was demonstrated as a single band at 74 kDa. Immunoblot analysis showed specific recognition of this molecule by IgE and IgG1 antibodies, but not by IgG2a antibody, in nematode-infected rat sera. These results indicate that the nematode AChE molecule produced in and secreted from the subventral glands is antigenic for the production of IgE/IgG1 in host animals.
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    Immunocytochemical localization of seminal proteins in salivary and lacrimal glands of the rat (1995)
    Springer
    Cell & tissue research 280 (1995), S. 171-181 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Salivary glands ; Lacrimal gland ; Male accessory sex glands ; Immunohistochemistry ; Androgen-dependent protein secretion ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Antibodies against 10 different secretory proteins from the accessory sex glands of the male rat were used for immunohistochemical studies of salivary and lacrimal glands from intact and castrated rats, at the light- and electron-microscopic levels. In the parotid gland, secretory acinar cells showed immunoreactivity with antibodies against prostatic binding protein, cystatin-related peptide and acid phosphatase (isoenzyme pI 8.0; 5.6) typical of ventral prostate, and seminal vesicle secretion VI. Western blotting analysis indicated that immunoreactivity against prostatic binding protein was attributable to a subunit, presumably C3. Acid phosphatase pI 5.6 showed a molecular weight of 66 kDa, which is at variance with the prostatic form. Immunoreactivity for secretory transglutaminase, derived from the coagulating gland, was restricted to myoepithelial and stromal cells. In castrated animals, the immunoreactivity of acinar cells was reduced to the backgroun d level, whereas stromal transglutaminase immunoreactivity was unaltered. The distribution pattern of immunoreactivity for the proteins mentioned was almost identical in the lacrimal gland. Significant differences were however observed in the immunoreactivity of the inframandibular gland, where serous glandular cells were non-immunoreactive for seminal proteins, with the exception of acid phosphatase isoenzyme pI 8.0. Granules present in the convoluted granular ducts were immunoreactive particularly for acid phosphatase (isoenzyme pI 5.6) but much less for cystatin-related peptide; immunoreactivity was reduced after castration. The straight portion of the inframandibular duct system was immunoreactive for transglutaminase, but no influence of castration was visible. The distribution of immunoreactivity for seminal proteins present in the salivary and lacrimal glands and the pronounced androgen-dependence of their expression point to functional relationships of the respective proteins at both gla ndular sites.
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    Distribution of immunoreactive neuropeptides in the pancreas of the bullfrog, Rana catesbeiana, demonstrated by immunofluorescence (1995)
    Springer
    Cell & tissue research 280 (1995), S. 307-311 
    Details
    ISSN: 1432-0878
    Keywords: Pancreas ; Neuropeptides ; Immunohistochemistry ; Coexistence ; Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Indirect double immunofluorescence labelling for eight neuropeptides in the pancreas of the bullfrog, Rana catesbeiana, demonstrated the occurrence, distribution, and coexistence of certain neuropeptides in the exocrine and endocrine pancreas. Immunoreactivity of substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), FMRFamide (FMRF), and galanin (GAL) was localized in nerve fibers distributed between the acini and around the duct system and vasculature of the exocrine pancreas. In these regions, CGRP-immunoreactive fibers were more numerous than those containing the other five peptides. Almost all SP fibers showed coexistence of SP with CGRP, and about one third of fibers also showed coexistence of SP with VIP, NPY, FMRF, and GAL. In the endocrine pancreas, SP, CGRP, VIP, and GAL were recognized in the nerve fibers around and within the islets of Langerhans, and VIP and GAL fibers were more numerous than SP and CGRP fibers. All CGRP fibers, and about half of the VIP and GAL fibers were immunoreactive for SP. NPY- and FMRF-immunoreactive cells were found at the periphery of the islets. These findings suggest that the exocrine and endocrine pancreatic functions of the bullfrog are under the control of peptidergic innervation.
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    URL: http://dx.doi.org/10.1007/BF00307803
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    Immunohistochemical examination of intraspinal serotonin neurons and fibers in the chicken lumbar spinal cord and coexistence with Leu-enkephalin (1995)
    Springer
    Cell & tissue research 282 (1995), S. 387-397 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Serotonin (5-hydroxytryptamine) ; Enkephalin ; Spinal cord ; Immunohistochemistry ; Chicken (White leghorn)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Intraspinal serotonin–positive cells and fibers were examined in the chicken lumbar spinal cord following removal of descending serotonin fibers by spinal transection. Co-localization of Leu-enkephalin immunoreactivity in intraspinal serotonin cells was also examined using a double immunofluorescence labeling technique. By one or two weeks after spinal transection, virtually all supraspinal serotonin fibers were eliminated. Intraspinal serotonin cells were located ventral or ventrolateral to the central canal corresponding to laminae VII, VIII, and IX, and the anterior funiculus. Intraspinal serotonin cells sent fibers to (1) the pia mater on the ventral or ventrolateral surface of the spinal cord; (2) vessels in the spinal cord; (3) sympathetic preganglionic column of Terni; (4) other intraspinal serotonin neurons; (5) the central canal. Some 30%–50% of the intraspinal serotonin cells co-localized with Leu-enkephalin. Intraspinal serotonin fibers co-containing Leu-enkephalin were observed in the pia mater located on the most lateral surface of the spinal cord.
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    URL: http://dx.doi.org/10.1007/s004410050490
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    Oxytocin-producing and vasopressin-producing eosinophils in the mouse spleen: immunohistochemical, immuno-electron-microscopic and in situ hybridization studies (1995)
    Springer
    Cell & tissue research 281 (1995), S. 1-10 
    Details
    ISSN: 1432-0878
    Keywords: Spleen ; Oxytocin ; Vasopressin ; Immunohistochemistry ; Immuno-electron microscopy ; In situ hybridization ; Mouse (C57BL/6)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Oxytocin-like and vasopressin-like immunoreactive cells, and the cells expressing mRNAs for these peptides in the spleen of the C57BL/6 mouse were studied by immunohistochemistry, immuno-electron microscopy and in situ hybridization. Immunoreactive cells were distributed mainly in the splenic cord and marginal zone, whereas there were few in the lymphocyte-packed periarteriolar-lymphoid sheath, lymphoid follicle and germinal center. More numerous vasopressin-positive cells were seen in the splenic cord. The colocalization of oxytocin-like and vasopressin-like immunoreactivity in the same cells was identified by the investigation of mirror sections. By the pre-embedding immuno-electron-microscopic method using antisera against oxytocin and vasopressin, immunopositive reaction products were localized in the matrix around the specific granules, small clear vesicles and mitochondrial membrane of the eosinophils. No immunoreactivity to these peptides was found within the specific granules of the eosinophils. In situ hybridization with synthetic oligonucleotide probes labeled with 32P revealed the presence of mRNAs for oxytocin and vasopressin in the cells of the spleen, the distribution of the mRNAs for these peptides being the same as that of immunopositive cells. These observations suggest that eosinophils synthesize both oxytocin and vasopressin and store them in the matrix. Possible differences in the mechanism of synthesis and storage of these peptides between peripheral eosinophils and hypothalamic neurons are discussed.
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    URL: http://dx.doi.org/10.1007/BF00307953
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    Distribution of peptidyl-glycine alpha-amidating monooxygenase immunoreactivity in the brain, pituitary and islet organ of the anglerfish (Lophius americanus) (1995)
    Springer
    Cell & tissue research 280 (1995), S. 159-170 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Peptidyl-glycine alpha-amidating monooxygenase ; Insulin ; Glucagon ; Anglerfish peptide Y ; Neuropeptide Y ; Brain ; pituitary ; and islet organ ; Pancreas ; Immunohistochemistry ; Anglerfish ; Lophiusamericanus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Peptidyl-glycine alpha-amidating monooxygenase (PAM; EC 1.14.17.3) is an enzyme that catalyzes conversion of glycine-extended peptides to alpha-amidated bioactive peptides. Two peptides that are processed at their carboxyl-termini by this enzyme are neuropeptide Y and anglerfish peptide Y, both of which possess a C-terminal glycine that is used as a substrate for amidation. Results from previous reports have demonstrated that neuropeptide Y-like and anglerfish peptide Y-like immunoreactivities are present in the brain of anglerfish (Lophius americanus). Furthermore, neuropeptide Y-like peptides, namely anglerfish peptide Y and anglerfish peptide YG (the homologues of pancreatic polypeptide) are present in the islet organ of this species. Neuropeptide Y has also been localized in the anterior, intermediate and posterior lobes of the pituitary gland in a variety of species. In order to learn more about the distribution of the enzyme responsible for alpha amidati on of these peptides in the brain and pituitary and to specifically investigate the relationship of this enzyme to peptide synthesizing endocrine cells of the anglerfish islet, we performed an immunohistochemical study using several antisera generated against different peptide sequences of the enzyme. PAM antisera labeled cells in the islet organ, pituitary and brain, and fibers in the brain and pituitary gland. The PAM staining pattern in the brain was remarkably similar to the distribution of neuropeptide Y immunoreactivity reported previously. Clusters of cells adjacent to vessels in the anterior pituitary displayed punctate PAM immunoreactivity while varicose fibers were observed in the pituitary stalk and neurohypophysis. Endocrine cells of the islet organ were differentially labeled with different PAM antisera. Comparison of the staining patterns of insulin, glucagon, and anglerfish peptide Y in the islet organ to PAM immunoreactivity suggests a distribution of forms of PAM enzyme in insulin and anglerf ish peptide Y-containing cells, but no overlap with glucagon-producing cells. The results also indicate that PAM immunoreactivity is widely distributed in the brain, pituitary and islet organ of anglerfish in cells that contain peptides that require presence of a C-terminal glycine for amidation.
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    URL: http://dx.doi.org/10.1007/BF00304521
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    Expression of neural cell adhesion molecule (N-CAM) in perisinusoidal stellate cells of the human liver (1995)
    Springer
    Cell & tissue research 283 (1995), S. 159-165 
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    ISSN: 1432-0878
    Keywords: Key words: Cell adhesion molecules ; neuronal ; Stellate cells ; Liver ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Neural cell adhesion molecule (N-CAM) is distributed in most nerve cells and some non-neural tissues. The present immunohistochemical study has revealed, for the first time, the expression of N-CAM in perisinusoidal stellate cells of the human liver. Liver specimens were stained with monoclonal antibody against human Leu19 (N-CAM) by a streptoavidin-biotin-peroxidase-complex method. Light- and electron-microscopic analyses have shown that N-CAM-positive nerve fibers are distributed in the periportal and intermediate zones of the liver lobule. Perisinusoidal stellate cells in these zones are also positive for N-CAM. N-CAM is expressed on the surface of the cell, including cytoplasmic projections. Close contact of N-CAM-positive nerve endings with N-CAM-positive stellate cells has been observed. On the other hand, stellate cells in the centrilobular zone exhibit weak or no reaction for N-CAM. Perivascular smooth muscle cells and fibroblasts in the portal area and myofibroblasts around the central veins are negative for N-CAM. The present results indicate that the perisinusoidal stellate cells in the periportal and intermediate zones of the liver lobule characteristically express N-CAM, unlike other related mesenchymal cells, and suggest that the intralobular heterogeneity of N-CAM expression by stellate cells is related to the different maturational stages of these cells.
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    URL: http://dx.doi.org/10.1007/s004410050524
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    RT97: a marker for capsaicin-insensitive sensory endings in the rat skin (1995)
    Springer
    Cell & tissue research 282 (1995), S. 155-161 
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    ISSN: 1432-0878
    Keywords: Key words: Neurofilament ; Primary afferent fibres ; Skin ; Capsaicin ; Immunohistochemistry ; Rat (Wis-tar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The mouse monoclonal antibody RT97, which recognises the 200-kDa neurofilament subunit in its phosphorylated form, selectively labels the somata of sensory A-fibres (large light cells) in the dorsal root ganglion of the rat. We have tested the hypothesis that this antibody also visualises large diameter sensory fibres and their end structures in peripheral tissue, in particular in the skin. RT97 immunoreactivity is found in endings that are known to be served by myelinated afferent fibres, including Meissner-like endings, Merkel discs, hair follicle receptors, Pacinian corpuscles and free nerve endings. RT97 immunoreactivity has not, however, been observed in endings of presumably unmyelinated sensory fibres (intraepidermal fibres immunoreactive for substance P and calcitonin gene-related peptide) or in sympathetic fibres innervating sweat glands and blood vessels. In addition, neither systemic (100–150 mg/kg as adults) nor perineural capsaicin pre-treatment affects RT97 immunoreactivity in the skin. The data indicate that RT97 is a useful marker in the study of the capsaicin-insensitive sensory innervation of the skin and possibly other peripheral organs.
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    URL: http://dx.doi.org/10.1007/s004410050468
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    Do vasoactive intestinal peptide (VIP)- and nitric oxide synthase-immunoreactive terminals synapse exclusively with VIP cell bodies in the submucous plexus of the guinea-pig ileum? (1995)
    Springer
    Cell & tissue research 281 (1995), S. 485-491 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Nitric oxide synthase ; Vasoactive intestinal peptide ; Immunohistochemistry ; Electron microscopy ; Submucous plexus ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In the submucous plexus of the guinea-pig ileum, previous light-microscopic studies have revealed that vasoactive intestinal peptide (VIP)-immunoreactive and nitric oxide synthase (NOS)-immunoreactive terminals are found predominantly in association with VIP-immunoreactive nerve cell bodies. In this study, double-label immunohistochemistry at the light-microscopic level demonstrated co-localization of NOS-immunoreactivity and VIP-immunoreactivity in axon terminals in submucous ganglia. About 90% of nerve fibres with NOS-immunoreactivity or VIP-immunoreactivity were immunoreactive for both antigens; only about 10% of labelled varicosities contained only NOS-immunoreactivity or VIP-immunoreactivity. The VIP/NOS varicosities were more often seen in the central parts of the ganglia, close to the VIP-immunoreactive cell bodies. Ultrastructural immunocytochemistry with antibodies to VIP was used to determine if NOS/VIP terminals synapse exclusively with VIP-immunoreactive nerve cell bodies. We examined the targets of VIP-immunoreactive boutons in two submucous ganglia from different animals. Serial ultrathin sections were taken through the ganglia after they had been processed for VIP immunocytochemistry. For each cell body, the number of VIP inputs (synapses and close contacts) was determined. The number of VIP-immunoreactive synapses received by the cell bodies of submucous neurons varied from 0–4 and the number of VIP-immunoreactive close contacts varied from 3–10. There was no significant difference between VIP-immunoreactive nerve cell bodies and non-VIP nerve cell bodies in the number of VIP-immunoreactive synapses and close contacts they received. Thus, the implication from light microscopy that NOS/VIP terminals end predominantly on VIP nerve cells was not vindicated by electron microscopy.
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    URL: http://dx.doi.org/10.1007/s004410050443
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    Localization of CD44, the hyaluronate receptor, on the plasma membrane of osteocytes and osteoclasts in rat tibiae (1995)
    Springer
    Cell & tissue research 280 (1995), S. 225-233 
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    ISSN: 1432-0878
    Keywords: Key words: CD44 ; adhesion molecule ; Bone ; Osteoclasts ; Osteocytes ; Immunohistochemistry ; Confocal laser scanning microscopy ; Electron microscopy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. CD44 is a multifunctional adhesion molecule that binds to hyaluronic acid, type I collagen, and fibronectin. We have studied the immunohistochemical localization of CD44 in bone cells by confocal laser scanning microscopy and transmission electron microscopy in order to clarify its role in the cell-cell and/or cell-matrix interaction of bone cells. In round osteoblasts attached to bone surfaces, immunoreactivity is restricted to their cytoplasmic processes. On the other hand, osteocytes in bone matrices show intense immunoreactivity on their plasma membrane. Intense immunoreactivity for CD44 can be detected on the basolateral plasma membranes of osteoclasts. There is considerably less reactivity observed in the area of the plasma membrane that is in direct contact with bone. The pre-embedding electron-microscopical method has revealed that CD44 is mainly localized on the basolateral plasma membrane of osteoclasts. However, the ruffled border and clear zone show little immunoreactivity. A CD44-positive reaction can be detected on both plasma membranes in the contact region between osteoclasts and osteocytes. These findings suggest that: 1) cells of the osteoblast lineage express CD44 in accordance with their morphological changes from osteoblasts into osteocytes; 2) osteoclasts express CD44 on their basolateral plasma membrane; 3) CD44 in osteoclasts and osteocytes may play an important role in cell-cell and/or cell-matrix attachment via extracellular matrices.
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    URL: http://dx.doi.org/10.1007/BF00307793
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    Chemical codes of sensory neurons innervating the guinea-pig adrenal gland (1995)
    Springer
    Cell & tissue research 279 (1995), S. 169-181 
    Details
    ISSN: 1432-0878
    Keywords: Adrenal gland ; Dorsal root ganglia ; Immunohistochemistry ; Neurofilament ; Neuronal tracing ; Neuropeptides ; Nitric oxide synthase ; Substance P ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Retrograde neuronal tracing in combination with double-labelling immunofluorescence was applied to distinguish the chemical coding of guinea-pig primary sensory neurons projecting to the adrenal medulla and cortex. Seven subpopulations of retrogradely traced neurons were identified in thoracic spinal ganglia T1-L1. Five subpopulations contained immunolabelling either for calcitonin gene-related peptide (CGRP) alone (I), or for CGRP, together with substance (P (II), substance P/dynorphin (III), substance P/cholecystokinin (IV), and substance P/nitric oxide synthase (V), respectively. Two additional subpopulations of retrogradely traced neurons were distinct from these groups: neurofilament-immunoreactive neurons (VI), and cell bodies that were nonreactive to either of the antisera applied (VII). Nerve fibres in the adrenal medulla and cortex were equipped with the mediator combinations I, II, IV and VI. An additional meshwork of fibres solely labelled for nitric oxide synthase was visible in the medulla. Medullary as well as cortical fibres along endocrine tissue apparently lacked the chemical code V, while in the external cortex some fibres exhibited code III. Some intramedullary neuronal cell bodies revealed immunostaining for nitric oxide synthase, CGRP or substance P, providing an additional intrinsic adrenal innervation. Perikarya, immunolabelled for nitric oxide synthase, however, were too few to match with the large number of intramedullary nitric oxide synthase-immunoreactive fibres. A non-sensory participation is also supposed for the particularly dense intramedullary network of solely neurofilament-immunoreactive nerve fibres. The findings give evidence for a differential sensory innervation of the guineapig adrenal cortex and medulla. Specific sensory neuron subpopulations suggest that nervous control of adrenal functions is more complex than hitherto believed.
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    URL: http://dx.doi.org/10.1007/BF00300702
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    Areas of asbestoid (amianthoid) fibers in human thyroid cartilage characterized by immunolocalization of collagen types I, II, IX, XI and X (1995)
    Springer
    Cell & tissue research 280 (1995), S. 349-354 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Thyroid cartilage ; Immunohistochemistry ; Asbestoid fibers ; Amianthoid fibers ; Collagens ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The distribution of type I, II, IX, XI and X collagens in and close to areas of asbestoid (amianthoid) fibers in thyroid cartilages of various ages was investigated in this study. Asbestoid fibers were first detected in thyroid cartilage from a 3-year-old male child. Areas of asbestoid fibers functionally appear to serve as guide rails for vascularization of thyroid cartilage. Alcian blue staining in the presence of 0.3 M MgCl2 revealed a loss of glycosaminoglycans in areas of asbestoid fibers. In addition, the fibers reacted positively with antibodies against collagen types II, IX and XI, but showed no staining with antibodies to collagen types I and X. Territorial matrix of adjacent chondrocytes showed the same staining pattern. In addition to staining for type II, IX and XI collagens, asbestoid fibers showed strong immunostaining for type I collagen after puberty but not for type X collagen. However, groups of chondrocytes within areas of asbestoid fibers reacted strongly with antibodies to type X collagen, suggesting that this collagen plays an important role in matrix of highly differentiated chondrocytes. The finding that these type X collagen-positive chondrocytes also revealed immunostaining for type I collagen confirms previous studies showing that hypertrophic chondrocytes can further differentiate into cells that are characterized by the synthesis of type X and I collagens.
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    Tissue- and cell-specific distribution of creatine kinase B: A new and highly specific monoclonal antibody for use in immunohistochemistry (1995)
    Springer
    Cell & tissue research 280 (1995), S. 435-446 
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    ISSN: 1432-0878
    Keywords: Key words: Creatine kinase ; B-subunit ; Monoclonal antibody ; Immunohistochemistry ; Immuno-electron microscopy ; Western blot ; Mouse (C57BL/6) ; Rabbit (New Zealand White)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. A synthetic 17-mer peptide corresponding to an unique sequence in the amino-terminal region of human creatine kinase B was used to raise a new and highly B-subunit-specific monoclonal antibody, CK-BYK/21E10. We show here that the monoclonal antibody is suitable for immunohistochemistry of unfixed frozen sections as well as formaldehyde- or Bouin-fixed, paraffin-embedded sections of human, rabbit, and mouse tissues. Moreover, in the study of cell- and tissue-specific distribution patterns, parallel Western blot analysis and immuno-electron microscopy is possible using this antibody. Our analyses demonstrate that creatine kinase B expression is restricted to a specific subset of cell types in various tissues. In brain, the B-subunit was found only in neurocytes, but not in glia cells. High expression was also observed in inner segments of photoreceptor cells and the outer plexiform layer of the retina, in the parietal cells of the stomach and in gut enterocytes, gallbladder and epithelial cells of the urogenital system. The possible roles of the creatine kinase/phosphocreatine-ATP system in these tissues are discussed.
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    URL: http://dx.doi.org/10.1007/BF00307817
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    Expression of neural cell adhesion molecules, NCAMs, and their polysialylated forms, PSA-NCAMs, in the developing rat pituitary gland (1995)
    Springer
    Cell & tissue research 280 (1995), S. 463-472 
    Details
    ISSN: 1432-0878
    Keywords: Key words: NCAM ; PSA-NCAM ; Pituitary ; Development ; ontogenetic ; Immunohistochemistry ; Rat(Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Neural cell adhesion molecules (NCAMs) can undergo post-translational modifications, such as the addition of polysialic acid chains, thus generating PSA-NCAMs, which are expressed mainly during development. Since polysialylation considerably modifies NCAM adhesivity, expression of NCAMs and PSA-NCAMs has been investigated in the developing hypophysis by immunohistochemistry. At embryonic day 13 (E13), an antibody against NCAM outlined all cellular profiles in the entire Rathke’s pouch; this labelling persisted until adulthood. NCAM expression increased in all lobes during development and concerned all pituitary cell types. In contrast, at E13, PSA-NCAMs were only detected in the neural lobe, solely constituted of pituicytes at this stage, and the tuberal lobe, the only lobe expressing hormonal mRNA at the same stage. PSA-NCAMs expression increased in the neural lobe at E17 with the arrival of the neurosecretory fibres and persisted into adulthood. In the anterior lobe, PSA-NCAMs appeared at E15 where their distribution was similar to that of the differentiating corticotrophic cells; at sub- sequent stages, their expression extended to the whole anterior lobe. Only two cell types, corticotrophic and somatotrophic cells, remained labelled in the adult gland. In the intermediate lobe, melanotrophic cells never expressed PSA-NCAMs but these were expressed on folliculo-stellate cells at birth, preceding the onset of innervation. These results suggest that NCAMs and PSA-NCAMs play a role in pituitary histogenesis, cell differentiation and neurointermediate lobe innervation.
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    Aromatase-immunoreactive cells are present in mouse brain areas that are known to express high levels of aromatase activity (1995)
    Springer
    Cell & tissue research 280 (1995), S. 561-574 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Aromatase ; Reproduction ; Preoptic area ; Hypothalamus ; Limbic system ; Immunohistochemistry ; Mouse (Jackson/C57)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The transformation of testosterone into estradiol in the brain plays a key role in several behavioral and physiological processes, but it has been so far impossible to localize precisely the cells of the mammalian brain containing the relevant enzyme, viz., aromatase. We have recently established an immunohistochemical technique that allows the visualization of aromatase-immunoreactive cells in the quail brain. In this species, a marked increase in the optical density of aromatase-immunoreactive cells is observed in subjects that have been treated with the aromatase inhibitor, R76713 or racemic Vorozole. This increased immunoreactivity, associated with a total blockade of aromatase activity, has been used as a tool in the present study in which the distribution of aromatase-immunoreactive material has been reassessed in the brain of mice pretreated with R76713. As expected, the aromatase inhibitor increases the density of the immunoreactive signal in mice. Strongly immunoreactive cells are found in the lateral septal region, the bed nucleus of the stria terminalis, the central amygdala, and the dorso-lateral hypothalamus. A less dense signal is also present in the medial preoptic area, the nucleus accumbens, several hypothalamic nuclei (e.g., paraventricular and ventromedial nuclei), all divisions of the amygdala, and several regions of the cortex, especially the cortex piriformis. These data demonstrate that, contrary to previous claims, aromatase-immunoreactive cells are present in all brain regions that have been shown previously to contain high aromatase activity.
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    Reduced laminin immunoreactivity in the blood vessel wall of ageing rats correlates with reduced innervation in vivo and following transplantation (1995)
    Springer
    Cell & tissue research 281 (1995), S. 23-32 
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    ISSN: 1432-0878
    Keywords: Key words: Basal lamina ; Laminin ; Ageing ; Immunohistochemistry ; Confocal microscopy ; Blood vessels ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Changes in extracellular matrix composition and/or organisation, and in particular in the ratio of axonal growth-promoting components such as laminin to growth-inhibiting molecules, could contribute to the degenerative changes observed in the innervation of some peripheral tissues in old age. We have investigated this issue by evaluating laminin content or accessibility at various locations on blood vessels where we had pre- viously studied age-related alterations in innervation density. We have employed a morphological approach, measuring laminin immunoreactivity by a densitometric application of confocal microscopy, because more conventional biochemical techniques would have been unable to distinguish specific, localized changes in laminin at sites accessible to nerves from heterogeneous changes in other areas of the vessel wall, such as the endothelial basal lamina. We found that in 24-month-old rats laminin immunoreactivity is decreased by 50% at the medial-adventitial border in association with the outer layer of smooth muscle cells, where a parallel decrease is observed in innervation density. Axonal terminals were shown to have access to laminin in this region of the blood vessel wall by double staining with laminin and a general neuronal marker. Changes in laminin immunoreactivity were region-specific on the same blood vessel, thus excluding the possibility of a generalized decrease in immunoreactivity in old age. For example, in the basilar artery intensity of laminin immunoreactivity decreased in old age at the medial-adventitial border, but showed no change in endothelial cell basal lamina and in the adventitia. Moreover, we performed in oculo transplants of blood vessels displaying differences in laminin immunoreactivity and found that the density of innervation correlated with the intensity of laminin staining, thus lending further support to the hypothesis that laminin might play a role in nerve fibre atrophy in old age.
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    URL: http://dx.doi.org/10.1007/BF00307955
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    Characterization and localization of gonadotropin-releasing hormone in the brain and pituitary of the three-spined stickleback, Gasterosteus aculeatus (1995)
    Springer
    Cell & tissue research 279 (1995), S. 485-493 
    Details
    ISSN: 1432-0878
    Keywords: Gonadotropin-releasing hormone ; Hypothalamus ; Pituitary ; pars distalis ; High-performance liquid chromatography ; Immunohistochemistry ; Radioimmunoassay ; Stickleback, Gasterosteus aculeatus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Radioimmunoassay (RIA) studies on highperformance liquid chromatography (HPLC) fractions of brain extracts of the three-spined stickleback, Gasterosteus aculeatus, provided evidence for at least two forms of gonadotropin-releasing hormone (GnRH). One form showed chromatographic and immunological properties similar to that of synthetic salmon GnRH (sGnRH). A second, unidentified form of GnRH eluted in the same position as chicken GnRH I (cGnRH-I); however, it did not cross-react in a cGnRH-I RIA. Furthermore, it cannot be excluded that chicken GnRH II (cGnRH-II) and maybe one other unidentified form are present in the stickleback. The distribution of GnRH in the brain of breeding adult male sticklebacks was studied by use of immunohistochemistry. Two antisera against sGnRH and antisera against mGnRH and cGnRH-II were applied on cryosections and visualized using the peroxidase-antiperoxidase method. Staining patterns were similar after incubations with all four antisera. Immunoreactive fibers were found in most parts of the brain. Three distinct groups of GnRH-immunoreactive perikarya were found in the nucleus olfactoretinalis, in the nucleus anterior periventricularis, and in the nucleus lateralis tuberis. Moreover, weakly stained cells occurred in a periventricular position in the midbrain. The proximal pars distalis of the pituitary, housing the gonadotropic cells, was richly innervated by GnRH-positive fibers. In the pars intermedia and in the rostral pars distalis, immunoreactive fibers were absent.
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    URL: http://dx.doi.org/10.1007/BF00318162
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    The distinct gene expression of the pro-hormone convertases in the rat heart suggests potential substrates (1995)
    Springer
    Cell & tissue research 279 (1995), S. 539-549 
    Details
    ISSN: 1432-0878
    Keywords: In situ hybridization ; Immunohistochemistry ; Pro-hormone convertases ; Cardiovascular tissues ; Pro-atrial natriuretic factor ; Pro-endothelin ; Processing ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The present study examined the distribution of the pro-hormone convertases PC1, PC2, furin, PACE4 and PC5 in the rat heart. Northern blot analysis of RNA extracted from cardiac tissues showed high levels of furin and PACE4 mRNA in the atria and ventricles, while PC5 mRNA was found to be expressed at high levels in the dorsal aorta. Although undetectable by Northern blot analysis, both PC1 and PC2 mRNA were detected by in situ hybridization and immunohistochemistry in discrete regions of the intracardiac para-aortic ganglia. In situ hybridization studies also showed that furin mRNA was observed in all cardiac tissues and cells, consistent with the previously reported ubiquitous expression of this gene. PACE4 mRNA was highly abundant in both the atria and ventricular cardiomyocytes, with low to undetectable levels observed in blood vessels. Finally, PC5 transcripts were expressed in the endothelial cells lining coronary vessels and the valve leaflets of the heart. The present localization studies in the heart and cardiac blood vessels suggests potential roles for each convertase in the processing of various neuropeptides, hormones and growth factors.
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    URL: http://dx.doi.org/10.1007/BF00318166
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    Immunogold labelling of estradiol receptor in MCF7 cells (1995)
    Springer
    Cell & tissue research 279 (1995), S. 445-452 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Estradiol receptor ; Breast cancer cells ; Cell culture ; Ultrastructure ; Electron microscopy ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The distribution of estradiol receptor in serial sections of estradiol-deprived and estradiol-stimulated MCF7 cells was studied by using mouse monoclonal antibodies reacting with different domains of the receptor and goat-antimouse IgG/6 nm gold. In the nucleus and the cytoplasm of estradiol-deprived cells, the receptor was detected by all three monoclonals (13H2, HT 65 and MA1-310). The antibodies 13H2 and MA1-310 detected receptor associated to the microfilament bundles in the cytoplasm. Higher densities of antireceptor attachment to the nuclear areas were accompanied by a reduction in the attachment to the cytoplasm after estradiol stimulation of the cells. The results confirm earlier observations on the presence of cytoplasmic estrogen receptor in estradiol-deprived cells and support the premise of an es- tradiol-induced translocation of this ligand-dependent transcription regulator.
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    URL: http://dx.doi.org/10.1007/s004410050303
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    Morphometric analysis of atrial natriuretic peptide-containing granules in atriocytes of rats with experimental congestive heart failure (1995)
    Springer
    Cell & tissue research 279 (1995), S. 575-583 
    Details
    ISSN: 1432-0878
    Keywords: Key words: A-V fistula ; Immunohistochemistry ; Atrial natriuretic peptides ; Congestive heart failure ; Atriocyte ; Rat (Wistar-Munich)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The morphometric characteristics of atrial natriuretic peptide-containing granules were studied in atrial myoendocrine cells of rats with aorto-caval fistula, an experimental model of congestive heart failure. A total of 6680 granules of control and aorto-caval rats were analyzed by a computerized image analysis system that evaluated the number and sectioned surface area of granules and their subcellular location. Compared with control animals, rats with congestive heart failure displayed a slight increase in the number of peripheral granules, adjacent to the sarcolemma, but not centrally located in the Golgi areas. The mean sectioned surface area of granules in rats with congestive heart failure was about 50% of that in controls, both in the right and left atria. Rats with aorto-caval fistula had a higher percent of small granules and lower percent of large granules compared with controls. The data demonstrate different morphometric characteristics in atrial natriuretic peptide-containing granules in atriocytes in rats with experimental congestive heart failure; this may reflect the enhanced synthesis and release of atrial natriuretic peptide in heart failure.
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    URL: http://dx.doi.org/10.1007/s004410050316
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    Noradrenergic and adrenergic systems in the brain of the urodele amphibian, Pleurodeles waltlii, as revealed by immunohistochemical methods (1995)
    Springer
    Cell & tissue research 279 (1995), S. 619-627 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Brain ; Noradrenaline ; Adrenaline ; Immunohistochemistry ; Pleurodeles waltlii (Urodela)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The distribution of noradrenaline and adrenaline in the brain of the urodele amphibian Pleurodeles waltlii has been studied with antibodies raised against noradrenaline and the enzymes dopamine-β-hydroxylase and phenylethanolamine-N-methyltransferase. Noradrenaline-containing cell bodies were found in the anterior preoptic area, the hypothalamic nucleus of the periventricular organ, the locus coeruleus and in the solitary tract/area postrema complex at the level of the obex. Noradrenergic fibers are widely distributed throughout the brain innervating particularly the ventrolateral forebrain, the medial amygdala, the lateral part of the posterior tubercle, the parabrachial region and the ventrolateral rhombencephalic tegmentum. Putative adrenergic cell bodies were found immediately rostral to the obex, ventral to the solitary tract. Whereas the cell bodies and their dendrites were Golgi-like stained, axons were more difficult to trace. Nevertheless, some weakly immunoreactive fibers could be traced to the basal forebrain. A comparison of these results with data previously obtained in anurans reveals not only several general features, but also some remarkable species differences.
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    Immunogold labelling of estradiol receptor in MCF7 cells (1995)
    Springer
    Cell & tissue research 279 (1995), S. 445-452 
    Details
    ISSN: 1432-0878
    Keywords: Estradiol receptor ; Breast cancer cells ; Cell culture ; Ultrastructure ; Electron microscopy ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of estradiol receptor in serial sections of estradiol-deprived and estradiol-stimulated MCF7 cells was studied by using mouse monoclonal antibodies reacting with different domains of the receptor and goat-antimouse IgG/6 nm gold. In the nucleus and the cytoplasm of estradiol-deprived cells, the receptor was detected by all three monoclonals (13H2, HT 65 and MA1-310). The antibodies 13H2 and MA1-310 detected receptor associated to the microfilament bundles in the cytoplasm. Higher densities of antireceptor attachment to the nuclear areas were accompanied by a reduction in the attachment to the cytoplasm after estradiol stimulation of the cells. The results confirm earlier observations on the presence of cytoplasmic estrogen receptor in estradiol-deprived cells and support the premise of an estradiol-induced translocation of this ligand-dependent transcription regulator.
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    URL: http://dx.doi.org/10.1007/BF00318156
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    FMRFamide is endogenous to the Aplysia heart (1995)
    Springer
    Cell & tissue research 282 (1995), S. 331-341 
    Details
    ISSN: 1432-0878
    Keywords: FMRFamide ; Neuropeptide ; Immunohistochemistry ; High performance liquid chromatography ; Neurohormone ; Aplysia californica (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The presence of the molluscan neuropeptide FMRFamide was investigated in the heart of the sea hare, Aplysia californica. Immunohistochemical localization and high performance liquid chromatography (HPLC) coupled with radioimmunoassays of HPLC fractions were used to demonstrate the presence of FMRFamide and FLRFamide in the heart. FMRFamide-immunoreactive (FMRFamide-IR) nerve fibers, varicosities, and neuronal somata were observed in whole-mounts of the hearts. The atrium and atrioventricular (AV) valve regions contained significantly higher densities (P〈0.05, ANOVA) of immunoreactive varicosities compared to the ventricle. The high density of FMRF-amide-IR varicosities in the atrium and the lack of sensitivity of this region to FMRFamide suggest that the atrium may be a neurohemal organ for the release of FMRF-amide. The presence of FMRFamide-IR somata in the Aplysia heart suggests that peripheral neurons may play a role in modifying heart activity, independent of the central nervous system.
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    URL: http://dx.doi.org/10.1007/BF00319123
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    Immunohistochemical examination of intraspinal serotonin neurons and fibers in the chicken lumbar spinal cord and coexistence with Leu-enkephalin (1995)
    Springer
    Cell & tissue research 282 (1995), S. 387-397 
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    ISSN: 1432-0878
    Keywords: Serotonin (5-hydroxytryptamine) ; Enkephalin ; Spinal cord ; Immunohistochemistry ; Chicken (White leghorn)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Intraspinal serotonin-positive cells and fibers were examined in the chicken lumbar spinal cord following removal of descending serotonin fibers by spinal transection. Co-localization of Leu-enkephalin immunoreactivity in intraspinal serotonin cells was also examined using a double immunofluorescence labeling technique. By one or two weeks after spinal transection, virtually all supraspinal serotonin fibers were eliminated. Intraspinal serotonin cells were located ventral or ventrolateral to the central canal corresponding to laminae VII, VIII, and IX, and the anterior funiculus. Intraspinal serotonin cells sent fibers to (1) the pia mater on the ventral or ventrolateral surface of the spinal cord; (2) vessels in the spinal cord; (3) sympathetic preganglionic column of Terni; (4) other intraspinal serotonin neurons; (5) the central canal. Some 30%–50% of the intraspinal serotonin cells co-localized with Leu-enkephalin. Intraspinal serotonin fibers co-containing Leu-enkephalin were observed in the pia mater located on the most lateral surface of the spinal cord.
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    Areas of asbestoid (amianthoid) fibers in human thyroid cartilage characterized by immunolocalization of collagen types I, II, IX, XI and X (1995)
    Springer
    Cell & tissue research 280 (1995), S. 349-354 
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    ISSN: 1432-0878
    Keywords: Thyroid cartilage ; Immunohistochemistry ; Asbestoid fibers ; Amianthoid fibers ; Collagens ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of type I, II, IX, XI and X collagens in and close to areas of asbestoid (amianthoid) fibers in thyroid cartilages of various ages was investigated in this study. Asbestoid fibers were first detected in thyroid cartilage from a 3-year-old male child. Areas of asbestoid fibers functionally appear to serve as guide rails for vascularization of thyroid cartilage. Alcian blue staining in the presence of 0.3 M MgCl2 revealed a loss of glycosaminoglycans in areas of asbestoid fibers. In addition, the fibers reacted positively with antibodies against collagen types II, IX and XI, but showed no staining with antibodies to collagen types I and X. Territorial matrix of adjacent chondrocytes showed the same staining pattern. In addition to staining for type II, IX and XI collagens, asbestoid fibers showed strong immunostaining for type I collagen after puberty but not for type X collagen. However, groups of chondrocytes within areas of asbestoid fibers reacted strongly with antibodies to type X collagen, suggesting that this collagen plays an important role in matrix of highly differentiated chondrocytes. The finding that these type X collagen-positive chondrocytes also revealed immunostaining for type I collagen confirms previous studies showing that hypertrophic chondrocytes can further differentiate into cells that are characterized by the synthesis of type X and I collagens.
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    Efferent projections from the retrochiasmatic area to the median eminence and to the pars nervosa of the hypophysis with special reference to the A15 dopaminergic cell group in the sheep (1995)
    Springer
    Cell & tissue research 281 (1995), S. 561-567 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Anterograde tracers ; Immunohistochemistry ; Tyrosine hydroxylase ; A15 dopaminergic group ; Retrochiasmatic area ; Prolactin secretion ; Sheep
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Anterograde tracers, viz. Phaseolus vulgaris leucoagglutinin and fluorescein dextran, were used in conjunction with tyrosine hydroxylase immunohistochemistry to study the projections of the A15 dopaminergic cell group towards the median eminence and pituitary in sheep. After injection of the tracers in the retrochiasmatic area, which contains the cell group A15, fibres containing anterograde tracer were observed in the internal zone of the median eminence and in the pars nervosa of the pituitary. Numerous tyrosine hydroxylase immunoreactive fibres were present in the external zone of the median eminence and in the pars intermedia and the pars nervosa of the pituitary, with characteristic patterns of organisation in each area. Most tyrosine hydroxylase-immunoreactive fibres containing fluorescein dextran were located in the pars nervosa, whereas only a few were observed in the internal zone of the median eminence. It was concluded that at least part of the dopaminergic innervation of the pars nervosa originated from the A15 group. These results provide morphological evidence for (1) the role of dopaminergic neurons of the A15 cell group in the seasonal control of prolactin secretion via the release of dopamine in the pars nervosa, and (2) putative physiological interactions between dopamine and the secretion of neurohypophysial hormones in sheep.
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    Tyrosine hydroxylase in the cerebral ganglia of the American cockroach (Periplaneta americana L.): an immunohistochemical study (1995)
    Springer
    Cell & tissue research 282 (1995), S. 49-57 
    Details
    ISSN: 1432-0878
    Keywords: Tyrosine hydroxylase ; Catecholamine neurons ; Invertebrate nervous system ; Immunohistochemistry ; Cerebral ganglia ; Periplaneta americana (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have investigated the distribution of tyrosine-hydroxylase-like immunoreactivity in the cerebral ganglia of the American cockroach, Periplaneta americana. Groups of tyrosine-hydroxylase-immunoreactive cell bodies occur in various parts of the three regions of the cerebral ganglia. In the protocerebrum, single large neurons or small groups of neurons are located in the lateral neuropil, adjacent to the calyces, and in the dorsal portion of the pars intercerebralis. Small scattered cell bodies are found in the outer layers of the optic lobe, and clusters of larger cell bodies can be found in the deutocerebrum, medial and lateral to the antennal glomeruli. Thick bundles of tyrosine-hydroxylase-positive nerve fibers traverse the neuropil in the proto- and deutocerebrum and innervate the glomerular and the nonglomerular neuropil with fine varicose terminals. Dense terminal patterns are present in the medulla and lobula of the optic lobe, the pars intercerebralis, the medial tritocerebrum, and the area surrounding the antennal glomeruli, the central body and the mushroom bodies. The pattern of tyrosine-hydroxylase-like immunoreactivity is similar to that previously described for catecholaminergic neurons, but it is distinctly different from the distribution of histaminergic and serotonergic neurons.
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    Oxytocin-producing and vasopressin-producing eosinophils in the mouse spleen: immunohistochemical, immuno-electron-microscopic and in situ hybridization studies (1995)
    Springer
    Cell & tissue research 281 (1995), S. 1-10 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Spleen ; Oxytocin ; Vasopressin ; Immunohistochemistry ; Immuno-electron microscopy ; In situ hybridization ; Mouse (C57BL/6)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Oxytocin-like and vasopressin-like immunoreactive cells, and the cells expressing mRNAs for these peptides in the spleen of the C57BL/6 mouse were studied by immunohistochemistry, immuno-electron microscopy and in situ hybridization. Immunoreactive cells were distributed mainly in the splenic cord and marginal zone, whereas there were few in the lymphocyte-packed periarteriolar-lymphoid sheath, lymphoid follicle and germinal center. More numerous vasopressin-positive cells were seen in the splenic cord. The colocalization of oxytocin-like and vasopressin-like immunoreactivity in the same cells was identified by the investigation of mirror sections. By the pre-embedding immuno-electron-microscopic method using antisera against oxytocin and vasopressin, immunopositive reaction products were localized in the matrix around the specific granules, small clear vesicles and mitochondrial membrane of the eosinophils. No immunoreactivity to these peptides was found within the specific granules of the eosinophils. In situ hybridization with synthetic oligonucleotide probes labeled with 32P revealed the presence of mRNAs for oxytocin and vasopressin in the cells of the spleen, the distribution of the mRNAs for these peptides being the same as that of immunopositive cells. These observations suggest that eosinophils synthesize both oxytocin and vasopressin and store them in the matrix. Possible differences in the mechanism of synthesis and storage of these peptides between peripheral eosinophils and hypothalamic neurons are discussed.
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    Origins of nerve fibers containing nitric oxide synthase in the rat celiac-superior mesenteric ganglion (1995)
    Springer
    Cell & tissue research 281 (1995), S. 215-221 
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    ISSN: 1432-0878
    Keywords: Nitric oxide synthase ; Immunohistochemistry ; Retrograde tracing ; Celiac-superior mesenteric ganglion ; Sensory ganglion ; Spinal cord ; Intestine ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The origin of nitric oxide synthase-containing nerve fibers in rat celiac-superior mesenteric ganglion was examined using retrograde tracing techniques combined with the immunofluorescence method. Fluoro-Gold was injected into the celiac-superior mesenteric ganglion. Neuronal cell bodies retrogradely labeled with Fluoro-Gold in the thoracic spinal cord, the dorsal root ganglia at the thoracic level, the nodose ganglion, and the intestine from the duodenum to the proximal colon were examined for nitric oxide synthase immunoreactivity. About 60% of sympathetic preganglionic neurons in the intermediolateral nucleus projecting to the celiac-superior mesenteric ganglion were immunoreactive for nitric oxide synthase, as were approximately 27% of nodose ganglion neurons and about 65% of dorsal root ganglion neurons projecting to the cceliac-superior mesenteric ganglion. Neurons projecting to the celiac-superior mesenteric ganglion were found in the myenteric plexus of the small and large intestine. In the proximal colon, about 23% of such neurons were immunoreactive for nitric oxide synthase. However, in the small intestine, no immunoreactivity was found in these neurons.
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    Immunohistochemical localization of the calcium/calmodulin-dependent protein phosphatase, calcineurin, in the mouse testis: its unique accumulation in spermatid nuclei (1995)
    Springer
    Cell & tissue research 281 (1995), S. 273-281 
    Details
    ISSN: 1432-0878
    Keywords: Calcineurin ; Spermatogenesis ; Spermatids ; Nuclear transformation ; Immunohistochemistry ; Mouse (Jcl:ICR, BALB/c)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemical localization of a calmodulin-dependent protein phosphatase, calcineurin, was studied in the mouse testis in relation to previous observations showing that calmodulin is unusually rich in spermatogenic stages from mid-pachytene spermatocytes to elongating spermatids. The antibodies raised against calcineurin from scallop testis reacted with subunit B, but not subunit A, of calcineurin isoforms from mouse brain and testis. Indirect immunofluorescence using these antibodies on the mouse testis revealed positive reactions only in the nuclei of round or elongating spermatids: calcineurin started to accumulate in nuclei from the acrosomal cap phase, peaked at the initial stage of nuclear elongation, and decreased thereafter. There was almost no signal in the cytoplasm; spermatogenic cells at other stages, including spermatogonia, spermatocytes, mature sperm, and other somatic cells in the seminiferous tubules were totally negative. Immuno-electron microscopy gave the same result, on the basis of measuring the density of immunogold particles. These results suggest a role for calcineurin in remodeling of the nuclear chromatin in metamorphosing spermatids.
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    A temperature-sensitive lambda cl repressor functions on a modified operator in yeast cells by masking the TATA element (1995)
    Springer
    Molecular genetics and genomics 248 (1995), S. 499-505 
    Details
    ISSN: 1617-4623
    Keywords: α-Amylase ; Glyceraldehyde-3-phosphate-dehydrogenase promoter ; Phage lambda ; Repression ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We describe the construction and analysis of derivatives of the yeastTDH3 promoter in which the TATA box element has been replaced by a portion of the phage lambda operator containing a consensus TATA site flanked by binding sites for the cl repressor. Transcription of a reporter gene under the control of such a promoter is reduced in cells that express the cl repressor protein. Deletion of the native TATA element of theTDH3 promoter reduces transcription to the same extent. The cl repressor may act by “masking” the TATA element located between the repressor binding sites. Furthermore, the use of a temperature-sensitive cl repressor allowed temperature-dependent transcription of the reporter gene.
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    Isolation of temperature-sensitive mutants for mRNA capping enzyme in Saccharomyces cerevisiae (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 147-154 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; CEG1 ; Temperature-sensitive mutants ; mRNA capping enzyme ; Guanylyltransferase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The guanylyltransferase activity of mRNA capping enzyme catalyzes the transfer of GMP from GTP to the 5′ terminus of mRNA. In Saccharomyces cerevisiae, the activity is carried on the α subunit of capping enzyme, the product of the CEG1 gene. We have isolated 10 recessive, temperature-sensitive mutations of CEG1; nine (cegl-1 to cegl-9) were isolated on a single-copy plasmid and the remaining one (cegl-10) on a multicopy plasmid. The presence of cegl-10 in multiple copies is essential for the viability of cells carrying the mutation, and a shift to the restrictive temperature resulted in rapid growth arrest of cegl-10 cells, while growth rates of other mutants decreased gradually upon temperature upshift. Intragenic complementation was not observed for pairwise combinations of the mutations. Although the majority of the mutations occurred at the amino acid residues conserved between Cegl and the Schizosaccharomyces pombe homologue, none were located in the regions that are also conserved among viral capping enzymes and polynucleotide ligases. Guanylyltransferase activity of the mutant proteins as measured by covalent Ceg1-GMP complex formation was heat-labile. The availability of these mutants should facilitate studies of the structure-function relationships of capping enzyme, as well as the roles and regulation of mRNA capping.
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    URL: http://dx.doi.org/10.1007/BF00290360
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    Enhanced stability in vivo of a thermodynamically stable mutant form of yeast iso-1-cytochrome c (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 155-161 
    Details
    ISSN: 1617-4623
    Keywords: Cytochrome c ; Protein stability ; Protein degradation ; Mitochondria ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Previous work has established that the N57I amino acid replacement in iso-1-cytochrome c from the yeast Saccharomyces cerevisiae causes an unprecedented increase in thermodynamic stability of the protein in vitro, whereas the N57G replacement diminishes stability. Spectrophotometric measurements of intact cells revealed that the N57I iso-l-cytochrome c is present at higher than normal levels in vivo. Although iso-1-cytochrome c turnover is negligible during aerobic growth, transfer of fully derepressed, aerobically grown cells to anaerobic growth conditions leads to reduction in the levels of all of the cytochromes. Pulsechase experiments carried out under these anaerobic conditions demonstrated that the N57I iso-l-cytochrome c has a longer half-life than the normal protein. This is the first report of enhanced stability in vivo of a mutant form of a protein that has an enhanced thermodynamic stability in vitro. Although the N57I protein concentration is higher than the normal level, reduced growth in lactate medium indicated that the specific activity of this iso-l-cytochrome c in vivo is diminished relative to wild-type. On the other hand, the level of the thermodynamically labile N57G iso-1-cytochrome c was below normal. The in vivo levels of the N57I and N57G iso-l-cytochrome c suggest that proteins in the mitochondrial intermembrane space can be subjected to degradation, and that this degradation may play a role in controlling their normal levels.
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    A C-terminal region of the Saccharomyces cerevisiae transcription factor ADR1 plays an important role in the regulation of peroxisome proliferation by fatty acids (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 289-296 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Peroxisomes ; Catalase A ; ADR1 ; Peroxisome proliferation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Saccharomyces cerevisiae transcriptional activator ADR1, which controls ADH2 gene expression, was shown to be involved in the regulation of peroxisome proliferation. To study the mode of action of ADR1, we compared strains carrying the adr1-1 mutation, high or low copy numbers of the ADR1 gene, the constitutive allele ADR1-5 c, and 3′-deletions of ADR1. High ADR1 gene dosage increased the transcription of genes encoding peroxisomal proteins as compared to one copy of the ADR1 gene. Furthermore, overexpression of ADR1 under ethanol growth conditions induced the proliferation of peroxisomal structures. The organelles were observed to be localized in clusters, a typical feature of peroxisomes induced by oleic acid. In contrast, the ADR1-5 c allele, which induces ADH2 expression to a level comparable to that of high ADR1 gene dosage was found to have only a small effect. An analysis of functional domains of the ADR1 protein revealed that the N-terminal 220 amino acids of ADR1 were sufficient for wild-type levels of transcription of the FOX2, FOX3, and PAS1 genes, but the entire ADR1 protein was required for complete induction of the CTA1 gene and for growth oleic acid medium. Our data suggest that a functional domain of the ADR1 protein localized between residues 643 and 1323 is required for the induction of peroxisomal structures and for the utilization of oleic acid.
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    Cloning and characterization of seven cDNAs for hyperosmolarity-responsive (HOR) genes of Saccharomyces cerevisiae (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 127-138 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Hyperosmotic stress ; Signal transduction pathway
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Yeast cells can respond and adapt to osmotic stress. In our attempt to clarify the molecular mechanisms of cellular responses to osmotic stress, we cloned seven cDNAs for hyperosmolarity-responsive (HOR) genes from Saccharomyces cerevisiae by a differential screening method. Structural analysis of the clones revealed that those designated HOR1, HORS, HOR4, HOR5 and HOR6 encoded glycerol-3-phosphate dehydrogenase (Gpd1p), glucokinase (Glklp), hexose transporter (Hxtlp), heat-shock protein 12 (Hsp12p) and Na+, K+, Li+-ATPase (Enalp), respectively. HOR2 and HOR7 corresponded to novel genes. Gpdlp is a key enzyme in the synthesis of glycerol, which is a major osmoprotectant in S. cerevisiae. Cloning of HOR1/GPD1 as a HOR gene indicates that the accumulation of glycerol in yeast cells under hyperosmotic stress is, at least in part, caused by an increase in the level of GPDH protein. We performed a series of Northern blot analyses using HOR cDNAs as probes and RNAs prepared from cells grown under various conditions and from various mutant cells. The results suggested that all the HOR genes are regulated by common signal transduction pathways. However, the fact that they exhibited certain distinct responses indicated that they might also be regulated by specific pathways in addition to the common pathways. Ca2+ seemed to be involved in the signaling systems. In addition, Hog1p, one of the MAP kinases in yeast, appeared to be involved in the regulation of expression of HOR genes, although its function seemed to be insufficient for the overall regulation of expression of these genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00290358
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