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  • Immunocytochemistry  (90)
  • evolution  (81)
  • Springer  (171)
  • American Chemical Society
  • American Meteorological Society
  • 1995-1999
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  • 1993  (97)
  • 1991  (74)
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  • Springer  (171)
  • American Chemical Society
  • American Meteorological Society
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  • 1995-1999
  • 1990-1994  (171)
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  • 1
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    Springer
    Entomologia experimentalis et applicata 60 (1991), S. 173-182 
    ISSN: 1570-7458
    Keywords: Genetics ; evolution ; host adaptation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When populations are exposed to different environments, evolutionary processes can lead either to genetically differentiated strains or to the appearance of increased generalism at the individual level. For evolution to occur, genetic variability in performance in different environments is required. Here, intraspecific genetic variation across environments was estimated in the flour beetle Tribolium castaneum (Herbst) by comparing the responses of two strains of T. castaneum to different flour types. Replicated groups from each strain were allowed to develop on either the standard whole wheat medium or on one of four novel flours (wheat, rice, corn and oat). In several of the novel flours, clear differences in mean development time or population size of one or both strains were seen relative to performance in the standard medium. Moreover, the strains differed significantly in their phenotypic responses to the flours. One strain did particularly poorly on oat flour. Reduced oviposition, reduced larval survivorship and increased larval cannibalism were examined as possible causes of the low productivity on oat flour. These three factors accounted for about 70% of the reduction in population size when this strain oviposited and developed in oat flour. The difference between these two outbred strains in response to these five flours suggests that genetic variation in resource use is present within T. castaneum and may also be present within strains and natural populations in grain storage facilities. Such variation would permit an evolutionary response to selection in multiple environments (flours). This process has agricultural implications when several types of grain are stored in a single location because it could eventually lead to the evolution of highly generalized populations of T. castaneum, an important pest of stored products.
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  • 2
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    Journal of insect behavior 6 (1993), S. 715-735 
    ISSN: 1572-8889
    Keywords: Aphrodisiac ; cockroach ; evolution ; mating behavior ; sex pheromone ; sternal glands ; tergal glands
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two chemical signals are essential in all cockroach sexual behavioral sequences: the sex pheromone released by one partner, generally the female (for long distance attraction), and an aphrodisiac sex pheromone produced exclusively by male tergal glands (for female mounting and tergal contact or “feeding” behavior). Unlike the other cockroach groups, the males of the Oxyhaloinae species produce both chemical signals: the pheromone and the aphrodisiac. The occurrence of three patterns of mating behavior (A, B, and C), the production of male sex pheromones, and the existence in the male of developed sternal and tergal glands in seven related Oxyhaloinae species, make these cockroaches a useful model for studying the evolution of mating behavior patterns. The various types of mating behavior were not classified in the previous studies by Roth and Barth. In this report, they have been named type A (female in upper position), B (male in upper position), and C (male and female end to end). In type A mating, the male tergal glands, which are licked by the females, are well developed, whereas in types B and C, there is no licking of the male's tergal secretion by the females and the tergal glands are much less developed; the aphrodisiacs secreted by the tergal glands may no longer act in this case through contact chemoreception, but through an olfactory process involving volatile components. One common sex pheromone component seems to be acetoin. I suggest that the mating behavior tends from A toward B and C during the evolutionary process with a concomitant regression of the tergal glands and changes in the aphrodisiac emission levels. The mating behavioral sequences of cockroaches (Dictyoptera) and crickets (Orthoptera) show a striking degree of similarity and are probably examples of convergent evolution.
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  • 3
    ISSN: 1572-8889
    Keywords: Belostomatidae ; giant water bugs ; paternal care ; eggs ; reproduction ; behavior ; brooding ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Males of the giant water bug Lethocerus medius(Guerin) typify their monobasic subfamily, the Lethocerinae, in that they do not brood eggs attached to their backs as do males of all members of the subfamily Belostomatinae. Exclusive male parental investment as expressed in the Belostomatinae is extremely rare behavior among animals, and evolution of the trait is obscure. Lethocerus mediusmales apparently remain with their mates through oviposition and are consistently found in attendance of eggs after the female has departed. This behavior may enhance paternity assurance at no cost in opportunity for polygyny. Two double clutches of eggs were found, from which we infer the potential for polygynous matings and shared parental investment. Male L. mediusbrood attended egg clutches above the surface of the water, where they may moisten them, shade them, and defend them against predation. Egg attendance/brooding by L. mediusand other Lethocerusspecies may represent a plesiomorphic state from which paternal back- brooding evolved in the Belostomatinae.
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  • 4
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    Cellular and molecular life sciences 49 (1993), S. 1027-1036 
    ISSN: 1420-9071
    Keywords: Archaea (archaebacteria) ; extreme halophiles ; archaeol phospholipids ; archaeol glycolipids ; membrane function ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Archaebacteria (archaea) are comprised of three groups of prokaryotes: extreme halophiles, methanogens and thermoacidophiles (extreme thermophiles). Their membrane phospholipids and glycolipids are derived entirely from a saturated, isopranoid glycerol diether,sn-2,3-diphytanylglycerol (‘archaeol’) and/or its dimer, dibiphytanyldiglyceroltetraether (‘caldarchaeol’). In extreme halophiles, the major phospholipid is the archaeol analogue of phosphatidylglycerolmethylphosphate (PGP-Me); the glycolipids are sulfated and/or unsulfated glycosyl archaeols with diverse carbohydrate structure characteristic of taxons on the generic level. Biosynthesis of these archaeol-derived polar lipids occurs in a multienzyme, membrane-bound system that is absolutely dependent on high salt concentration (4 M). The highly complex biosynthetic pathways involve intermediates containing glycerol ether-linked C20-isoprenyl groups which are reduced to phytanyl groups to give the final saturated polar lipids. In methanogens, polar lipids are derived both from archaeol and caldarchaeol, and thermoacidophiles contain essentially only caldarchaeol-derived polar lipids. The function of these membrane polar lipids in maintaining the stability, fluidity and ionic properties of the cell membrane of extreme halophiles, as well as the evolutionary implications of the archaeol and caldarchaeol-derived structures will be discussed.
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  • 5
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    Cellular and molecular life sciences 49 (1993), S. 317-319 
    ISSN: 1420-9071
    Keywords: Chitin ; cuticle ; evolution ; vertebrates ; bony fish ; Blenniidae ; Paralipophrys trigoides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Lectin binding, endo-chitinase binding and enzymatic degradation studies show that the epidermal cuticle of the bony fishParalipophrys trigloides (Blenniidae) is chitinous. This is the first evidence that a vertebrate species possesses a chitinous tissue. Recently aXenopus gene has been identified which has significant sequence similarity to the catalytic domain of yeast chitin synthase III, a chitin producing enzyme1,2. Taken together these two findings imply that chitin synthesis capability may be a basic vertebrate feature.
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  • 6
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    Journal for general philosophy of science 22 (1991), S. 133-141 
    ISSN: 1572-8587
    Keywords: evolution ; teleology ; chance ; purpose ; anthropomorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Philosophy , Nature of Science, Research, Systems of Higher Education, Museum Science
    Notes: Summary Revaluation of the problem of natural teleology seems an important precondition for elucidating our environmental crisis and for formulating an ‘ecological ethics’, because it calls for a recognition of an intrinsic value in nature and organisms. Therefore, it is necessary to show that the concept of natural teleology is not in contradiction with scientific theories, in particular not with the theory of evolution. In this paper I shall argue that there is a fundamental misunderstanding about the concepts of teleology and chance in modern thinking. This as a result of a radical transformation of the Aristotelian concept of teleology by Christian theologians during the Middle Ages. This confusion resulted in the rejection of teleology from evolution and in an exaggeration of the role of chance. However, not a solution for the problem of teleology is given here, but only an attempt to prove that neither the fossil-record, nor the role of chance in evolution can give adequate arguments for the negation of teleology in evolution. That is not to say that, therefore there exists teleology in evolution, but the problem of teleology in nature cannot, be solved by the scientific theory of evolution, but only be elucidated by philosophical analysis. At the end of the paper it is argued that teleology must be rather presupposed in evolution.
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  • 7
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    Entomologia experimentalis et applicata 66 (1993), S. 3-12 
    ISSN: 1570-7458
    Keywords: evolution ; coevolution ; selection ; insect attack ; plant defense ; competition ; enemy free space ; chemoreception ; specialization ; plant recognition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Most hypotheses concerning the evolution of insect-plant relationships are based on the assumptions that, (1) phytophagous insects reduce plant fitness, and that (2) insect-plant relationships are the result of unconstrained selection. It can be shown, however, that there is little evidence to support these assumptions. As an alternative, it is proposed that the evolution of insect-plant relationships results primarily from autonomous evolutionary events; namely from heritable functional changes within the insects' nervous system that determine plant recognition and ultimately host plant specificity. These changes cannot be evoked by selective ecological agents. They originate from intrinsic changes (mutationssensu lato) within the insect genome. Ecological factors play a secondary role: by either supporting or preventing the establishment of the new genotype with the novel food preference.
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  • 8
    ISSN: 1432-1351
    Keywords: Photoreception ; Extraretinal Photoreceptor ; Chromophore ; Opsin ; Reptile ; Immunocytochemistry ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Since the beginning of this century evidence has accumulated which demonstrates that non-mammalian vertebrates possess photoreceptors situated deep within the brain. While many attempts have been made to localize these sensory cells, studies have either failed or been inconclusive. In this report we have used several experimental approaches to localize the deep brain photoreceptors of the lizard Anolis carolinensis. Using 3 antibodies that bind vertebrate cone opsins, we have immunolabelled cerebrospinal fluid (CSF)-contacting neurons located at the ventricular border within the nucleus ventromedialis of the septum. Western blot analysis indicates that these antibodies recognized a single 40 kD protein in ocular, anterior brain, and pineal extracts. Immunoblots of rodent brain did not show a similar protein band. We have also identified specific retinoids associated with phototransduction (11-cis and all-trans-3,4-didehydroretinaldehyde) within anterior brain extracts. This combined data provides the most detailed analysis of deep brain photoreceptors in any vertebrate. Consequently, we feel Anolis provides an excellent model to study this unexplored sensory system of the vertebrates.
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  • 9
    ISSN: 1432-1351
    Keywords: Egg laying hormone ; Aplysia ; Binding protein ; Immunocytochemistry ; Reproductive system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A protein solubilized from a membrane preparation of the gonad of Aplysia californica has been isolated by affinity chromatography, using bag cell egg-laying hormone (ELH) as the bound ligand, and partially purified and characterized by gel electrophoresis. The protein has an apparent molecular weight of 52 kDa and consists of two disulfide-linked subunits of about 30 kDa each. The protein is glycosylated and has an acidic pI. Approximately 10–15 μg of this protein can be isolated from a single ovotestis, representing less than 1% of the total protein in the gonad; but the protein could not be detected in buccal mass or body wall, tissues which do not have apparent response to ELH. Antibodies generated against this ELH-binding protein (ELHBP) were used to localize sites in the ovotestis which might contain this molecule and thus represent targets for egg-laying hormone. Immunocytochemical results indicate that the oocytes are a rich source of this protein, since their cytoplasm was the only detectable site of immunoreactivity. Whether this binding protein represents an egg-laying hormone receptor is uncertain, but its prevalence in oocytes suggests that ELH plays a signaling role on these gametes.
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  • 10
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    Insectes sociaux 38 (1991), S. 263-272 
    ISSN: 1420-9098
    Keywords: Isoptera ; Kalotermitidae ; Neotermes papua ; termites ; caste differentiation ; division of labour ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The direct development ofNeotermes papua (Isoptera: Kalotermitidae) comprises four larval and three nymphal instars before the alate. The first five instars can be easily characterized. The second stage nymphs come morphologically close to the pseudergates, characterized by reduced wing buds. These nymphs can moult stationarily, i.e. with little morphological change, or to presoldiers, or proceed to the alate via the third nymphal stage. Pseudergates originate through a late and reversible deviation from the straight development to the alate. Presoldiers may derive from several stages, up to the last nymphal one; their production is subject to an inhibition by extant soldiers. This developmental schema is congruent with those described in other Kalotermitidae and the Termopsidae. By pinpointing the existence of a large pool of pluripotent individuals, in which the penultimate nymphal stage mingles with pseudergates, the present study also reveals a great similarity betweenNeotermes andProrhinotermes, and suggests that this developmental schema might be generally applicable to termites devoid of a permanent worker caste.
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  • 11
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    Insectes sociaux 40 (1993), S. 325-335 
    ISSN: 1420-9098
    Keywords: Formicidae ; social parasitism ; PCR ; 18 S ribosomal RNA ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The evolutionary relationship between socially parasitic ants and their hosts is still an unsolved problem. We have compared a 1.2 kb sequence of the 18 S ribosomal RNA genes of the parasitic antsDoronomyrmex kutteri, Harpagoxenus sublaevis andChalepoxenus muellerianus to the sequence of the host speciesLeptothorax acervorum andL. recedens (all subfamily Myrmicinae, tribe Leptothoracini) and to an out-group antCamponotus ligniperda (Formicinae). We found that parasitic species and the host species and alsoCamponotus ligniperda differ at less than 1% of the base positions of the 1.2 kb segment of the 18S rRNA gene. The sequences showed 80.3% identity to the 18 S ribosomal RNA genes of the beetleTenebrio molitor and only 66.5% to that of the dipteranDrosophila melanogaster.
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  • 12
    ISSN: 1423-0445
    Keywords: defensive secretion ; hot secretion ; elytral flanges ; evolution ; benzoquinones ; hydrocarbons ; bombardier beetle ; Coleoptera ; Carabidae ; Paussinae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Australian bombardier beetle,Mystropomus regularis, sprays a mixture of quinones (1,4-benzoquinone, 2-methyl-1,4-benzoquinone, 2-ethyl-1,4-benzoquinone) and hydrocarbons (principallyn-pentadecane). The defensive fluid ist generated explosively in two-chambered glands, and is ejected audibly and hot (maximal recorded temperature = 59°C).Mystropomus is a member of the paussoid lineage of bombardiers. In common with other members of the group, it has a pair of elytral flanges (flanges of Coanda), associated with the gland openings, that serve as launching guides for anteriorly-aimed ejections of spray. It is argued thatMystropomus may be the least derived of flanged paussoids, and the closest living relative of the most primitive of extant bombardiers (Metriini).
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  • 13
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    Cellular and molecular life sciences 47 (1991), S. 791-803 
    ISSN: 1420-9071
    Keywords: Phytotoxins ; ecology ; phylogeny ; evolution ; biosynthesis ; coronatine ; phaseolotoxin ; rhizobitoxine ; syringomycin ; syringotoxin ; syringostatin ; tabtoxin ; tagetitoxin ; tropolone ; fireblight toxin ; thaxtomin ; 3-methylthiopropionic acid ; carboxylic acids ; Pseudomonas ; Xanthomonas ; Xanthomonas ; Streptomyces ; Erwinia ; Bradyrhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract This review attempts to rationalise what is known about bacterial phytotoxins and associate it with the ecology and possible evolution of the producing organisms. Study of non-toxin producing variants gives insight into the ecological role of the toxin. Elucidation of chemical structures of phytotoxins has shown that many exist as families of analogous compounds. Studies on the variation of chemical structures and how they are distributed across species and genera can lead to development of hypotheses on evolutionary relationships. Knowledge on biosynthetic pathways to tosins allows recognition of specific enzymatic steps involved in developing the characteristic features of the structures. Phytotoxins often have a potent biochemical activity, and in some cases the producing organism has associated mechanisms to prevent action of the toxin upon itself; in such cases toxigenesis is clearly not a chance event. The various aspects of bacterial toxigenesis indicate that bacterial phytotoxins are special secondary metabolic products that play beneficial roles to the producing organisms in their various ecological niches.
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  • 14
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    Journal of comparative physiology 169 (1991), S. 39-50 
    ISSN: 1432-1351
    Keywords: Photoreception ; Retinally degenerate ; Mouse ; Circadian ; Rods ; Cones ; 11-cis retinaldehyde ; Immunocytochemistry ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We have examined the effects of light on circadian locomotor rhythms in retinally degenerate mice (C57BL/6J mice homozygous for the rd allele: rd/rd). The sensitivity of circadian photoreception in these mice was determined by varying the irradiance of a 15 min light pulse (515 nm) given at circadian time 16 and meauring the magnitude of the phase shift of the locomotor rhythm. Experiments were performed on animals 80 days of age. Despite the loss of visual photoreceptors in the rd/rd retina, animals showed circadian responses to light that were indistinguishable from mice with normal retinas (rd/+ and +/+). While no photoreceptor outersegments were identified in the retina of rd/rd animals (80–100 days of age), we did identify a small number of perikarya that were immunoreactive for cone opsins, and even fewer cells that contained rod opsin. Using HPLC, we demonstrated the presence and photoisomerization of the rhodopsin chromophore 11-cis retinaldehyde. The rd/rd retinas contained about 2% of 11-cis retinaldehyde found in +/+ retinas. We have yet to determine whether the opsin immunoreactive perikarya or some other unidentified cell type mediate circadian light detection in the rd/rd retina.
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  • 15
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    Plant molecular biology 23 (1993), S. 409-413 
    ISSN: 1573-5028
    Keywords: cDNA ; cloning ; rice ; L5 ; ribosomal 5 S RNA-binding protein ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A rice (Oryza sativa L.) cDNA clone coding for the cytoplasmic ribosomal protein L5, which associates with 5 S rRNA for ribosome assembly, was cloned and its nucleotide sequence was determined. The primary structure of rice L5, deduced from the nucleotide sequence, contains 294 amino acids and has intriguing features some of which are also conserved in other eucaryotic homologues. These include: four clusters of basic amino acids, one of which may serve as a nucleolar localization signal; three repeated amino acid sequences; the conservation of glycine residues. This protein was identified as the nuclear-encoded cytoplasmic ribosomal protein L5 of rice by sequence similarity to other eucaryotic ribosomal 5 S RNA-binding proteins of rat, chicken, Xenopus laevis, and Saccharomyces cerevisiae. Rice L5 shares 51 to 62% amino acid sequence identity with the homologues. A group of ribosomal proteins from archaebacteria including Methanococcus vanniellii L18 and Halobacterium cutirubrum L13, which are known to be associated with 5 S rRNA, also related to rice L5 and the other eucaryotic counterparts, suggesting an evolutionary relationship in these ribosomal 5 S RNA-binding proteins.
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  • 16
    ISSN: 1573-5028
    Keywords: cDNA sequences ; evolution ; fructose-1,6-bisphosphate aldolase ; Spinacia oleracea ; transit peptide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report the sequences of full-length cDNAs for the nuclear genes encoding the chloroplastic and cytosolic fructose-1,6-bisphosphate aldolase (EC 4.1.2.13) from spinach. A comparison of the deduced amino-acid sequences with one another and with published cytosolic aldolase sequences of other plants revealed that the two enzymes from spinach share only 54% homology on their amino acid level whereas the homology of the cytosolic enzyme of spinach with the known sequences of cytosolic aldolases of maize, rice and Arabidopsis range from 67 to 92%. The sequence of the chloroplastic enzyme includes a stroma-targeting N-terminal transit peptide of 46 amino acid residues for import into the chloroplast. The transit peptide exhibits essential features similar to other chloroplast transit peptides. Southern blot analysis implies that both spinach enzymes are encoded by single genes.
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  • 17
    ISSN: 1573-5028
    Keywords: C4 metabolism ; evolution ; GC content ; gene family ; PEPC ; Sorghum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Although housekeeping functions have been shown for the phosphoenolpyruvate carboxylase (EC 4.1.1.31, PEPC) in plants and in prokaryotes, PEPC is mainly known for its specific role in the primary photosynthetic CO2 fixation in C4 and CAM plants. We have shown that in Sorghum, a monocotyledonous C4 plant, the enzyme is encoded in the nucleus by a small multigene family. Here we report the entire nucleotide sequence (7.5 kb) of the third member (CP21) that completes the structure of the Sorghum PEPC gene family. Nucleotide composition, CpG islands and GC content of the three Sorghum PEPC genes are analysed with respect to their possible implications in the regulation of expression. A study of structure/function and phylogenetic relationships based on the compilation of all PEPC sequences known so far is presented. Data demonstrate that (1) the different forms of plant PEPC have very similar primary structures, functional and regulatory properties, (2) neither apparent amino acid sequences nor phylogenetic relationships are specific for the C4 and CAM PEPCs and (3) expression of the different genes coding for the Sorghum PEPC isoenzymes is differently regulated (i.e. by light, nitrogen source) in a spatial and temporal manner. These results suggest that the main distinguishing feature between plant PEPCs is to be found at the level of genes expression rather than in their primary structure.
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  • 18
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    Biochemical genetics 31 (1993), S. 473-484 
    ISSN: 1573-4927
    Keywords: hydroxyacyl glutathione hydrolase (glyoxalase II) ; chromosome mapping ; evolution ; Mus musculus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract In man, the gene for hydroxyacyl glutathione hydrolase (HAGH; glyoxalase II) is closely linked to the α-globin locus (HBα) on Chromosome 16. HAGH polymorphism in the mouse has now enabled the mapping of the murine homologue. Deletion mapping, congenic strain studies, and characterization of 41 recombinant inbred strains establish that the mouseHagh locus lies very close to the α-globin pseudogene (Hba-ps4) in the vicinity of the major histocompatibility locus (H-2) on chromosome 17. Several other loci have been identified previously that are also closely linked to the human α-globin locus but near the α-globin pseudogeneHba-ps4 in the mouse. These linkage relationships suggest that during the evolution of mice a translocation occurred that subdivided the α-globin locus, leaving one inactive α-globin gene still associated with theHagh locus and linked sequences, while moving and inserting the active α-globin locus and all distal sequences into an internal location on another autosome, the predecessor to mouse chromosome 11.
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  • 19
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    Biochemical genetics 31 (1993), S. 329-341 
    ISSN: 1573-4927
    Keywords: actin superfamily ; Drosophila genetics ; ATPase domain ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Diverse proteins that are 35% to 55% identical to actins have been discovered recently in yeasts, nematodes, and vertebrates. In order to study these proteins systematically and relate their functions to those of conventional actins, we are isolating the corresponding genes from the genetically tractable eukaryote,Drosophila melanogaster. Here we report the isolation and partial characterization of aDrosophila homologue of theSchizosaccharomyces pombe act2 gene. Degenerate oligonucleotide primers specifying peptides that are highly conserved within the actin protein superfamily were used in conjunction with polymerase chain reaction (PCR) to amplify a portion of theDrosophila gene that we have namedactr66B. The corresponding full-length cDNA sequence encodes a protein of 418 residues that is 65% identical to the product of theS. pombe act2 gene, 80% identical to the bovineact2 homologue, but only 48% identical to the principalDrosophila cytoplasmic actin encoded by theAct5C actin gene. Alignment of the yeast, bovine, andDrosophila actin-related proteins shows that they have four peptide insertions, relative to conventional actins, three of which are well placed to modify actin polymerization and one that is likely to perturb the binding of myosin. Locations of two of the fiveactr66B introns are conserved betweenDrosophila and yeast genes, further attesting that they evolved from a common ancestor and are likely to encode proteins having similar functions. We demonstrate that theDrosophila gene is located on the left arm of chromosome 3, within subdivision 66B. Finally, we show by RNA blot-hybridization that the gene is expressed at low levels, relative to conventional nonmuscle actin, in all developmental stages. From these and other observations we infer that the actr66B protein is a minor component of all cells, perhaps serving to modify the polymerization, structure, and dynamic behavior of actin filaments.
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  • 20
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    Biochemical genetics 31 (1993), S. 473-484 
    ISSN: 1573-4927
    Keywords: hydroxyacyl glutathione hydrolase (glyoxalase II) ; chromosome mapping ; evolution ; Mus musculus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract In man, the gene for hydroxyacyl glutathione hydrolase (HAGH; glyoxalase II) is closely linked to the α-globin locus (HBα) on Chromosome 16. HAGH polymorphism in the mouse has now enabled the mapping of the murine homologue. Deletion mapping, congenic strain studies, and characterization of 41 recombinant inbred strains establish that the mouseHagh locus lies very close to the α-globin pseudogene (Hba-ps4) in the vicinity of the major histocompatibility locus (H-2) on chromosome 17. Several other loci have been identified previously that are also closely linked to the human α-globin locus but near the α-globin pseudogeneHba-ps4 in the mouse. These linkage relationships suggest that during the evolution of mice a translocation occurred that subdivided the α-globin locus, leaving one inactive α-globin gene still associated with theHagh locus and linked sequences, while moving and inserting the active α-globin locus and all distal sequences into an internal location on another autosome, the predecessor to mouse chromosome 11.
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  • 21
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    Biochemical genetics 31 (1993), S. 329-341 
    ISSN: 1573-4927
    Keywords: actin superfamily ; Drosophila genetics ; ATPase domain ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Diverse proteins that are 35% to 55% identical to actins have been discovered recently in yeasts, nematodes, and vertebrates. In order to study these proteins systematically and relate their functions to those of conventional actins, we are isolating the corresponding genes from the genetically tractable eukaryote,Drosophila melanogaster. Here we report the isolation and partial characterization of aDrosophila homologue of theSchizosaccharomyces pombe act2 gene. Degenerate oligonucleotide primers specifying peptides that are highly conserved within the actin protein superfamily were used in conjunction with polymerase chain reaction (PCR) to amplify a portion of theDrosophila gene that we have namedactr66B. The corresponding full-length cDNA sequence encodes a protein of 418 residues that is 65% identical to the product of theS. pombe act2 gene, 80% identical to the bovineact2 homologue, but only 48% identical to the principalDrosophila cytoplasmic actin encoded by theAct5C actin gene. Alignment of the yeast, bovine, andDrosophila actin-related proteins shows that they have four peptide insertions, relative to conventional actins, three of which are well placed to modify actin polymerization and one that is likely to perturb the binding of myosin. Locations of two of the fiveactr66B introns are conserved betweenDrosophila and yeast genes, further attesting that they evolved from a common ancestor and are likely to encode proteins having similar functions. We demonstrate that theDrosophila gene is located on the left arm of chromosome 3, within subdivision 66B. Finally, we show by RNA blot-hybridization that the gene is expressed at low levels, relative to conventional nonmuscle actin, in all developmental stages. From these and other observations we infer that the actr66B protein is a minor component of all cells, perhaps serving to modify the polymerization, structure, and dynamic behavior of actin filaments.
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  • 22
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; Ti plasmid ; evolution ; IS elements ; host range
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The TA regions of biotype III octopine/cucumopine (OC) Ti plasmids are closely related to the TL region of the biotype I octopine Ti plasmids pTiAch5 and pTi15955. Sequence analysis shows that the limited and wide host range biotype III OC TA regions are derived from a common ancestor structure which lacked the 6a gene found in the biotype I octopine TL region. The TA region of the wide host range OC Ti plasmids has conserved most of the original TL-like structure. In most wide host range OC isolates the TA-iaaH gene is inactivated by the insertion of an IS866 element. However, the TA region of the wide host range isolate Hm1 carries an intact TA-iaaH gene. This gene encodes a biologically active product, as shown by root induction tests and indole-3-acetic acid measurements. The limited host range OC Ti plasmids pTiAB3 and pTiAg57 have shorter TA regions which are derived from a wide host range TA region. The AB3 type arose by an IS868-mediated, internal TA region deletion which removed the iaa genes and part of the ipt gene and left a copy of IS868 at the position of the deleted fragment. The pTiAB3 iaa/ipt deletion was followed by insertion of a second IS element, IS869, immediately 3′ of the ipt gene. pTiAg57 underwent the same iaa-ipt deletion as pTiAB3, but lacks the IS868 and IS869 elements. Analysis of the various TA region structures provides a detailed insight into the evolution of the biotype III OC strains.
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  • 23
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    Plant molecular biology 16 (1991), S. 335-337 
    ISSN: 1573-5028
    Keywords: Triticum ; wheat ; endosperm ; gliadin ; pseudogene ; duplication ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 24
    ISSN: 1573-5028
    Keywords: calcium-binding protein ; centrin ; EF hand ; evolution ; green algae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Centrin (= caltractin) is a ubiquitous, cytoskeletal protein which is a member of the EF-hand superfamily of calcium-binding proteins. A centrin-coding cDNA was isolated and characterized from the prasinophyte green alga Scherffelia dubia. Centrin PCR amplification primers were used to isolate partial, homologous cDNA sequences from the green algae Tetraselmis striata and Spermatozopsis similis. Annealing analyses suggested that centrin is a single-copy-coding region in T. striata and S. similis and other green algae studied. Centrin-coding regions from S. dubia, S. similis and T. striata encode four colinear EF-hand domains which putatively bind calcium. Phylogenetic analyses, including homologous sequences from Chlamydomonas reinhardtii and the land plant Atriplex nummularia, demonstrate that the domains of centrins are congruent and arose from the two-fold duplication of an ancestral EF hand with Domains 1+3 and Domains 2+4 clustering. The domains of centrins are also congruent with those of calmodulins demonstrating that, like calmodulin, centrin is an ancient protein which arose within the ancestor of all eukaryotes via gene duplication. Phylogenetic relationships inferred from centrin-coding region comparisons mirror results of small subunit ribosomal RNA sequence analyses suggesting that centrin-coding regions are useful evolutionary markers within the green algae.
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  • 25
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    Plant molecular biology 16 (1991), S. 537-546 
    ISSN: 1573-5028
    Keywords: Antithamnion sp. ; chloroplast ; Cyanidium caldarium Geitler ; evolution ; red algae ; rRNA operon ; rRNA spacer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nucleotide sequences of the plastid 16S rDNA of the multicellular red alga Antithamnion sp. and the 16S rDNA/23S rDNA intergenic spacers of the plastid DNAs of the unicellular red alga Cyanidium caldarium and of Antithamnion sp. were determined. Sequence comparisons support the idea of a polyphyletic origin of the red algal and the higher-plant chloroplasts. Both spacer regions include the unsplit tRNAIle (GAU) and tRNAAla (UGC) genes and so the plastids of both algae form a homogeneous group with those of chromophytic algae and Cyanophora paradoxa characterized by ‘small-sized’ rDNA spacers in contrast to green algae and higher plants. Nevertheless, remarkable sequence differences within the rRNA and the tRNA genes give the plastids of Cyanidium caldarium a rather isolated position.
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  • 26
    ISSN: 1573-5028
    Keywords: algae ; endosymbiosis ; evolution ; plastidial DNA ; Rubisco operon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The rbcS gene coding for the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) of the brown alga Pylaiella littoralis is located within the plastid genome and is transcribed as a single polycistronic mRNA with the gene for the large subunit of Rubisco, rbcL. The structure of the Rubisco operon from P. littoralis was determined. Molecular phylogenies for rbcS and rbcL with a wide range of prokaryotes and eukaryotes were constructed which are congruent with recent evidence for polyphyletic plastid origins. Both rbcL and rbcS of the β-purple bacterium Alcaligenes eutrophus clearly cluster with the rhodophyte and chromophyte proteins. The data suggest that the Rubisco operons of red algal and chromophytic plastids derive from β-purple eubacterial antecedents, rather than the cyanobacterial lineage of eubacteria from which other of their genes derive. This implies a lateral transfer of Rubisco genes from β-purple eubacterial ancestors to the cyanobacterial ancestor of rhodophyte and chromophyte plastids.
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  • 27
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    Plant molecular biology 17 (1991), S. 641-652 
    ISSN: 1573-5028
    Keywords: F-ATPase ; cyanobacteria ; operon ; evolution ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The two operons atp1 and atp2, encoding the subunits of the FOF1 ATP-synthase, have been cloned and sequenced from the cyanobacterium Synechocystis sp. PCC 6803. The organization of the different genes in the operons have been found to resemble that of the cyanobacteria Synechococcus sp. PCC 6301 and Anabaena sp. PCC 7120. The Synechocystis FOF1 ATP-synthase has nine subunits. A tenth open reading frame with unknown function was detected at the 5′ end of atp1, coding for a putative gene product similar to uncI in Escherichia coli. A promoter structure was inferred for the Synechocystis atp operons and compared to other known promoters of cyanobacteria. Even though the operon structure of atp1 and atp2 in Synechocystis resembles the corresponding operons of Synechococcus, the amino acid sequences of individual gene products show marked differences. Genetic distances between cyanobacterial genes and genes for ATP-synthase subunits from other species have been calculated and compiled into evolutionary trees.
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  • 28
    ISSN: 1573-5028
    Keywords: Rubisco activase ; rca ; rbcLrbcS ; cyanobacteria ; expression ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A gene encoding ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase (rca) was found downstream from the rbcLrbcS operon in the heterocystous cyanobacterium Anabaena sp. strain CA. Two unknown open reading frames were shown to be located between rbcS and rca in strain CA and all the genes, rbcLrbcS, ORF1, ORF2, and rca were in the same transcriptional orientation. The deduced amino acid sequence of the Anabaena Rubisco activase showed both similarities and differences to the plant enzyme with considerable differences at the carboxy and amino termini. Proposed ATP-binding sites were conserved in the cyanobacterial protein. Recombinant cyanobacterial Rubisco activase, however, reacted with antisera to spinach Rubisco activase. Hybridization studies, using the Anabaena sp. strain CA rca gene as a heterologous probe, detected homologous sequences in heterocystous Anabaena/Nostoc strains but not in unicellular or nonheterocystous filamentous cyanobacteria, suggestive of a close evolutionary relationship of chloroplasts and heterocystous cyanobacteria.
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  • 29
    ISSN: 1432-1351
    Keywords: Aplysia ; Motoneurons ; Immunocytochemistry ; Small cardioactive peptides ; Facilitation ; Depression ; Buccal ; Feeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We used physiological recordings, intracellular dye injections and immunocytochemistry to further identify and characterize neurons in the buccal ganglia of Aplysia calif ornica expressing Small Cardioactive Peptide-like immunoreactivity (SCP-LI). Neurons were identified based upon soma size and position, input from premotor cells B4 and B5, axonal projections, muscle innervation patterns, and neuromuscular synaptic properties. SCP-LI was observed in several large ventral neurons including B6, B7, B9, B10, and B11, groups of s1 and s2 cluster cells, at least one cell located at a branch point of buccal nerve n2, and the previously characterized neurons B1, B2 and B15. B6, B7, B9, B10 and B11 are motoneurons to intrinsic muscles of the buccal mass, each displaying a unique innervation pattern and neuromuscular plasticity. Combined, these motoneurons innervate all major intrinsic buccal muscles (I1/I3, I2, I4, I5, I6). Correspondingly, SCP-LI processes were observed on all of these muscles. Innervation of multiple nonhomologous buccal muscles by individual motoneurons having extremely plastic neuromuscular synapses, represents a unique form of neuromuscular organization which is prevalent in this system. Our results show numerous SCPergic buccal motoneurons with widespread ganglionic processes and buccal muscle innervation, and support extensive use of SCPs in the control of feeding musculature.
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  • 30
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    Cell & tissue research 271 (1993), S. 107-113 
    ISSN: 1432-0878
    Keywords: Proventriculus ; Endocrine ontogenesis ; Ultrastructure ; Regulatory peptides ; Immunocytochemistry ; Silver impregnations ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The development of endocrine cells in the chicken proventriculus has been investigated using light-and electron-microscopy in conjunction with silver and immunocytochemical techniques. The first morphologically detectable endocrine cells were found in 5-day-old embryos by electron microscopy. From the 9th to the 13th day, endocrine cells in contact with the lumen of the organ could be detected both by electron and light (silver impregnation) microscopy. The number of open-type endocrine cells progressively decreased and the number of closed-type increased after this stage. Until the 16th day, endocrine cells were located exclusively in the luminal epithelium, but afterwards they appeared in progressively greater numbers in the compound glands. After hatching, long cytoplasmic processes could be seen in the endocrine cells. Immunoreactivities to regulatory substances appeared in the following order: serotonin (day-14), avian pancreatic polypeptide, glucagon and somatostatin (day-16), bombesin and neurotensin (day-18), and finally, met-enkephalin (day-21).
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  • 31
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    Cell & tissue research 272 (1993), S. 161-168 
    ISSN: 1432-0878
    Keywords: Keratinocyte subpopulations ; Epidermal calcium-binding protein ; Low gravity sedimentation ; Immunoblotting ; Immunocytochemistry ; Flow cytometry ; Rats (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Epidermal calcium-binding protein (ECaBP) is present in the cells of the basal layer of the epidermis and other stratified epithelia. Since the basal layer compartment contains at least two types of cells: slow-cycling, poorly-differentiated, and actively proliferating, more differentiated cells, it was of interest to determine whether they both contained ECaBP. Basal and nearly suprabasal layer keratinocytes from newborn rat epidermis were fractionated into three fractions on the basis of cell size, using low-gravity sedimentation. The cell differentiation in each subgroup was estimated by cell size, morphology, cell cycle stage, RNA/DNA content, and the presence of specific keratins. The presence of ECaBP in these fractions was detected by immunocytochemistry and immunoblotting. Double staining with ECaBP antibodies and propidium iodide followed by flow cytometry was used to correlate ECaBP production and the stage of cell cycle. The relative cell size, measured by the light scattering was used to study the relationship between cell size and ECaBP production. The results show that small keratinocytes with low DNA and RNA content (G0 cells) do not express ECaBP. ECaBP was found only in intermediate size basal keratinocytes with higher DNA and RNA contents, corresponding to actively proliferating S phase cells. Large keratinocytes, which express suprabasal keratin and have low DNA and high RNA content, cease to express ECaBP. ECaBP may, therefore, be a useful marker for assessing the movement of cells from poorly differentiated reserve compartment towards proliferation and further differentiation in both physiological and pathological situations.
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  • 32
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    Cell & tissue research 263 (1991), S. 207-215 
    ISSN: 1432-0878
    Keywords: Somatolactin (SL) ; Pituitary gland, pars intermedia ; PAS-positive cells ; Immunocytochemistry ; Gadus morhua, Oncorhynchus mykiss, Poecillia latipinna (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We report here on the cellular localization in the fish pituitary of somatolactin (SL), a putative new pituitary hormone related to growth hormone and prolactin, which has been recently identified in the piscine pituitary gland. Immunocytochemical staining, using anti-cod SL serum, revealed that in the cod pituitary gland, SL is produced by cells in the intermediate lobe, bordering the neural tissue. These cells, staining weakly with periodic-acid-Schiff (PAS), are distinct from the melanocyte stimulating hormone (MSH) cells which, as in all teleosts, are PAS-negative. SL-immunoreactivity was observed in the same location in all other teleost species examined: flounder, rainbow trout, killifish, molly, catfish and eel. In most fish the SL-immunoreactive cells are either strongly or weakly PAS-positive but in rainbow trout are chromophobic, indicating that the SL protein can probably exist in glycosylated and non-glycosylated forms. Thus, in demonstrating the cellular localization of SL, this study provides the first identification of the enigmatic, second cell-type of the fish pars intermedia.
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  • 33
    ISSN: 1432-0878
    Keywords: Spinal cord ; Motoneurones ; Dorsal horn ; Neurofilament ; Phosphorylation ; Immunocytochemistry ; Rat ; Cat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Neurofilament immunoreactivity was examined in spinal cords of rats and cats with antibodies to all three subunits (68 kD, 155 kD and 200 kD) and to different phosphorylation states of 200 kD. NFHP-, an antibody against non-phosphorylated 200 kD, labelled all rat neuronal perikarya but failed to labet cat neurofilaments. In both species, the perikarya and processes of motoneurones were immunoreactive for all three subunits but most dorsal horn neuronal perikarya were not immunoreactive for 68 kD and 155 kD. Motoneuronal perikarya and proximal processes showed filamentous labelling for 68 kD but not for 155 kD in the rat, while in neither species did these show labelling with RT97, an antibody against a highly phosphorylated form of 200 kD; immunoreactivity for 200 kD was present in both filamentous (probably partially phosphorylated) and non-filamentous (non-phosphorylated) forms, but in dorsal horn neurones only the latter was present. Interpretations consistent with this data are: in rat and possibly also cat, motoneuronal neurofilaments consist of a 68 kD backbone with partially phosphorylated 200 kD sidearms, with both 155 kD and 200 kD (non-phosphorylated) subunits in a non-filamentous form; this neurofilament becomes more highly phosphorylated along the proximal processes. The dorsal horn neurones probably contain 200 kD in a non-filamentous form but may lack the other subunits.
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  • 34
    ISSN: 1432-0878
    Keywords: Neurofilaments ; Phosphorylation ; Axon ; Immunocytochemistry ; Golden syrian hamster, Mesocricetus auratus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of phosphorylated and nonphosphorylated neurofilament epitopes was determined immunocytochemically in adjacent 2 μm-thick sections of sciatic nerve, ventral root and spinal cord. Staining was scored as either intense, moderate or absent and the proportion of labeled axons was calculated for each category. Nearly all sciatic nerve and ventral root axons were immunoreactive with both antibodies against phosphorylated and non-phosphorylated neurofilaments and there were no significant differences in the number of intensely- or moderately-labeled axons. Within the spinal cord however, while the majority of large caliber axons was stained with both antibodies, there was a significant number of small caliber axons which stained only with antibodies against phosphorylated neurofilaments. These results show that phosphorylated and nonphosphorylated neurofilaments are extensively codistributed in CNS and PNS axons, and that in the CNS, staining intensity for non-phosphorylated epitopes is less in the smaller axons.
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  • 35
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    Cell & tissue research 263 (1991), S. 271-284 
    ISSN: 1432-0878
    Keywords: Serotonin ; Immunocytochemistry ; Avian brain ; Hypothalamus ; Japanese quail, Coturnix coturnix japonica (Aves)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence and topographical localization of the serotoninergic system in the brain of the Japanese quail (Coturnix coturnix japonica) have been studied by means of peroxidase-anti-peroxidase immunocytochemistry. The perimeter, diameter, area, and shape factor of immunoreactive cells have been recorded and analyzed morphometrically for intra- and interspecies comparison. The data reported here confirm and extend results previously obtained in the brain of other avian species. Serotonin-immunoreactive neurons of the quail are mainly located in the hypothalamic paraventricular organ and adjacent areas, and in the brainstem where they form three separate groups. The first of these groups consists of small-sized neurons located in the ventro-rostral mesencephalon. The second group is composed of medium-sized neurons located in the dorsal mesencephalo-pontine region. The third group is also formed by medium-sized neurons, and is located ventrally in the ponto-medullary region. In the quail brain, serotoninergic neurons are not restricted to nuclei located in the vicinity of the midsagittal plane, but show some lateralization, especially in the brainstem. The organization of the different groups of immunoreactive neurons based on this topographical distribution and morphometric analysis has been compared with descriptions of the serotoninergic system in other birds. Serotonin-immunoreactive nerve fibers are widely distributed throughout the brain, but appear to be particularly abundant in regions involved in the control of reproductive activities, such as the septal region, the medial preoptic nucleus, the nucleus intercollicularis, and the external zone of the median eminence. The data reported here have allowed the drawing of a map of serotoninimmunoreactive structure.
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  • 36
    ISSN: 1432-0878
    Keywords: Met-enkephalin-Arg6-Gly7-Leu8 ; Immunocytochemistry ; Neuropeptides ; Allatostatins ; Neurosecretory cells ; Corpus cardiacum/corpus allatum complex ; Diploptera punctata, Calliphora vomitoria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neuronal circuits in the brain and retrocerebral complex of the cockroach Diploptera punctata have been mapped immunocytochemically with antisera directed against the extended enkephalin, Met-enkephalin-Arg6-Gly7-Leu8 (Met-8). The pathways link median and lateral neurosecretory cells with the corpus cardiacum/corpus allatum complex. In females, nerve fibres penetrate the corpora allata and varicosities or terminals, immunoreactive to Met-8, surround the glandular cells. Males differ in having almost no Met-8 immunoreactivity in the corpora allata. The corpora cardiaca of both males and females are richly supplied with Met-8 immunoreactive material, in particular in the ‘cap’ regions immediately adjacent to the corpora allata. A similarity in the amino-acid sequences of Met-8 and the C-terminus of the recently characterised allatostatins of D. punctata suggests that the pathways identified with the Met-8 antisera may be the same as those by which the allatostatins are transported from the brain to the corpus allatum. In comparative studies on the blowfly Calliphora vomitoria, similar neuronal pathways have been identified except that no sexual dimophism with respect to amounts of immunoreactive material within the corpus allatum has been observed. These results suggest a possible homology in the neuropeptide regulation of the gland.
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  • 37
    ISSN: 1432-0878
    Keywords: Vitamin D-1,25(OH)2D3-like immunoreactivity ; 1,25(OH)2D3 receptor-like immunoreactivity ; Immunocytochemistry ; Calcium ions ; Orchestia cavimana (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary For the first time, immunoreactivity to 1,25(OH)2D3-like molecules and their receptors has been investigated in a calcium transporting epithelium of a crustacean, Orchestia, using vertebrate antisera on ultrathin cryosections of posterior caeca previously fixed in glutaraldehyde, then postfixed in osmium tetroxide. Both immunoreactivities were located mainly in the nuclei of epithelial cells. Quantitative differences in 1,25(OH)2D3-like immunoreactivity were noticed from one stage of the molt cycle to another. These results, together with other data, contribute to evidence that immunoreactive 1,25(OH)2D3-like molecules may be involved in the regulatory processes of calcium metabolism in this terrestrial crustacean and suggest an involvement of these substances in the regulation of calcium movements in the posterior caeca.
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  • 38
    ISSN: 1432-0878
    Keywords: Pituitary gland, pars anterior (distalis) ; Gonadotrophic cells, gonadotropes ; Thyrotropes ; Immunocytochemistry ; Suncus murinus (Insectivora)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract According to recent immunocytochemical studies of anterior pituitary cells, it is obvious that the “one cell-one hormone” theory must be modified. Many pituitary morphologists have demonstrated that there are some cells that contain two hormones. In this study, we demonstrate by means of immuno-electronmicroscopy the co-existence of gonadotrophins (FSH and LH) and thyrotrophin (TSH) in the same anterior pituitary cells of the musk shrew. These cells were remarkably altered in their ultrastructural features by either gonadectomy or thyroidectomy. Double labeling for gonadotrophins and thyrotrophin was present not only in the same cells but also in the same secretory granules. Our ability to demonstrate co-existence of gonadotrophins and thyrotrophin in the same cell may be due to our selection of fixative and embedding media for electron-microscopic immunocytochemistry. Our conclusion that gonadotrophins and thyrotrophin are produced in a single cell type of the anterior pituitary gland in the musk shrew, i.e., thyrogonadotrophs, suggests the need to consider a modification of the classic scheme for classification of anterior pituitary cells.
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  • 39
    ISSN: 1432-0878
    Keywords: Opioid peptides ; Met-enkephalin ; Leuenkephalin ; Immunocytochemistry ; Nervous system, central ; Helix pomatia (Mollusca) ; Lymnaea stagnalis (Mollusca) ; Aplysia californica (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution and morphology of met-enkephalin-like immunoreactive (MetLI) and leu-enkephalin-like immunoreactive (LeuLI) neurons were investigated in the central nervous system of three gastropod species, Helix pomatia, Lymnaea stagnalis and Aplysia californica. Differences between the three species were observed in (1) the characteristics of immunostaining with antibodies to met-enkephalin and leu-enkephalin antibodies; (2) the number of immunostained neurons; (3) the projection areas of imunostained elements; (4) the specificity of immunostaining. Differences in the appearance of MetLI and LeuLI neurons were apparent: in Aplysia, both MetLI and LeuLI neurons could be observed, whereas in Lymnaea LeuLI was only found in fibers; only MetLI neurons occurred in Helix. According to an absorbtion control specificity test, a part of the LeuLI seen in the neuropil of Aplysia ganglia did not represent authentic leu-enkephalin. In Helix pomatia, significantly more MetLI neurons were present than in the CNS of Lymnaea and Aplysia; the majority of these neurons were concentrated in the cerebral ganglia and were small-size (12–25 μm) interneurons. In addition to central and peripheral projections observed in the three species, the connective tissue sheath around the ganglia and peripheral nerves contained MetLI varicose axons only in Helix, where a possible neurohormonal role could be attributed to these substance. The mapping and detailed chemical-neuroanatomical description of enkephalin-immunoreactive elements may furnish a chemical-neuroanatomical background to facilitate further neurophysiologic and pharmacologic analysis of enkephalinergic mechanisms in the gastropod CNS.
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  • 40
    ISSN: 1432-0878
    Keywords: Somatotrophs ; Somatotroph granules ; Growth hormone ; Immunocytochemistry ; Developments ontogenic ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunocytochemical methods were used to gain information about the embryonic development of chicken somatotrophs before and after hatching. To localize growth hormone, anterior pituitary sections were incubated with growth-hormone antibody, and then an indirect peroxidase method was used for light microscopy and an immunogold method for electron microscopy. The earliest evidence of embryonic somatotrophs was seen at 12 days. At this stage somatotrophs were sparse (0.2% of parenchymal cells) and their granules were pleomorphic with elongated ovoid and lozenge shapes predominating. Few of the immunogold-labeled somatotroph granules of the embryo were spherical until 15 days after fertilization. At 18 days, most of the granules were spherical (their shape in the adult chicken). During the six days between the 15-day-old embryo and the 1-day-old chick, the number of gold particles per granule section approximately doubled suggesting an increase in growth hormone content of the granules. This rise was the result of increases in the size of the granule sections and in the concentration of gold particles in the sections. During the embryonic period of 12–20 days, somatotrophs were not more than 3.6% of the anterior pituitary cell population. During the following two days, between the 20-day-old embryo and the 1-day-old chick, the percentage of somatotrophs in the pituitary parenchymal cell population rose rapidly from 3.6% to 20.7% and then increased slowly to 24.6% during the period of 1–5 days after hatching. Both the sharp percentage rise in somatotrophs (20-day-old embryo to 1-day-old chick) and the rise in growth hormone content of the granules (15-day-old embryo to 1-day-old chick) suggested by gold-particle counts occur close to the time of hatching. These morphological changes may reflect an increased synthesis of growth hormone that is responsible for the rise in plasma growth-hormone concentration that begins about the same time and is especially abrupt two days later (1–3 days after hatching).
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  • 41
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    Cell & tissue research 264 (1991), S. 23-32 
    ISSN: 1432-0878
    Keywords: Male urethra ; Urethral epithelium ; Immunocytochemistry ; Ultrastructure ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distal segment of the human male urethra, in particular the fossa navicularis, was studied with light- and electron microscopy as well as by means of histochemical and immunocytochemical methods. The fossa navicularis of the urethra contains a circumscribed zone of extremely thick, non-keratinized stratified squamous epithelium composed of cells containing a large amount of glycogen. These cells lack acid phosphatase activity and lysozyme-like immunoreactivity, both of which can be demonstrated to varying extents in the other zones of the distal male urethra. These glycogen-rich cells are considered to be the substrate for an endogenous flora of lactobacteria, whereas the acid-phosphatase activity and the lysozyme-like immunoreactivity indicate the presence of macrophages and the secretion of bactericidal agents at the epithelial surface. These observations suggest that the different zones with heterogeneous properties in the distal male urethra probably represent a defense system against the invasion of pathogenic microorganisms. Moreover, the glycogen-rich zone, which resembles the glycogen-rich epithelium of the vagina, is estrogen-dependent. This is demonstrated in cases of sex reversal in which after long-lasting estrogen treatment the glycogen-rich zone becomes extremely extended by displacement of the neighbouring epithelium.
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  • 42
    ISSN: 1432-0878
    Keywords: Enkephalins ; Tegmentum mesencephali ; Tectum opticum ; Immunocytochemistry ; Retrograde labeling ; Cobalt-lysine ; Salmo gairdneri, Salmo salar (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemistry using antibodies against Met-enkephalin and Leu-enkephalin has demonstrated a group of large enkephalin-immunoreactive neurons in the nucleus of the rostral mesencephalic tegmentum (mRMT) of two teleost fish, Salmo gairdneri and Salmo salar. Injections of cobalt-lysine in the medial optic tectum retrogradely labeled the above group of tegmental neurons. Tegmental neurons were labeled only ipsilaterally to the injection site. This indicates that enkephalinergic neurons in the nRMT project to the optic tectum, and that at least some of the enkephalinergic axons observed in the optic tectum belong to a tegmento-tectal pathway. Comparable enkephalinergic pathways have been described in reptiles and birds, where pretectal-mesencephalic nuclei contribute to the enkephalin-containing fibers that project to the optic tectum.
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  • 43
    ISSN: 1432-0878
    Keywords: Pancreas, endocrine ; Larval development ; Serial thin/semithin sections ; Immunocytochemistry ; Rana temporaria (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The pancreatic endocrine component was studied at different stages of development in the tadpoles of Rana temporaria. The material was embedded in Epon, and serial semithin and thin sections were made in order to correlate ultrastructural features and tinctorial traits of the endocrine cells. Serial semithin sections were also stained with the peroxidase-antiperoxidase immunocytochemical method and with silver impregnations for argyrophilia and argentaffinity. In early larvae (legless tadpoles), A and B cells are present. Both can be found within ducts and exocrine tissue or, more frequently, in cellular clusters among the ducts and acini. These primitive islets are solid structures, surrounded but not penetrated by capillaries. Mitoses were observed in A and B cells. In the following phase (tadpoles with hindlegs), D and pancreatic polypeptide-immunoreactive cells are also present, as well as numerous endocrine cells scattered among exocrine tissue. There is also a change in the vascular-insular pattern: capillaries not only surround but also penetrate the endocrine group. The structure of the endocrine pancreas in older tadpoles is similar. Tinctorial traits and ultrastructural features of endocrine cells are described, and the origin of primitive islets is discussed.
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  • 44
    ISSN: 1432-0878
    Keywords: Glutamate ; Nervous system, insect ; Ganglia, invertebrate ; Immunocytochemistry ; Schistocerca gregaria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of glutamate-like immunore-activity in the thoracic and abdominal ganglia of the locust was studied using two polyclonal antibodies against glutamate. Because glutamate is a precursor of the inhibitory transmitter γ-amino butyric acid (GABA) the distribution of immunostaining by antibodies against glutamate and GABA was closely compared in adjacent serial sections. When the antibodies were used at optimal dilutions there was no overlap in the distribution of immunostaining for glutamate and GABA. In the pro- and mesothoracic ganglia 360–400 somata are immunoreactive for glutamate, while in the metathoracic ganglion about 600 somata were stained. These range in diameter from 10–100 μm in diameter and include the majority of the large somata in these ganglia. Bundles of primary neurites emerging from these large somata can be traced through the neuropile. Most of the bundles correspond to the known paths of motor neurone primary neurites. In addition the ‘T’-tract is also immunolabelled. The free abdominal ganglia each contain 80–100 somata ranging in size from 10–45 μm while the terminal ganglion contains about 250 somata, 10–60 μm in diameter.
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  • 45
    ISSN: 1432-0878
    Keywords: Gap junction ; Cell junction ; Immunocytochemistry ; Biochemistry ; Heliothis virescens (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Subcellular fractions enriched in gap junctions with an ultrastructure similar to those in intact insect tissue have been obtained by extracting crude membranes from the tobacco budworm Heliothis virescens (Lepidoptera: Noctuidae) with 2.5 mM NaOH. n-Octyl-β-d-glucopyranoside (OG) was used to further purify integral membrane proteins in the NaOH-extracted fractions. A polyclonal antibody (R16) is described that specifically labels nonextracted and NaOH-extracted gap junctions in cell fractions by electron microscope immunocytochemistry. R16 immunostaining of sectioned Heliothis testis at the light-microscope level yields a pattern of immunoreactivity consistent with the distribution of gap junctions in the tissue. R16 identifies a 40-kDa protein as a candidate gap junction protein on immunoblots of crude membrane, NaOH-extracted and NaOH/OG-extracted fractions.
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  • 46
    ISSN: 1432-0878
    Keywords: IAPP (islet amyloid polypeptide) ; Endocrine pancreas ; In situ hybridization ; Immunocytochemistry ; Somatostatin ; Insulin ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The expression of the islet amyloid polypeptide (IAPP) gene within the endocrine pancreas and its correlation with insular neuroendocrine peptide localization were investigated in the rat. In situ hybridization with a 35S-labelled IAPP-mRNA specific oligonucleotide probe was combined with immunocytochemistry. In situ hybridization alone showed strong autoradiographic labelling of the pancreatic islets. In situ hybridization combined with immunocytochemistry for IAPP, revealed labelling of the IAPP-immunoreactive cells. However, when in situ hybridization was combined with immunocytochemistry for proinsulin, we noted a lack of proinsulin immunoreactivity in some peripherally located autoradiographically labelled islet cells. Furthermore, combination of in situ hybridization and immunocytochemistry for somatostatin showed autoradiographic labelling of somatostatin cells to a varying degree. This was further confirmed by showing cellular co-localization of IAPP and somatostatin by immunocytochemical double staining. We conclude that IAPP is mainly synthesized in insulin cells. Additionally, a subpopulation of the somatostatin cells is capable of IAPP synthesis. This may account for the relatively small reduction in the content of IAPP-mRNA in islets compared to the marked reduction of insulin mRNA after streptozotocin-induced diabetes in rats as previously reported.
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  • 47
    ISSN: 1432-0878
    Keywords: Parvalbumin ; Peripheral vestibular system ; Crista ampullaris ; Utricle ; Immunocytochemistry ; Mouse (CBA/C57) ; Rat (Wistar) ; Guinea pig (BFA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The cellular distribution of parvalbumin immunoreactivity in the vestibular peripheral system of mouse, rat, and guinea pig was investigated by light and electron microscopy. Parvalbumin was found in all neurons of the vestibular ganglia of these species but in the sensory epithelia immunoreactivity was restricted to type I hair cells localized exclusively in the central areas. The very intense staining pattern was similar in the cristae ampullares and utricles of all three species but a faint immunoreaction was also detectable in sensory cells of peripheral areas of rat cristae. The parvalbumin-immunoreactive type I sensory cells are connected by nerve fibres of the calyx unit type which are known selectively to contain calretinin.
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  • 48
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Neuropeptide Y ; Gonadotropin-releasing hormone ; Reproductive function ; Domestic chicken, Gallus domesticus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Nerve fibers and perikarya containing gonadotropin-releasing hormone (GnRH-like) immunoreactivity were investigated in the brain of the three-week-old chick, Gallus domesticus using the technique of immunocytochemistry. Six major groups of perikarya were found to include the olfactory bulb, olfactory tubercle/lobus parolfactorius, nucleus accumbens, septal preoptic hypothalamic region (three sub-nuclei), lateral anterior thalamic nucleus and in and about the oculomotor complex. The immunostaining was unusual in the latter group, suggesting that the neurons may contain a GnRH-II like material. Immunoreactive fibers for GnRH were found throughout the entire brain extending from the olfactory bulbs to the caudal brainstem. Two anatomical areas, not emphasized in the past literature, which had distinct GnRH-like immunoreactivity, included the lateral anterior thalamic nucleus and the preoptic recess. The former included a group of GnRH perikarya that is also known to be a retino-recipient area while the latter contained neuronal terminals some of which appeared to be contacting the cerebrospinal fluid of the preoptic recess. An attempt was made to list all anatomical structures that contained or were juxta-positioned to sites that displayed immunoreactive perikarya and fibers including circumventricular organs.
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  • 49
    ISSN: 1432-0878
    Keywords: Oviduct ; Epithelial cell ; Glycoproteins ; Immunocytochemistry ; Cow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The immunocytochemical localization of bovine oviduct-specific glycoproteins was investigated by light and electron microscopy. Using monoclonal antibodies (MAbs) specific for bovine oviductal glycoproteins, 3 regions (fimbriae, ampulla, and isthmus) of the epithelium in the bovine oviduct we studied during the follicular and luteal phases. The MAbs reacted specifically with the oviductal epithelial cells. Intense labeling was observed in the ampullar and fimbrial epithelia of cows at the follicular phase, but the reactions were weaker at the luteal phase. In the isthmus, the immunohistochemical reaction was faint during both follicular and luteal phases. At the ultrastructural level, the MAbs bound selectively to putative secretory granules of nonciliated cells in the ampulla and fimbriae, but not in the isthmus. These results suggest that there are cyclic changes and regional differences in the production of glycoproteins in the bovine oviduct.
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  • 50
    ISSN: 1432-0878
    Keywords: Pineal organ ; Retina ; Photoreceptors ; Photopigment ; Immunocytochemistry ; HPLC ; Autoradiography ; Mouse (C57BL)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The aim of the present study was to characterize the rod-opsin immunoreaction in the mammalian pineal organ. Pigmented mice (strain C57BL) were selected as the animal model. Immunocytochemical investigations involving the use of highly specific polyclonal and monoclonal antibodies against bovine rod-opsin (the apoprotein of the photopigment rhodopsin) showed that approximately 25% of all pinealocytes were rod-opsin immunoreactive. Immunoblotting techniques revealed three protein bands of approximately 40, 75, and 110 kDa; these were detected by the monoclonal antibody and the polyclonal antiserum in retinal and pineal extracts. These protein bands presumably represented the monomeric, dimeric and trimeric forms of rod-opsin. The amount of rod-opsin in retina and pineal organ was quantified by means of an enzyme-linked immunosorbent assay. This yielded 570±30 pmoles rod-opsin per eye and 0.3±0.05 pmoles rod-opsin per pineal organ. High pressure liquid chromatography analysis of whole eye extracts demonstrated the chromophoric group of the photopigment rhodopsin, 11-cis retinal, and its isomer, all-trans-retinal. A shift from 11-cis retinal to all-trans-retinal was found upon light adaptation. No retinals were detected in the pineal organ. Autoradiographic investigations showed that 3H-retinol, intraperitoneally injected into the animals, was incorporated into the outer and inner segments of retinal photoreceptors, but not into the pineal organ. It is concluded that the mouse pineal organ contains the authentic apoprotein of rhodopsin but that it lacks retinal derivatives as essential components of all known vertebrate photopigments. Consequently, the “photoreceptor-specific” proteins of the mammalian pineal organ are not involved in photoreception and phototransduction, but may serve other functions to be explored in future studies.
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  • 51
    ISSN: 1432-0878
    Keywords: Rhabdomeres ; Cytoskeleton ; Actin ; Immunocytochemistry ; Membrane shedding ; Leptotarsus spp. (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Rhabdomeres of tipulid flies lose membrane during turnover from a ‘shedding zone’ composed of microvillar tips. These distal domains lack intramicrovillar cytoskeletons and appear to be empty sacs of membrane. Recent concerns about the role of ninaC mechano-enzymes in the architecture of dipteran rhabodomeral microvilli and the dynamic role that they may play in the creation of shedding zones demand an examination of the distribution of actin in tipulid rhabdomeres. We compared rhabdomeres from tipulid retinae incubated before fixation for immunocytochemistry in a buffer without additives and a stabilising buffer that contained a cocktail of cysteine protease inhibitors; both were challenged by an anti-actin antibody for immunogold labelling after embedding in LR White Resin. Shedding zones thus processed collapse to structureless detritus. Stabilised and unstabilised shedding zones were immunonegative to anti-actin. To ensure that the negative results were not consequent upon conformational changes generated by the processing protocol, we examined microvilli of degenerating rhabdomeres of the Drosophila light-dependent retinal degeneration mutant rdgB KS222 (which separate and collapse without creating a shedding zone) and found the detritus they generate to be immunopositive to anti-actin. Stabilised and unstabilised regions of basal regions of tipulid rhabdomeres were equally immunopositive. We infer that (a) actin is absent from shedding zones; (b) actin is not degraded by microvillar cysteine proteases. The implications of these conclusions are discussed in relation to some functional models of arthropod photoreceptor microvilli.
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  • 52
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    Cell & tissue research 265 (1991), S. 485-492 
    ISSN: 1432-0878
    Keywords: ACTH ; Brain ; Hypothalamus ; Hypophysectomy ; Cortisol ; Metopirone ; Immunocytochemistry ; Pituitary ; Corticotropin-releasing factor ; Anguilla anguilla (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An ACTH-like peptidergic system was demonstrated in the brain of three teleost species by immunocytochemistry. In order to investigate the origin of brain ACTH and factors modulating its synthesis, similar techniques were applied to the brain of eels (1) submitted to hypothysectomy in order to suppress pituitary ACTH and plasma cortisol, (2) injected with cortisol to inhibit pituitary ACTH synthesis and release, and (3) injected with metopirone to block cortisol synthesis and stimulate ACTH synthesis and release. Hypophysectomized eels showed a normal distribution of immunoreactive perikarya in the ventral hypothalamus and fibers in the brain, suggesting that brain ACTH does not arise from the pituitary. In cortisol-treated eels immunostaining was markedly reduced in brain perikarya and pituitary corticotropes, suggesting a reduced synthesis. In metopirone-injected eels, one third of the animals showed an increased immunostaining in perikarya and a dense network of immunoreactive fibers, suggesting that ACTH synthesis was increased. Brain ACTH was not affected in other animals. Pituitary corticotropes were rapidly degranulated. Responses of ACTH in the brain and pituitary occur independently when cortisol synthesis is inhibited. These responses are compared to those of the corticotropin-releasing factor system in the same eels.
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  • 53
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    Cell & tissue research 265 (1991), S. 527-534 
    ISSN: 1432-0878
    Keywords: Autonomic nervous system ganglia ; Enteric nervous system ; Intestine, large ; Tissue culture ; Immunocytochemistry ; Peptides ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ganglia of the myenteric plexus from the newborn guinea-pig, isolated by microdissection, were dissociated by a combination of enzymatic and mechanical methods. The neurones and glial cells in the resulting cell suspension were cultured for up to 21 days in vitro. The growth of the enteric ganglion cells in serum-free, hormone-supplemented (N1) medium and in serum-supplemented medium containing a mitotic inhibitor was compared over a period of 14 days in vitro. Enteric neurones were outnumbered by glia in both culture media, although glial cell proliferation was inhibited in both media compared with that in serum-supplemented medium without mitotic inhibitors. Glial cell numbers appeared to decline in serum-free medium after the first week in vitro. Neurites tended to be more varicose in the serum-free medium, and the morphology of the enteric glial cells also differed markedly in the two media. This is the first report of the dissociation and subsequent culture of myenteric ganglia that had previously been completely isolated from the remainder of the gut wall.
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  • 54
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    Cell & tissue research 265 (1991), S. 517-525 
    ISSN: 1432-0878
    Keywords: Spermiogenesis ; Spermatids ; DNA ; Immunocytochemistry ; Electron microscopy ; Bovine ; Mouse ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary DNA distribution in mouse, rabbit and bull spermatids was analyzed by electron microscopy, after using a Feulgen-like HCl-osmium ammine procedure, and after immunocytochemistry with anti-DNA antibodies. In addition, nucleic acids were visualized with the intercalating dye ethidium bromide and phosphotungstic acid. The parts of DNA displaying a beta helix configuration (possibly A-T rich parts) were identified by epifluorescence microscopy after staining with Hoechst 33258. In all 3 species, young spermatid nuclei were seen to have large areas poor in DNA, as well as DNA-rich areas, which were mostly concentrated into a peripheral layer close to the acrosome and into one or several masses, displaying species-specific locations. These DNA-rich areas were stained with Hoechst 33258. Elongating spermatid nuclei contained homogeneously distributed DNA, and this was evident following both immunocytochemistry and nucleic acid histochemistry in all 3 species. However, the distribution appeared more heterogeneous after the Feulgen-like procedure, and was accompanied by a disappearance of Hoechst-fluorescence. In fully elongated spermatids, all nuclear areas stained with Hoechst 33258, while the 3 other techniques labeled either all or species-specific parts of the condensed chromatin. The reasons for these variable reactions are discussed in terms of technique specificities, DNA configuration and nucleoprotein moiety replacements.
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  • 55
    ISSN: 1432-0878
    Keywords: Retina ; Melatonin ; Rhythmic biosynthesis ; Immunocytochemistry ; Pike, Esox lucius (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The levels of melatonin and the activities of two enzymes of the melatonin biosynthetic pathway, serotonin N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT), were measured throughout the light-dark cycle in the retina of a teleost fish, the pike. HIOMT activity did not display significant variations, whereas NAT activity and melatonin content showed a daily rhythm, high levels occurring during the night. The profiles of the latter two rhythms did not closely match one another and differed from those previously described in the pineal organ of the same species. These results are discussed with respect to a possible paracrine role of retinal melatonin. Melatonin-like immunoreactivity was found in the photoreceptor cell layer and in the Müller cells of the inner nuclear layer. The intensity of the melatonin-like immunoreactivity varied throughout the 24 h light-dark cycle, in good correlation with the variations in the melatonin level as measured by radioimmunoassay.
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  • 56
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    Cell & tissue research 265 (1991), S. 611-614 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Met-enkephalin ; Merkel cells ; Dense-core granules ; Mouse (ICR)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The electron-microscopic immunogold method was applied to Merkel cells of adult mice to demonstrate the subcellular localization of met-enkephalin-like immunoreactivity. Post-embedding incubation with metenkephalin antisera showed that the gold particles were associated with the dense-core granules of the Merkel cells. The majority, but not all, of the dense-core granules were strongly labelled. Osmication caused a significant reduction in the number of gold particles on these granules. The nerve terminal associated with the Merkel cell did not show met-enkephalin-like immunoreactivity. To the best of our knowledge, this is the first report of the ultrastructural localization of a positive met-enkephalin immunoreactivity in the dense-core granules of Merkel cells in mice.
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  • 57
    ISSN: 1432-0878
    Keywords: Leukocytes ; T-Lymphocytes ; B-Lymphocytes ; Development, ontogenetic ; Digestive tract ; Immunocytochemistry ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In the present study, we characterized intra-epithelial leukocytes in the digestive tract of chickens during postnatal development. Their phenotype was characterized by monoclonal antibodies in cryostat sections and the numbers of the different cell-types were counted in the epithelium of the esophagus, proventriculus, duodenum, jejunum, cecum, and colon. All intra-epithelial leukocytes bore the leukocyte-common antigen CD45; 35% were T lymphocytes, and 50% bore a B-cell marker. However, no immunoglobulin-bearing cells were detected in the epithelium. Monocytes and macrophages were found only in the epithelium of the esophagus. A remaining population of non-B, non-T, non-monocyte cells (15%) was present in all parts of the digestive tract. The number of intra-epithelial leukocytes was greatest in the duodenum and jejunum, and decreased in the proximal part of the cecum and in the colon. Intra-epithelial leukocytes were only sporadically detected in the proventriculus. The total number of intra-epithelial leukocytes increased until 8 weeks after hatching and then decreased at 18 months. In the esophagus, the total number of intra-epithelial leukocytes changed little during aging. We found that the intra-epithelial leukocytes of chickens and rodents are distinct in that chicken intra-epithelial leukocytes comprise a cell population that bears a B-cell antigen but that lacks surface immunoglobulins.
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  • 58
    ISSN: 1432-0878
    Keywords: Kallikrein ; Prolactin ; Estrogen ; Haloperidol ; Anterior pituitary ; Immunocytochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Glandular kallikrein (GK, a trypsin-like serine protease) exhibits estrogen induction and dopamine repression in rat pituitary lactotrophs. Steroid induction may reflect primary actions to increase selectively the synthesis of specific proteins, or may be part of broad cellular responses secondary to steroid-induced phenotype transitions. This study examined the cellular mechanisms underlying estrogen and dopaminergic control of lactotroph GK using a quantified immunocytochemical approach. Pituitaries from ovariectomized rats exhibited little GK staining. Estradiol treatment for 10 days produced dose-dependent increases in pituitary mass, the percentage of lactotrophs (indicating lactotroph proliferation) and the percentage of GK-positive cells. Also, GK staining intensity was dependent upon estradiol dose, increasing 4-fold between 5 μg and 50 μg/48 h. Dopamine receptor blockade with haloperidol (2.5 mg/kg/24 h) elicited weak GK immunostaining in 46% of the lactotrophs in the absence of estradiol, and markedly potentiated GK staining intensity elicited with low but not high doses of estradiol. The results suggest that GK induction is a primary estrogen effect, and is not secondary to a phenotype transition: the induction is enhanced by estrogen-induced lactotroph proliferation. Dopaminergic systems strongly inhibit GK induction by low estradiol levels. This dopaminergic modulation may shift the induction of lactotroph GK to physiological events associated with high estradiol levels or low dopaminergic tone.
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  • 59
    ISSN: 1432-0878
    Keywords: Nervous system, central ; Dopamine ; Tyrosine hydroxylase ; Catecholamines ; Immunocytochemistry ; Helix pomatia (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution and characterization of dopamine-containing neurons are described in the different ganglia of the central nervous system of Helix on the basis of the distribution of tyrosine hydroxylase immunoreactive (TH-ir) and dopamine immunoreactive (DA-ir) neurons. Both TH-ir and DA-ir cell bodies of small diameter (10–25 μm) can be observed in the buccal, cerebral and pedal ganglia, dominantly on their ventral surface, and concentrated in small groups close to the origin of the peripheral nerves. The viscero-parietal-pleural ganglion complex is free of immunoreactive cell bodies but contains a dense fiber system. The largest number of TH-ir and DA-ir neurons can be detected in the pedal, and cerebral ganglia. The average number of TH-ir and DA-ir neurons significantly differs but all the identifiable groups of TH-ir neurons also show DA-immunoreactivity. Therefore, we consider the TH-ir neurons in those groups as being DA-containing neurons. The amounts of DA in the different ganglia assayed by high performance liquid chromatography correspond to the distribution and number of TH-ir and DA-ir neurons in the different ganglia. The axon processes of the labeled small-diameter neurons send thin proximal branches toward the cell body layer but only rarely surround cell bodics, whereas distally they give off numerous branches in the neuropil and then leave the ganglion through the peripheral nerves. In the cerebral ganglia, the analysis of the TH-ir pathways indicates that the largest groups of labeled neurons send their processes through the peripheral nerves in a topographic order. These results furnish morphological evidence that DA-containing neurons of Helix pomatia have both central and peripheral roles in neuronal regulation.
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  • 60
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    Cell & tissue research 274 (1993), S. 579-585 
    ISSN: 1432-0878
    Keywords: Thyroxine ; Pituitary gland, pars distalis ; Immunocytochemistry ; Ultrastructure ; Mouse (Snell dwarf)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The effects of dietary thyroxine on the immunoreactivity of cells in the pars distalis of the adenohypophysis in dwarf (dw/dw) mice were determined by ultrastructural immunocytochemistry. In nontreated dwarfs only adrenocorticotropic hormone (ACTH) cells and luteinizing hormone (LH) cells showed positive reactions to their respective antibodies, whereas no cells showed immunoreactivity to antibodies to growth hormone (GH), thyroid-stimulating hormone (TSH), or prolactin (Prl). In dwarfs supplemented postnatally with dietary thyroxine for 9 wks, the treatment failed to produced immunoreactive GH, TSH or Prl cells. However, LH cells became more prominent and fully developed, with denser concentrations of immunoreactive particles overlying the secretory granules than occurred in nontreated dwarfs. In thyroxine-treated dwarfs, ACTH cells were similar in ultrastructural features and immunoreactivity to those in nontreated dwarfs.
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  • 61
    ISSN: 1432-0878
    Keywords: Pituitary ; Pars distalis ; Growth hormone ; Prolactin ; Hormonal specificity ; Immunocytochemistry ; Immunoblot technique ; Amphibians (Urodela, Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract An antiserum was prepared against the recently purified bullfrog (bf) growth hormone (GH); it was applied to sections of brain and pituitary of three urodele (Ambystoma, Pleurodeles and Cynops) and three anuran (Xenopus, Bufo vulgaris and B. japonicus) species. No immunostaining was obtained in the urodele pituitary, being consistent with the results of immunoblot analysis of the pituitary homogenate. In the three anuran species, strong immunoreactivity was observed in GH cells that were concentrated in the posterodorsal region of the pars distalis. No GH-like immunoreactivity was detectable in the brain of any of the species. A comparison using adjacent sections stained with anti-bf prolactin (PRL) confirmed the anteroventral localization of PRL cells. Colocalization of GH and PRL was not apparent. These data suggest that the molecular structure of amphibian GHs is considerably different between anurans and urodeles. The antiserum used in the present work shows a high species specificity, recognizing only anuran GHs. In contrast anti-bfPRLlabeled PRL cells in all the amphibian species studied in the present work, suggesting that PRLs possess common amino acid sequences recognized by the anti-bfPRL.
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  • 62
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    Cell & tissue research 266 (1991), S. 247-258 
    ISSN: 1432-0878
    Keywords: Monoclonal antibodies ; Immunocytochemistry ; Olfactory epithelium ; Respiratory epithelium ; Antigens ; (Rat Wistar) ; (Mouse BALB/c)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Two panels of monoclonal antibodies have been generated, each panel having a distinct specificity for antigens located in the ciliary zone of either the olfactory or respiratory epithelium of rats. Tissue specificity was confirmed in enzyme-linked immunosorbent assays on membrane fractions from various tissues. During ontogeny, the expression of olfactory-specific antigens preceeds that of respiratory-specific antigens; this observation correlates with differences in the genesis of the respective cilia type and confirms that different molecular entities are recognized. A spatial segregation of immunoreactivity in the chemosensory epithelium was observed for one of the olfactory-specific monoclonal antibodies; negative zones were located in the dorsal recess of the nasal cavity and on the tips of the turbinates. Olfactory-specific antibodies reacted with distinct polypeptide bands on Western blots from olfactory ciliary preparations.
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  • 63
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    Cell & tissue research 266 (1991), S. 359-364 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Neuropeptide ; Diuretic hormone ; Neurosecretion ; Manduca sexta, Eurema nicippe (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Separate antisera were raised to the N- and C-terminal half of the diuretic hormone from Manduca sexta. Antisera against the two halves of this peptide recognized the same cells in M. sexta, and preabsorption of the antisera with the peptides used as antigens abolished the immunoreactivity, confirming their specificity. The antisera reacted with two median neurosecretory cells on each side of the protocerebral groove in larvae, and with a group of about 80 small median neurosecretory cells in the adult, as well as their axons to, and their axon terminals in, the corpora cardiaca. During the early pupal stages, small cells, which are possibly derived from a common neuroblast, differentiate into immunoreactive neurosecretory cells, which explains the large increase in cell numbers in the adult. In the sleepy sulphur butterfly, Eurema nicippe, homologous median neurosecretory cells in the adult were immunoreactive with both antisera.
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  • 64
    ISSN: 1432-0878
    Keywords: Pigment-dispersing hormone ; Orthopteroid insects ; Immunocytochemistry ; Insect brain ; Periplaneta americana, Schistocerca gregaria, Teleogryllus commodus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In a comparative study, the anatomy of neurons immunoreactive with an antiserum against the crustacean β-pigment-dispersing hormone was investigated in the brain of several orthopteroid insects including locusts, crickets, a cockroach, and a phasmid. In all species studied, three groups of neurons with somata in the optic lobes show pigment-dispersing hormone-like immunoreactivity. Additionally, in most species, the tritocerebrum exhibits weak immunoreactive staining originating from ascending fibers, tritocerebral cells, or neurons in the inferior protocerebrum. Two of the three cell groups in the optic lobe have somata at the dorsal and ventral posterior edge of the lamina. These neurons have dense ramifications in the lamina with processes extending into the first optic chiasma and into distal layers of the medulla. Pigment-dispersing hormone-immunoreactive neurons of the third group have somata near the anterior proximal margin of the medulla. These neurons were reconstructed in Schistocerca gregaria, Locusta migratoria, Teleogryllus commodus, Periplaneta americana, and Extatosoma tiaratum. The neurons have wide and divergent arborizations in the medulla, in the lamina, and in several regions of the midbrain, including the superior and inferior lateral protocerebrum and areas between the pedunculi and α-lobes of the mushroom bodies. Species-specific differences were found in this third cell group with regard to the number of immunoreactive cells, midbrain arborizations, and contralateral projections, which are especially prominent in the cockroach and virtually absent in crickets. The unusual branching patterns and the special neurochemical phenotype suggest a particular physiological role of these neurons. Their possible function as circadian pacemakers is discussed.
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  • 65
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    Cell & tissue research 266 (1991), S. 385-389 
    ISSN: 1432-0878
    Keywords: Amniotic epithelium ; Cytoskeleton ; Filaments ; α-Actinin ; Ezrin ; Immunocytochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The amniotic epithelium constitutes a diffusion barrier controlling the passage of solutes and water between the aminotic cavity and maternal circulation. With the present immunocytochemical approach, we have shown that several major components of the cytoskeleton, i.e., actin, α-actinin, spectrin and ezrin, are preferentially associated with the apical and lateral cell surfaces of the human amniotic epithelium. Keratins are distributed throughout the entire cytoplasm, whereas vimentin mainly forms a perinuclear scaffold. These findings indicate a role of the various components of the cytoskeleton in the structural integrity and modulation of cell shape and junctional permeability.
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  • 66
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    Cell & tissue research 271 (1993), S. 177-179 
    ISSN: 1432-0878
    Keywords: Bone resorption ; Osteoclast ; Cysteine proteinase ; Immunocytochemistry ; Rats (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cysteine-proteinases such as cathepsin B and G were localized in rat osteoclasts, by an indirect protein A-immunogold labeling technique, on post-embedded ultrathin sections. In osteoclasts, specific immunogold labeling of both anti-cathepsin B and G was localized in Golgi vesicles, lysosomes, pale vacuoles of various sizes, and the extracellular canals of ruffled borders; no immunoreactivity was seen in the cytoplasmic matrix, mitchondria, cisterns of the rough endoplasmic reticulum, or nuclei. The presence of immunolabeling of cathepsins in osteoclasts and in the subosteoclastic compartment suggests that these enzymes are involved in the extracellular degradation of collagen and other noncollagenous bone matrix proteins.
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  • 67
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    Cell & tissue research 271 (1993), S. 279-288 
    ISSN: 1432-0878
    Keywords: Nervous system, central ; Ganglia, invertebrate ; Immunocytochemistry ; GABA (γ-aminobutyric acid) ; Glutamate decarboxylase (GAD) ; Eriphia spinifrons (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have used specific antisera against protein-conjugated γ-aminobutyric acid (GABA) and rat-brain glutamic acid decarboxylase (GAD) in immunocytochemical preparations to study the distribution of putatively GABAergic neurons in the fused thoracic ganglion of the crab Eriphia spinifrons. In the thoracic neuromeres, about 2000 neurons with somata arranged in clusters or located singly in the cell cortex exhibited both GABA-like and GAD-like immunoreactivity. In addition, more than a hundred cells showed only GABA-like immunoreactivity. Fibrous immunoreactive staining to GAD and GABA was distributed throughout the neuropil of the thoracic ganglion, and several fiber tracts contained immunoreactive processes. Sets of serially homologous neurons exhibited GABA-like and GAD-like immunoreactivity in the thoracic neuromeres. Especially prominent were one medial and four ventro-lateral clusters of somata, together with thirteen individually recognized cells in each neuromere. Six of these cells in the ventro-medial cell cortex may be the somata of inhibitory motoneurons. The leg nerves contained three immunoreactive fibers, corresponding to the previously described common inhibitory motoneuron and the two specific inhibitors. The results present further evidence for GABA being the neurotransmitter of all inhibitory leg motorneurons, and suggest its presence and role as a neurotransmitter in a considerable number of interneurons in the thoracic ganglion of the crab.
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  • 68
    ISSN: 1432-0878
    Keywords: Somatolactin ; Pituitary ; Gene expression ; In situ hybridization ; Immunocytochemistry ; Protein A-gold ; Oncorhynchus mykiss (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The gene expression and intracellular localization of somatolactin (SL), a putative pituitary hormone structurally related to both growth hormone and prolactin, were investigated in the pituitary of rainbow trout, Oncorhynchus mykiss. Using an in situ hybridization technique, we demonstrated the gene expression of the SL molecule in cells bordering the neurohypophysial tissue in the pars intermedia. These cells were identified immunocytochemically as SL-cells on the adjacent section. Electron-microscopic immunocytochemistry by means of the protein A-gold technique, also revealed that the SL-immunoreactivity was located mostly on the secretory granules in SL-cells. Our findings clearly indicate that SL is biosynthesized and stored in the granules in these cells.
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  • 69
    ISSN: 1432-0878
    Keywords: Colon ; Immunocytochemistry ; Intestine large ; Ionic regulation ; Na+ ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have used polyclonal antibodies generated against purified bovine renal amiloride-sensitive Na+ channels to localize amiloride-sensitive Na+ channels within the lower intestine (colon and coprodeum) of the hen. These antibodies cross-reacted with two polypeptides exhibiting Mr's of 235 and 150 kDa on immunoblots of detergent-solubilized apical membrane fractions from both the colon and coprodeum. The apparent molecular masses of theses polypeptides are in agreement with the Mr's of 2 of the subunits of the renal high amiloride-affintiy Na+ channel, namely the α and the β(=amiloride binding) subunits. The cellular distribution of Na+ channels was determined by immunoperoxidase and indirect immunofluorescence cytochemical techniques. The apical (luminal) membrane and cytoplasm of villar principal cells in both colon and coprodeum exhibited immunoreactivity, whereas goblet cells were nagative. Both principal and goblet cells of the crypts were also negative. We conclude that the amiloride-sensitive Na+ channels are localized to the principal cells of the intestinal villi and that these cells are responsible for intestinal Na+ absorption.
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  • 70
    ISSN: 1432-0878
    Keywords: Pituitary ; Galanin ; Neuromedin-U ; Corticotropes ; Immunocytochemistry ; Electron microscopy ; Plasticity ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localization of galanin in rat lactotropes and human corticotropes is well established. Neuromedin U immunoreactivity is present in rat corticotropes but radioimmunoassay of thyroid-manipulated rat pituitaries has also linked it to the thyroid axis. We found galanin immunoreactivity in some rat corticotropes, so we have re-examined rat anterior pituitary galanin- and neuromedin U-like immunoreactivity by use of immunocytochemistry and electron microscopy in rats in the normal state and after estrogen administration or adrenalectomy. In normal rats galanin immunoreactivity was present in a few corticotropes and lactotropes, females showing more than males; neuromedin U-like immunoreactivity was present in some thyrotropes and most corticotropes, in both sexes. Where galanin, neuromedin U and ACTH immunoreactivities were colocalized in corticotropes they were present in the same granules. Estrogen administration caused an increase in number of galanin immunoreactive lactotropes, as previously shown. The proportion of neuromedin U-positive corticotropes was not affected. After adrenalectomy, only females showed a significant increase in the proportion of galanin-positive corticotropes. Neuromedin U immunoreactivity was significantly increased in both sexes, as previously shown. Thus, in rat, as in man, galanin can be present in corticotropes and its expression appears to be sexrelated. This finding, and the demonstration of thyrotrope neuromedin U (only examined in normal females), provide correlation with previous experiments. The influence of endocrine status on the expression of these novel peptides underlines the inherent plasticity of pituitary endocrine cells.
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  • 71
    ISSN: 1432-0878
    Keywords: Proventriculus ; Endocrine secretory cells ; Secretory granules ; Peptide hormones ; Colocalization ; Immunocytochemistry ; Colloidal gold ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The endocrine cells of the chicken proventriculus were investigated immunocytochemically, using the peroxidase-antiperoxidase technique on paraffin and semithin sections for light microscopy, and immunogold staining in osmium-fixed material for electron microscopy. The fixation procedure also allowed a detailed ultrastructural investigation. Twenty-three antisera were tested and 7 immunoreactive cell-types were identified: D-cells containing somatostatin-like peptide; EG-cells immunoreactive to anti-glucagon, anti-GLP1 and antineurotensin; NT-cells labelled only with anti-neurotensin; BN-cells containing bombesin-like material; ENK-cells showing met-enkephalin immunoreactivity; EC-cells reactive to anti-serotonin; and APP-cells positive to anti-avian pancreatic polypeptide. In addition, enterochromaffin-like (ECL) cells, were also detected by electron microscopy. The presence of ENK-cells and the ultrastructure of these and NT-cells are described for the first time in chicken proventriculus, and glucagon, GLP1 and neurotensin are shown to be colocalized in the EG-cells.
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  • 72
    ISSN: 1432-0878
    Keywords: Neuroendocrine cells ; Intestine ; Chromogranin ; Serotonin ; Regulatory peptides ; Immunocytochemistry ; Podarcis hispanica (Reptilia, Lacertilia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Numerous endocrine cells can be observed in the gut of the lizard Podarcis hispanica after application of the Grimelius silver nitrate technique. The argyrophilic endocrine cells are usually tall and thin in the small intestine but short, basal, and round in the large intestine. Eleven types of immunoreactive endocrine cells have been identified by immunocytochemical methods. Numerous serotonin-, caerulein/gastrin/cholecystokinin octapeptide-and peptide tyrosine-tyrosine-immunoreactive cells; a moderate number of pancreatic polypeptide-, neurotensin-, somatostatin-, glucagon-like peptide-1-and glucagon-immunoreactive cells, and few cholecystokinin N-terminal-and bombesin-immunoreactive cells were found in the epithelium of the small intestine. Coexistence of glucagon with GLP-1 or PP/PYY has been observed in some cells. In the large intestine a small number of serotonin-, peptide tyrosine-tyrosine-, pancreatic polypeptide-, neurotensin-, somatostatin-and glucagon-like peptide-1-immunoreactive cells were detected. Vasoactive intestinal peptide immunoreactivity was found in nerve fibers of the muscular layer. Substance P-immunoreactive nerve fibers were detected in lamina propria, submucosa and muscular layer. Chromogranin A-immunoreactive cells were observed throughout the intestine, although in lower numbers than argyrophilic cells.
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  • 73
    ISSN: 1432-0878
    Keywords: Stellate ganglion ; Calcitonin gene-related peptide (CGRP) ; Substance P ; Immunocytochemistry ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The colocalization of immunoreactivities to substance P and calcitonin gene-related peptide (CGRP) in nervous structures and their correlation with other peptidergic structures were studied in the stellate ganglion of the guinea pig by the application of double-labelling immunofluorescence. Three types of fibre were distinguished. (1) Substance P+/CGRP+ fibres, which sometimes displayed additional immunoreactivity for enkephalin, constituted a small fibre population of sensory origin, as deduced from retrograde labelling of substance P+/CGRP+ dorsal root ganglion cells. (2) Substance P+/CGRP− fibres were more frequent; some formed baskets around non-catecholaminergic perikarya that were immunoreactive to vasoactive intestinal polypeptide (VIP). (3) CGRP+/substance P− fibres were most frequent and were mainly distributed among tyrosine hydroxylase (TH)-immunoreactive cell bodies. The peptide content of fibre populations (2) and (3) did not correspond to that of sensory ganglion cells retrogradely labelled by tracer injection into the stellate ganglion. Therefore, these fibres are throught to arise from retrogradely labelled preganglionic sympathetic neurons of the spinal cord, in which transmitter levels may have been too low for immunohistochemical detection of substance P or CGRP. CGRP-immunoreactivity but no substance P-immunolabelling was observed in VIP-immunoreactive postganglionic neurons. Such cell bodies were TH-negative and were spared by substance P-immunolabelled fibre baskets. Retrograde tracing with Fast Blue indicated that the sweat glands in the glabrous skin of the forepaw were the targets of these neurons. The streptavidin-biotin-peroxidase method at the electron-microscope level demonstrated that immunoreactivity to substance P and CGRP was present in dense-cored vesicles of 50–130 nm diameter in varicosities of non-myelinated nerve fibres in the stellate ganglion. No statistically significant difference in size was observed between vesicles immunolabelled for substance P and CGRP. Immunoreactive varicosities formed axodendritic and axosomatic synaptic contacts, and unspecialized appositions to non-reactive neuronal dendrites, somata, and axon terminals. Many varicosities were partly exposed to the interstitial space. The findings provide evidence for different pathways utilizing substance P and/or CGRP in the guinea-pig stellate ganglion.
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  • 74
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    Cell & tissue research 273 (1993), S. 79-87 
    ISSN: 1432-0878
    Keywords: Thymus ; Epithelial cells ; Oxytocin ; Immunocytochemistry ; Paracrine secretion ; Mouse (NMRI)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunoreactive oxytocin is expressed by thymic epithelial cells, which share properties with neuroendocrine cells. In order to investigate the assumed paracrine secretion of oxytocin, we studied the subcellular localization of immunoreactive oxytocin within thymic tissue and cultured thymic epithelial cells of the male mouse. Three types of immunoreactive cells were distinguished with the electron microscope. Immunoreactive oxytocin was found to be restricted to the cytoplasm by the use of pre- and postembedding methods. Some epithelial cells, especially in the cortex, showed a pronounced labelling of vesicular membranes and membrane tubules of the endoplasmic reticulum. In some cells, keratin filaments were associated with the electrondense stain. Under culture conditions immunoreactive cells of different shapes were found, all displaying similar patterns of labelling. The contents of different types of vacuoles were only rarely labelled. A special class of immunoreactive exocytotic vesicles could not be identified. Thus, our results do not support neuroendocrine secretion of oxytocin via vesicles of thymic epithelial cells but offer alternative modes of secretion.
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  • 75
    ISSN: 1432-0878
    Keywords: Lung ; Neuroepithelial bodies ; Immunocytochemistry ; 7B2 Protein ; Serotonin ; Rana temporaria (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The neuroendocrine cell population of the lung of Rana temporaria has been studied by means of immunocytochemistry. Serotonin (5HT)- and polypeptide 7B2-immunoreactive neuroepithelial bodies have been observed in the epithelial lining of the lung. 5HT- but not 7B2-immunoreactive isolated endocrine cells have also been observed.
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  • 76
    ISSN: 1432-0878
    Keywords: Blood group antigens ; Immunocytochemistry ; Cochlea ; Hair cells ; Development, ontogenetic ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The possible structure of human blood-group antigens, as found in cochlear hair cells of 3-day-old rats, is suggested. Data were obtained from immunocytochemical studies using 77 antibodies against the major human blood group antigens of the ABO, H, I and Lewis genetic systems. Neither the anti-A-related nor the anti-Lewis-related antibodies showed any positive immunoreaction on hair cells. In contrast, anti-B, anti-AB and anti-H antibodies displayed specific positive immunoreactive patterns on the hair cells. The results suggest that, in immature hair cells, two main glycoconjugate structures of the lactoseries are present: H type 2 antigen, which is the precursor of the B type 2 antigen, and the B type 2 antigen itself. Similar H and B carbohydrate structures have been reported in rat olfactory receptors. The type 2 glycoconjugates carrying these H and B antigens of auditive and olfactory receptors are resistant to fixation and paraffin embedding, suggesting that they might be glycoproteins. These auditive and olfactory H and B antigens must be different from the B-related antigens that are expressed by pseudo-unipolar neurons of rat posterior root ganglia, that are built from type 4 core chains, and that are destroyed by routine paraffin embedding procedures.
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  • 77
    ISSN: 1432-0878
    Keywords: Endothelin ; Nasal mucosa ; Immunocytochemistry ; Laser Doppler flowmetry ; Human ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Nasal blood flow is finely regulated by local release of neurotransmitters, neuropeptides and other bioactive molecules acting via paracrine mechanisms. We have investigated the occurrence and distribution in human nasal mucosa of endothelin, a potent vasoconstrictor peptide, by immunocytochemistry and the effect of systemic administration of endothelin-1 on vascular perfusion of rabbit nasal mucosa by laser Doppler flowmetry. Endothelin-like immunoreactivity was demonstrated within vascular endothelial cells in both developing and mature human mucosa. Nasal epithelial cells and some connective tissue cells, presumed to be macrophages, also displayed specific immunostaining. In rabbits injected with endothelin-1, a potent and prolonged nasal vasoconstriction was observed. It is suggested that endothelin released locally may participate in the regulation of nasal blood flow via paracrine mechanisms. Since endothelin has growth-promoting actions on several cell types, it is also tentatively proposed that this regulatory peptide may play a role during development of the nose.
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  • 78
    ISSN: 1432-0878
    Keywords: Intraepithelial axons ; Free nerve endings ; Nasal mucosa ; Ultrastructure ; Enzyme histochemistry ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of nerve fibres in the mucosa of the nasal septum of the rat was investigated by means of transmission electron microscopy on transverse and tangential ultrathin sections. Near the basement membrane of respiratory and squamous epithelium, a rather dense network of unmyelinated nerve fibres occurs. Some fibres in the respiratory epithelium ascend between the epithelial cells to reach up to the tight junctions. These fibres appeared in transverse sections to end as hooks or boutons, sometimes with branches. These shapes resemble the free nerve endings that are considered to act as nociceptors. The small intraepithelial fibres, with diameters of about 0.5–1 μm, contain both dense granules and clear vesicles comparable to synaptic vesicles. Substance P was found in dense granules in basal fibres; vasoactive intestinal peptide was absent throughout the epithelium. Acetylcholinesterase activity was observed closely associated with the basal fibres; the apical fibres showed little if any activity. Membrane specializations pointing to an efferent function as well as structures usually associated with mechanoreceptive functions were lacking in both respiratory and squamous epithelium.
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  • 79
    ISSN: 1432-0878
    Keywords: Gliobastoma multiforme ; Cell culture ; Glial fibrillary acidic protein ; Immunocytochemistry ; Cytogenetics ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Two human cell lines (GL15 and GL22) derived from glioblastoma multiforme were established and characterized by immunohistochemical and cytogenetic techniques. The expression of glial fibrillary acidic proteins and the karyotype were analyzed at different passages for both cell lines. The course of marker-pattern differed in the two cell lines. The main findings were a cell-density-dependent expression of glial fibrillary acidic protein in the cell line GL15 at all passages and a decreased expression of this protein over time in the cell line GL22. Both cell lines had hyperdiploid karyotypes and exhibited glioma-specific chromosomal abnormalities (gain of chromosome 7 and loss of chromosome 10). In the GL15 cell line no relevant chromosomal changes were produced during culturing, whereas in the GL22 cell line a hypodiploid clone appeared at the 42nd passage. The immunohistochemical and cytogenetic data resulting from this study confirm that the two cell lines established in our laboratory originated from astrocytic tumor cells.
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  • 80
    ISSN: 1432-0878
    Keywords: Pituitary adenylate cyclase-activating peptide (PACAP) ; Vasoactive intestinal peptide (VIP) ; Immunocytochemistry ; Respiratory tract ; Autonomic innervation ; Mammals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pituitary adenylate cyclase-activating peptide (PACAP) is a vasoactive intestinal peptide (VIP)-like peptide recently isolated from ovine hypothalami. Nerve fibers displaying PACAP immunoreactivity were found in the respiratory tract of rats, guinea pigs, ferrets, pigs, sheep and squirrel monkeys. A moderate supply of PACAP-immunoreactive fibers was seen in the nasal mucosa of guinea pigs. Few to moderate numbers of PACAP-containing fibers occurred in the tracheo-bronchial wall of rats, guinea pigs, ferrets, pigs, sheep and squirrel monkeys. The fibers were distributed beneath the epithelium, around blood vessels and seromucous glands, and among bundles of smooth muscle. In the lungs, the immunoreactive fibers were observed close to small bronchioli. A few PACAP-immunoreactive nerve cell bodies were seen in the sphenopalatine and otic ganglia of guinea pigs. Simultaneous double immunostaining of the respiratory tract of sheep and ferrets revealed that all PACAP-containing nerve fibers stored VIP. We suggest that neuronal PACAP may take part in the regulation of smooth muscle tone and glandular secretion.
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  • 81
    ISSN: 1432-0878
    Keywords: Endocrine cells, pulmonary ; Regulatory peptides ; Lung ; Immunocytochemistry ; Domestic fowl ; Domestic pigeon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract the presence of endocrine cells and nerves in the lung of 2 avian species (Gallus gallus and Columba livia domestica) has been studied by peroxidase-antiper-oxidase (PAP) and avidin-biotin complex (ABC) immunocytochemical methods at the light-microscopic level. Two immunoreactive cell-types have been identified in the epithelium of the primary and secondary bronchi of chick lung: serotonin- and bombesin-immunoreactive cells; and 3 cell-types, namely, serotonin-, bombesin- and CGRP-(calcitonin gene related peptide) immunoreactive cells, have been located in the bronchial epithelium of pigeon lung. Co-localization of 2 different immunoreactivities within the same cell has not been detected. VIP-immunoreactive nerves have been observed in different locations in chick lung.
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  • 82
    ISSN: 1432-0878
    Keywords: Brain, vertebrate ; Phenylethanolamine-N-methyltransferase ; Adrenaline ; Immunocytochemistry ; HPLC ; Rat (Wistar, Sprague-Dawley, Long Evans, Zucker)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemistry was used to compare the immunoreactivity of adrenergic neurons to a well characterized specific immunoserum to phenylethanolamine-N-methyltransferase (PNMT) in different strains of rats commonly used in research studies. In adult animals, marked differences were found in the PNMT-immunoreactivity of neurons between Wistar rats and other strains, resulting in a lower PNMT-immunostaining intensity (i) within neuronal perikarya of the medulla oblongata, and (ii) more strikingly, within nerve fibers and terminals located in various brain regions. This low PNMT-immunoreactivity of nerve fibers was detected both in 14- and 35-day-old Wistar rats. On the other hand, the HPLC measurement of catecholamines, in particular of adrenaline in the hypothalamus and the medulla oblongata, did not show any difference between adult Wistar and Sprague-Dawley rats. These data suggest that the low PNMT-immunoreactivity observed in central adrenergic neurons of the Wistar rats is related to the poor recognition of the antigen by the PNMT-antibody used. Possibly, these nerve cells mainly display an isoform of the enzyme that is immunologically different from the PNMT contained within the adrenergic neurons of other rat strains.
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  • 83
    ISSN: 1432-0878
    Keywords: Cardioactive peptide ; Immunocytochemistry ; Bivalve ; Gastropod ; Pulmonate ; Opisthobranch ; Lymnaea stagnalis (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The VD1 and RPD2 neurons of Lymnaea stagnalis innervate other central neurons, certain skin areas, the pneumostome area, and the auricle of the heart. Recently, a set of four (λ, ω, α, β) neuropeptides produced by these giant neurons and by certain other central neurons has been characterized. Although alternative splicing of the preprohormone of these neurons yields at least 10 different α neuropeptides, an affinity-purified antiserum directed against a domain common to all α neuropeptides has previously been shown to be highly selective in staining VD1, RPD2 and other neurons that produce the preprohormone. Since the gene encoding the neuropeptides is structurally similar to that expressed in R15 of the marine opisthobranch Aplysia californica, we have used the affinity purified antiserum as a marker for VD1/RPD2-related systems in other molluscs. Immunopositive neurons and fibers are observed in the central nervous systems of all species studied (Achatina fulica, Anodonta sp., Aplysia brasiliana, A. californica, Bulinus truncatus, Cepea sp., Eobania vermiculata, Helix aspersa, H. pomatia, Limax maximus, Mytilus edulis, Nassarius reticulatus, Viviparus viviparus). Several medium-sized and small neurons and 1–4 giant neurons are found in the pulmonates and opisthobranchs. The giant neurons in pulmonates have locations in the subesophageal ganglion, axonal branching patterns, and terminal arborizations in the auricle of the heart; all these characteristics are similar to those of VD1 and RPD2. Double-labelling (Lucifer yellow injection, immunocytochemistry) confirms that the two giant neurons in Helix pomatia are Br and Br. The immunoreactive cells in A. fulica appear to include the VIN and PON neurons. The antiserum also stains cells that appear to be the R15 neurons in two Aplysia species. The small and medium-sized neurons are distributed widely over the central ganglia of opisthobranchs and pulmonates. In prosobranchs and bivalves, small neurons are found in the cerebral and abdominal ganglia. No evidence has been found for innervation of the heart in these latter groups but in M. edulis, immunoreactive terminals can be observed in the gill. The results suggest the evolutionary conservation of immunoreactive peptides and the neurons that produce them, and thus support and extend previous hypotheses regarding the homology of certain giant neurons across molluscan species.
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  • 84
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Rathke's pouch ; Intermediate filaments ; Cytokeratins ; Development, ontogenetic ; Immunocytochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An immunohistochemical study of the production of the intermediate filaments [vimentin, cytokeratin, and glial filament acidic protein (GFAP)] during development of the pituitary gland was made by use of fetal and adult human pituitary tissue. Among these intermediate filament proteins in the anterior and intermediate lobes of the pituitary, cytokeratin is the first to appear, followed by GFAP and vimentin. However, only cytokeratin is seen during the period of morphogenesis of the pituitary gland, with the type-II subfamily cytokeratin 8 being the earliest to appear. Among the simple-epithelial-type cytokeratins, cytokeratins 8 and 19 were observed within the pituitary primordium during morphogenesis. Cells immunoreactive for cytokeratins 8 and 19 showed a heterogeneous three-dimensional distribution pattern in Rathke's pouch. Both cytokeratins 8 and 19 tended to be strongly positive at sites in the pituitary primordium where cells had become more loosely arranged (i.e., areas far from the diencephalon) but were only weakly positive in areas in which the epithelial cells were densely packed (i.e., areas closely associated with the diencephalon). It is concluded that, during the period of morphogenesis, Rathke's pouch has the intermediate filaments characteristic of simple epithelium and shows different immunoreactivity for simple-epithelial-type cytokeratins from place to place according to the extent of cellular differentiation.
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  • 85
    ISSN: 1432-0878
    Keywords: Cereborospinal fluid-contacting neurons ; Vasoactive intestinal peptide (VIP) ; Immunocytochemistry ; Nucleus accumbens/lateral septum ; Photoreceptors, extraocular ; Rod-opsin ; Cone-opsin ; Clemmys leprosa (Chelonia) ; Varanus monitor, Lacerta sicula (Lacertilia) ; Python reticulatus (Serpentes) ; Crocodylus niloticus (Crocodilia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract By means of immunocytochemical demonstration of vasoactive intestinal peptide (VIP) an accumulation of cerebrospinal fluid (CSF)-contacting neurons was found in a circumscribed region of the nucleus accumbens/lateral septum of eleven reptilian (chelonian, lacertilian, ophidian, crocodilian) species. Basal processes of these cells contribute to a subependymal plexus whose density displays considerable interspecific variation. VIP-immunoreactive nerve fibers occur also in the lateral septum and the nucleus accumbens where they encompass immunonegative cells in a basket-like pattern. The CSF-contacting neurons are surrounded by columnar ependymocytes frequently arranged in a pseudostratified manner. These specialized arrays of ependymal cells, however, occupy a more extended area than the VIP-immunoreactive CSF-contacting neurons and can be traced from the rostro-ventral pole of the lateral ventricle to the interventricular foramen. These observations suggest the existence of a telencephalic site of CSF-contacting neurons which may be more widespread than hitherto thought and which may participate in a circumventricular system of the lateral ventricle. Previous studies mainly performed with birds indicate that the VIP-immunoreactive CSF-contacting neurons of the nucleus accumbens might form a part of the “encephalic” (extraretinal and extrapineal) photoreceptor. However, further experiments are required to test this supposition since the VIP-immunoreactive neurons of the nucleus accumbens remained unlabeled by antibodies against bovine rodopsin and chicken cone-opsin in all eleven species analysed in this investigation.
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  • 86
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    Cell & tissue research 274 (1993), S. 127-133 
    ISSN: 1432-0878
    Keywords: Spectrin ; Cytoskeleton ; Isoforms ; Heart ; Immunocytochemistry ; Western blotting ; Mouse (C57BL/6)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of two isoforms of spectrin in the adult mouse heart was investigated by Western blotting and immunocytochemistry by use of monospecific antibodies to erythrocyte spectrin and nonerythroid brain spectrin (240/235). Western blotting revealed proteins analogous to both isoforms of α-spectrin in adult heart. Light-microscopic immunocytochemistry indicated that erythroid spectrin was distributed throughout the myocardium, with immunofluorescence localized to plasma membranes, Z-lines, and intercalated discs. Antibodies to brain spectrin (240/235) exhibited staining throughout the heart, with a generally diffuse distribution except for the prominent immunoreactivity associated with the intercalated discs. Nonerythroid spectrin immunofluorescence was detected in the endothelial cells of the endocardium and the mesothelial cell lining of the epicardium. Erythrocyte spectrin was not detected in the endocardium or the epicardium. The identification and localization of spectrin isoforms in the mammalian heart suggest the importance of spectrin proteins in the structural integrity and proper function of cardiac cells and tissues. This is the first demonstration of two different α-spectrin subunits in the mammalian heart.
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  • 87
    ISSN: 1432-0878
    Keywords: Clotting factors ; Antimicrobial substances ; Hemocytes ; Immunocytochemistry ; Tachypleus tridentatus (Chelicerata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The structure of hemocytes in the normal state and during blood coagulation, and the intracellular localization of three clotting factors and two antimicrobial factors were examined in the Japanese horseshoe crabTachypleus tridentatus. Two types of hemocytes were found in the circulating blood: non-granular and granular hemocytes. The latter contained numerous dense granules classed into two major types: L- and D-granules. The L-granules were larger (up to 1.5 μm in diameter) and less electron-dense than the D-granules (less than 0.6 μm in diameter). The L-granules contained three clotting factors and one antimicrobial factor, whereas the D-granules exclusively contained the other antimicrobial factor. After treatment with endotoxin, the L-granules were released more rapidly than the D-granules, although almost all granules were finally exocytosed. The granular hemocyte possessed a single Golgi complex; possible precursor granules of L-granules and D-granules contained tubular and condensed dense material, respectively. These data are discussed in relation to the self-defense mechanisms of the horseshoe crab.
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  • 88
    ISSN: 1432-0878
    Keywords: Gonadotropin ; Gonadotropin subunits ; Gonadotropes ; In situ hybridization ; Immunocytochemistry ; Pituitary gland, pars distalis ; Oogenesis ; Oncorhynchus mykiss (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Biosynthesis of salmon gonadotropins, GTH I and GTH II, during ovarian development, were examined by means of in situ hybridization histochemistry and indirect immunocytochemistry. In rainbow trout pituitary glands, expression of GTH Iβ- and IIβ-subunit genes appeared separately in distinct cells (GTH I- and GTH II-cells), whereas the GTH α-subunit gene was expressed in both cell-types. In the GTH I-cells, coordinated increases in GTh, α and Iβ messenger ribonucleic acids (mRNAs) occurred coincident with the onset of vitellogenesis, indicating active synthesis of GTH I during vitellogenesis. In contrast, in the GTH II-cells, both GTH α-and IIβ-mRNA signals markedly increased from a later stage of vitellogenesis and persisted throughout oocyte maturation and ovulation, supporting the idea that GTH II is actively synthesized as a maturational GTH. GTH α-mRNA levels in the GTH I-cells selectively decreased prior to final oocyte maturation, although Iβ-mRNA levels remained elevated, thus suggesting a decline of biosynthesis of GTH I after vitellogenesis. These findings clarify how the synthesis of GTH I and GTH II are coordinated in the piscine pituitary, and indicate that the expression of GTH subunit genes during gametogenesis is regulated differentially in a cell-specific manner, both temporally and spatially.
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  • 89
    ISSN: 1432-0878
    Keywords: Visual system ; Neuropeptides ; Serotonin ; Pigment-dispersing hormone ; Immunocytochemistry ; Neuromodulators ; Phormia terraenovae (Insecta) ; Leucophaea maderae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The pigment-dispersing hormone (PDH) family of neuropeptides comprises a series of closely related octadecapeptides, isolated from different species of crustaceans and insects, which can be demonstrated immunocytochemically in neurons in the central nervous system and optic lobes of some representatives of these groups (Rao and Riehm 1989). In this investigation we have extended these immunocytochemical studies to include the blowfly Phormia terraenovae and the cockroach Leucophaea maderae. In the former species tissue extracts were also tested in a bioassay: extracts of blowfly brains exhibited PDH-like biological activity, causing melanophore pigment dispersion in destalked (eyestalkless) specimens of the fiddler crab Uca pugilator. Using standard immunocytochemical techniques, we could demonstrate a small number of pigment-dispersing hormone-immunoreactive (PDH-IR) neurons innervating optic lobe neuropil in the blowfly and the cockroach. In the blowfly the cell bodies of these neurons are located at the anterior base of the medulla. At least eight PDH-IR cell bodies of two size classes can be distinguished: 4 larger and 4 smaller. Branching immunoreactive fibers invade three layers in the medulla neuropil, and one stratum distal and one proximal to the lamina synaptic layer. A few fibers can also be seen invading the basal lobula and the lobula plate. The fibers distal to the lamina appear to be derived from two of the large PDH-IR cell bodies which also send processes into the medulla. These neurons share many features in their laminamedulla morphology with the serotonin immunoreactive neurons LBO-5HT described earlier (see Nässel 1988). It could be demonstrated by immunocytochemical double labeling that the serotonin and PDH immunoreactivities are located in two separate sets of neurons. In the cockroach optic lobe PDH-IR processes were found to invade the lamina synaptic region and form a diffuse distribution in the medulla. The numerous cell bodies of the lamina-medulla cells in the cockroach are located basal to the lamina in two clusters. Additional PDH-IR cell bodies could be found at the anterior base of the medulla. The distribution and morphology of serotonin-immunoreactive neurons in the cockroach lamina was found to be very similar to the PDH-IR ones. It is hence tempting to speculate that in both species the PDH-and serotonin-immunoreactive neurons are functionally coupled with common follower neurons. These neurons may be candidates for regulating large numbers of units in the visual system. In the flies photoreceptor properties may be regulated by action of the two set of neurons at sites peripheral to the lamina synaptic layer, possibly by paracrine release of messengers.
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  • 90
    ISSN: 1432-0878
    Keywords: Corticotropin-releasing factor (CRF) ; Immunocytochemistry ; Arginine vasotocin (AVT) ; Mesotocin (MST) ; Co-localization ; Natrix maura (Serpentes)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The anatomical distribution of neurons and nerve fibers containing corticotropin-releasing factor (CRF) has been studied in the brain of the snake, Natrix maura, by means of immunocytochemistry using an antiserum against rat CRF. To test the possible coexistence of CRF with the neurohypophysial peptides arginine vasotocin (AVT) and mesotocin (MST) adjacent sections were stained with antisera against the two latter peptides. CRF-immunoreactive (CRF-IR) neurons exist in the paraventricular nucleus (PVN). In some neurons of the PVN, coexistence of CRF with MST or of CRF with AVT has been shown. Numerous CRF-IR fibers run along the hypothalamo-hypophysial tract and end in the outer layer of the median eminence. In addition, some fibers reach the neural lobe of the hypophysis. CRF-IR perikarya have also been identified in the following locations: dorsal cortex, nucleus accumbens, amygdala, subfornical organ, lamina terminalis, nucleus of the paraventricular organ, nucleus of the oculomotor nerve, nucleus of the trigeminal nerve, and reticular formation. In addition to all these locations CRF-IR fibers were also observed in the lateral septum, supraoptic nucleus, habenula, lateral forebrain bundle, paraventricular organ, hypothalamic ventromedial nucleus, raphe and interpeduncular nuclei.
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  • 91
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    Cell & tissue research 273 (1993), S. 1-29 
    ISSN: 1432-0878
    Keywords: Neuropeptides ; Neurohormones ; Neuromodulators ; Insect brain ; Immunocytochemistry ; Drosophila melanogaster, Calliphora vomitoria, Leucophaea maderae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
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  • 92
    ISSN: 1432-0878
    Keywords: SALMFamide ; Neuropeptide ; Immunocytochemistry ; Mapping ; Asterias rubens (Echinodermata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The recent isolation and characterization of the SALMFamide neuropeptides S1 and S2 from the starfish Asterias rubens has initiated a series of studies on their distribution. Specific antisera have been raised against S1 and used in light-microscopical immunocytochemistry. The results of this study reveal for the first time a possible hyponeural innervation of the visceral musculature of the gut and the widespread neuronal distribution of S1, (i) in axons and cell bodies of both ectoneural and hyponeral regions of the radial nerve cord and circumoral nerve ring, (ii) in the nerve ring and nerve plexus of the tube feet, (iii) in the apical muscle, (iv) in skin, and (v) extensively throughout the digestive system. These discoveries are of particular interest in terms of the possible functional roles for S1 in Asterias rubens.
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  • 93
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Secretory pathway ; C1B8A8 monoclonal antibody ; Immunocytochemistry ; Immunoaffinity chromatography ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The secretory pathway of the complex-type glycoprotein specific to the subcommissural organ (SCO) was examined using the monoclonal antibody (Mab) C1B8A8. Immunoreactive material was revealed in various compartments of the secretory ependymocyte, i.e., the endoplasmic reticulum, the Golgi area and the secretory vacuoles. In addition, immunoreactive material was also observed in the ventricular cavity. Evidence of a release both at the apical lining and at the basal process of the SCO ependymocytes suggests that the same protein could be secreted into the cerebrospinal fluid and the perivascular spaces. After immunoaffinity chromatography of soluble extracts of the SCO on Mab C1B8A8 immunoadsorbent columns, three glycopeptides were identified on Western blots; they were concanavalin A (Con A)-positive (88, 54 and 34 kDa) and wheat-germ agglutinin (WGA)-positive (54 and 34 kDa). The Con A-positive glycopeptide (88 kDa) is probably related to the high-mannose-type glycoprotein, the precursor form of the secreted compound, whereas the 54 kDa-glycopeptide that is both Con A- and WGA-positive could represent an intermediate form. The 34 kDa-glycopeptide that is strongly WGA-positive could be related to the monomeric form of the secreted compound. These three glycopeptides were not revealed in eluted fractions of soluble extracts of the ependyma that served as control.
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  • 94
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    Cell & tissue research 271 (1993), S. 375-380 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Gut ; Innervation ; Regulatory peptides ; Endocrine cells ; Marthasterias glacialis (Echinodermata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The endocrine cells of the starfish digestive tract are spindle-shaped, contacting both the lumen and the basiepithelial plexus. Silver impregnation labels the basiepithelial and subcoelomic plexuses as well as these cells. Twenty antisera have been tested using the avidinbiotin method, in order to identify the regulatory substances involved in this system. Endocrine cells and nerves immunoreactive to GFNSALMFamide- (S1), FMRFamide-, peptide tyrosine-tyrosine-(PYY), pancreatic polypeptide- (PP), melanocyte stimulating hormone- (αMSH) and peptidylglycine alpha-amidating monooxygenase- (PAM) specific antisera have been found in the epithelium. The antibodies against S1, a peptide isolated from the nervous system of a starfish, and αMSH, stain both the basiepithelial plexus and the subcoelomic plexus, but the others react only with nerves in the basiepithelial plexus. Absorption controls show that antibodies for S1 and FMRFamide totally crossreact recognizing the same molecule, possibly S1. The other antibodies do not show cross-reactivity to any of the rest, and thus we conclude that these regulatory peptides are present in starfish. This is the first report of the presence of FMRFamide, PYY, αMSH and PAM in the Echinodermata. Under the electron microscope the endocrine cells exhibit secretory granules, microtubules and mitochondria. Direct contact with the subcoelomic plexus can be observed.
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  • 95
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    Cell & tissue research 263 (1991), S. 71-79 
    ISSN: 1432-0878
    Keywords: Aromatase ; Immunocytochemistry ; Brain aromatase ; Preoptic area ; Hypothalamus ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of aromatase-immunoreactive cells was studied by immunocytochemistry in the mouse forebrain using a purified polyclonal antibody raised against human placental aromatase. Labeled perikarya were found in the dorso-lateral parts of the medial and tuberal hypothalamus. Positive cells filled an area extending between the subincertal nucleus in the dorsal part, the ventromedial hypothalamic nucleus in the ventral part, and the internal capsule and the magnocellular nucleus of the lateral hypothalamus in the lateral part. The same distribution was seen in the two strains of mice that were studied (Jackson and Swiss), and the number of immunoreactive perikarya did not seem to be affected by castration or testosterone treatment. No immunoreactivity could be detected in the medial regions of the preoptic area and hypothalamus; these were expected to contain the enzyme based on assays of aromatase activity performed in rats and on indirect autoradiographic evidence in mice. Our data raise questions concerning the distribution of aromatase in the brain and the mode of action of the centrally produced estrogens.
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  • 96
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    Cell & tissue research 263 (1991), S. 195-198 
    ISSN: 1432-0878
    Keywords: Pancreas, endocrine ; Islets of Langerhans ; Immunocytochemistry ; Endocrine cells four types ; Electron microscopy ; Sminthopsis crassicaudata (Marsupialia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The endocrine pancreas of the Australian fattailed dunnart, Sminthopsis crassicaudata, was investigated by means of electron-microscopic immunocytochemistry using the protein A-gold technique on London resin (LR) white-embedded tissue. The primary antibodies used were raised against insulin, glucagon, somatostatin and pancreatic polypeptide. The morphology of the secretory granules differed in the four cell types. The insulin cells are pleomorphic, and the secretory granules composed of an electron-dense core surrounded by an electron-lucen halo. The glucago cells possess granules with an electron-dense core usually surrounded by a halo of less dense granular material. Somatostatin cells have large, less dense secretory granules. The pancreatic polypeptide cells show small, dense secretory granules. In order for an ultrastructural study to be considered reliable for the definite identification of endocrine cell types, it is essential that it be corroborated by immunocytochemical data at the light-or preferably electron-microscopic level. Recent developments in immuno-electron-microscopic techniques have contributed to a better knowledge of cells responsible for the secretion of a wide variety of hormones, as in this study.
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  • 97
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    Cell & tissue research 272 (1993), S. 183-192 
    ISSN: 1432-0878
    Keywords: Retina ; Müller cells ; Neuron-specific enolase ; Immunocytochemistry ; Quantitative analysis ; Ultrastructure ; Bufo marinus (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have previously shown that an antibody against neuron-specific enolase (NSE) selectively labels Müller cells (MCs) in the anuran retina (Wilhelm et al. 1992). In the present study the light- and electron-microscopic morphology of MCs and their distribution were described in the retina of the toad, Bufo marinus, using the above antibody. The somata of MCs were located in the proximal part of the inner nuclear layer and were interconnected with each other by their processes. The MCs were uniformly distributed across the retina with an average density of 1500 cells/mm2. Processes of MCs encircled the somata of photoreceptor cells isolating them from each other by glial sheath, except for those of the double cones. Some of the photoreceptor pedicles remained free of glial sheath. Electron-microscopic observations confirmed that MC processes provide an extensive scaffolding across the neural retina. At the outer border of the ganglion cell layer these processes formed a non-continuous sheath. The MC processes traversed through the ganglion cell layer and spread beneath it between the neuronal somata and the underlying optic axons. These processes formed a continuous inner limiting membrane separating the optic fibre layer from the vitreous tissue. Neither astrocytic nor oligodendrocytic elements were found in the optic fibre layer. The significance of the uniform MC distribution and the functional implications of the observed pattern of MC scaffolding are discussed.
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  • 98
    ISSN: 1432-0878
    Keywords: Pineal organ ; Epithalamus ; Pinealocytes ; Hydroxyindole O-methyltransferase (HIOMT) ; Immunocytochemistry ; HIOMT-mRNA ; In-situ hybridization ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Hydroxyindole O-methyltransferase (HIOMT)-immunoreactive cells and melatonin synthesis were demonstrated in bovine epithalamus (including the pineal gland) with monoclonal antibodies and cRNA probes to HIOMT. The HIOMT-immunoreactive product was present in the cytoplasm of pinealocytes. All identifiable pinealocytes were clearly labeled in the pineal gland. The expression of the HIOMT gene was first identified in pinealocyte cytoplasm by in-situ hybridization (ISH). The distribution of the hybridization-positive cells in the pineal gland was compatible with that revealed by immunocytochemistry using the monoclonal antibody to HIOMT. In addition, HIOMT transcripts were found in the medial habenular nucleus, and the habenular and posterior commissure; they may correspond to S-antigen-immunoreactive cells demonstrated in the same regions of the hamster and the mouse. In these regions, the hybridization-positive cells did not exhibit HIOMT-immunoreactivity; thus, cells devoid of immunoreactivity may synthesize but rapidly transport the newly synthesized proteins. These results indicate (1) that the conversion of N-acetylserotonin into melatonin takes place in the cytoplasm of pinealocytes, (2) that some epithalamic cells in the habenular area may synthesize melatonin, and (3) that melatonin may act as a chemical messenger in centrally directed processes, as shown by using S-antigen immunocytochemistry.
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  • 99
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    Cell & tissue research 272 (1993), S. 383-389 
    ISSN: 1432-0878
    Keywords: Mammary gland ; Growth inhibitor ; Epithelium ; Cell types ; Differentiation ; Immunocytochemistry ; Immunohistochemistry ; Cow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Localization of a mammary-derived growth inhibitor (MDGI) in the bovine mammary gland was verified by light-and electron-microscopic methods. Expression of MDGI, which is known to inhibit the growth of mammary epithelial cell lines in vitro, was found to be highest in the late pregnant and in the lactating state. A combination of immunohistochemical and immunocytochemical methods with semi- and ultrathin resin sections revealed marked variations in MDGI staining. High MDGI levels were predominantly detectable in epithelial cells with large milk fat droplets. Distinct cell types that were almost free of label could be identified among bovine mammary epithelial cells that always exhibited high MDGI levels. Similar results were obtained when using a serum-free organ culture system in which MDGI was hormonally induced in cell types of comparable differentiation state. The specific occurrence of the growth inhibitor in developing alveoli and certain cell types points to the association between MDGI expression and functional differentiation in the normal mammary gland.
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  • 100
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Cell-types ; Hypothalamoneurohypophysial system ; Immunocytochemistry ; Oncorhynchus keta (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The ontogeny of pituitary cell-types, hypothalamic neurons and their fibers, was studied immunocytochemically during development of chum salmon. Five weeks after fertilization (eyed stage embryos), 5 cell-types were detected in the adenohypophysial (AHP) anlage; prolactin (PRL)-, growth hormone (GH)-, adrenocorticotropin (ACTH)-, melanotropin (α-MSH)-, and thyrotropin (TSH)-producing cells. The PRL-, GH- and ACTH-cells were relatively well developed as compared with MSH- and TSH-cells. Gonadotropes, however, were not detected even 3 weeks after hatching (10 weeks after fertilization). The neurohypophysis (NHP), on the other hand, began to grow around the eyed stage. Neuroendocrine fibers, not only from the tuberal hypothalmus (melanin-concentrating hormone) but also from the preoptic regions (vasotocin and somatostatin), reached the NHP during the last week of embryonic life. In the developing pituitary, the ratio between the length of the boundary between the AHP and the NHP, and the area of the AHP, was stable, being approximately 1–4%. The coordinated development of the AHP and NHP in chum salmon seems to result in the development of the characteristic hypothalamo-hypophysial relationship by the time of hatching.
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