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  • Immunocytochemistry  (46)
  • Springer  (46)
  • American Institute of Physics
  • Annual Reviews
  • 2005-2009
  • 1990-1994  (46)
  • 1980-1984
  • 1945-1949
  • 1993  (46)
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  • 2005-2009
  • 1990-1994  (46)
  • 1980-1984
  • 1945-1949
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  • 1
    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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  • 2
    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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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    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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  • 4
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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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  • 5
    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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  • 6
    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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  • 7
    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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  • 8
    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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  • 9
    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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  • 10
    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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  • 11
    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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  • 12
    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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  • 13
    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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  • 14
    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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  • 15
    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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  • 16
    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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  • 17
    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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  • 18
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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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  • 19
    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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  • 20
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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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  • 21
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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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  • 22
    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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  • 23
    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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  • 24
    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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  • 25
    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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  • 26
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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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  • 27
    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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  • 28
    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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  • 29
    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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  • 30
    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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  • 31
    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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  • 32
    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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  • 33
    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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  • 34
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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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  • 35
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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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  • 36
    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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  • 37
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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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  • 38
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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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  • 39
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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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  • 40
    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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  • 41
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    Cell & tissue research 272 (1993), S. 459-471 
    ISSN: 1432-0878
    Keywords: Oocyte ; Yolk ; Receptor ; Endocytosis ; Tracer studies ; Immunocytochemistry ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract During the rapid final stage of growth, chicken oocytes take up massive amounts of plasma components and convert them to yolk. The oocyte expresses a receptor that binds both major yolk lipoprotein precursors, vitellogenin (VTG) and very low density lipoprotein (VLDL). In the present study, in vivo transport tracing methodology, isolation of coated vesicles, ligand- and immuno-blotting, and ultrastructural immunocytochemistry were used for the analysis of receptor-mediated yolk formation. The VTG/VLDL receptor was identified in coated profiles in the oocyte periphery, in isolated coated vesicles, and within vesicular compartments both outside and inside membrane-bounded yolk storage organelles (yolk spheres). VLDL particles colocalized with the receptor, as demonstrated by ultrastructural visualization of VLDL-gold following intravenous administration, as well as by immunocytochemical analysis with antibodies to VLDL. Lipoprotein particles were shown to reach the oocyte surface by passage across the basement membrane, which possibly plays an active and selective role in yolk precursor accessibility to the oocyte surface, and through gaps between the follicular granulosa cells. Following delivery of ligands from the plasma membrane into yolk spheres, proteolytic processing of VTG and VLDL by cathepsin D appears to correlate with segregation of receptors and ligands which enter disparate sub-compartments within the yolk spheres. In small, quiescent oocytes, the VTG/VLDL receptor was localized to the central portion of the cell. At onset of the rapid growth phase, it appears that this pre-existing pool of receptors redistributes to the peripheral region, thereby initiating yolk formation. Such a redistribution mechanism would obliterate the need for de novo synthesis of receptors when the oocyte's energy expenditure is to be utilized for plasma membrane synthesis, establishment and maintenance of intracellular topography and yolk formation, and preparation for ovulation.
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  • 42
    ISSN: 1432-0878
    Keywords: Pituitary gland, pars distalis ; Pars tuberalis ; Somatotrope heterogeneity ; Immunocytochemistry ; European ferret, Mustela putorius furo (Carnivora)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Light-microscopic immunocytochemistry of ferret anterior pituitary revealed the localization of somatotropes in the pars distalis, but no immunoreactive cells were detected in the pars tuberalis. Ultrastructural studies by superimposition immunocytochemistry and immuno-electron microscopy, clucidated the morphological heterogeneity of these somatotropic cells. They were classified into 2 subtypes on the basis of size of the secretory granules. Type-I cells with small granules (mean diameter, 192 nm), were considered to be the immature somatotrop, while Type-II cells, with comparatively larger secretory granules (mean diameter, 257 nm), were considered to be the matured form of Type-I cells and the typical somatotropic cell-type, and were much more predominant than the Type-I cells. The fact that Type-II cells had a distinct Golgi zone and many mitochondria, while in Type-I cells the intracellular organelles were generally less developed, supports this suggestion. In addition to these two extreme subtypes, several intermediate forms were also encountered that may represent different transitional phases during the conversion of Type I to Type II. Protein A-gold immuno-electron microscopy illustrated the specific localization of growth hormone over the granules, with no labelling over any other cytoplasmic organelles of the 2 somatotrope subtypes.
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  • 43
    ISSN: 1615-6102
    Keywords: Abscission ; Actin filaments ; Cytokinesis ; Immunocytochemistry ; Microtubules ; Moss protonema ; Preprophase band
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary When grown on nutrient agar, protonemata ofBryum tenuisetum produce aerial filaments containing several abscission or tmema cells (TC). Basipetal migration of the nucleus and some of the chloroplasts signals the onset of TC formation. This is followed by the creation of a plastid-free zone at the base of the mother cell. The ensuing cytokinesis produces a very short aplastidic TC. This expands without the deposition of new wall material. Eventually the wall ruptures around the equator thus disrupting the protonemal filament. The site of wall breakdown is marked by a narrow band of cortical cytoplasm containing colocalized circumferential rings of actin filaments and microtubules. A transverse band of microtubules appears at the extreme basal end of the tmema mother cell. This band, which is not colocalized with actin filaments, migrates distally over the surface of the nucleus. Intimate spatial and developmental correlations suggest that this transverse array of the microtubules has a key role in excluding plastids from the TC. It is therefore considered not to be homologous with a preprophase band.
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  • 44
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 176 (1993), S. 100-105 
    ISSN: 1615-6102
    Keywords: Bromodeoxyuridine ; Immunocytochemistry ; DNA synthesis ; Cell cycle ; In vitro culture ; Gigaspora margarita
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This study provides evidence thatGigaspora margarita replicates its nuclear DNA, even in the absence of a host plant. Three experimental approaches were used: (i) static cytofluorimetry to quantify the DNA content, (ii) pulse treatments with bromodeoxyuridine (BrdU), which is an analogue of thymidine, to reveal nuclei undergoing DNA synthesis, and (iii) ultrastructural observations to study changes in chromatin morphology during the fungal cell cycle. A slight second peak of approximately twice the value of a major peak was found by cytofluorimetry, showing that a small number of nuclei had entered in cycle during in vitro development. Nuclei which had incorporated BrdU were observed after pulses of 24 h; nuclei with condensed chromatin were also apparent at this time. The results demonstrate thatG. margarita has all the metabolic pathways needed to replicate its nuclear DNA even in the absence of the host, suggesting that more complex mechanisms inhibit the extended growth in vitro of arbuscular mycorrhizal fungi.
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  • 45
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 176 (1993), S. 1-13 
    ISSN: 1615-6102
    Keywords: Gloeomonas ; Immunocytochemistry ; Golgi apparatus ; Cell wall
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An antibody to the inner wall layer ofGloeomonas kupfferi was isolated and used in a developmental analysis of cell wall processing, secretion and extracellular assembly. The focus of the processing of this matrix layer is the endomembrane system, in particular the Golgi apparatus (GA) and contractile vacuole (CV). During interphase, inner wall materials are processed in the GA, packaged in trans face vesicles and transported to the CV, the final internal depository of wall precursors until release to the cell surface. During cell division, significant changes occur in the inner wall layer processing. Early on in cytokinesis, the GA does not label with our antibody, suggesting that other wall layers are being processed. In later stages of cytokinesis, the GA changes in morphology and begins to produce inner wall layer materials. These wall precursors are shuttled to the CV where they are released around the daughter cell protoplasts. The first wall layer that is formed around daughter cells is the crystalline median wall layer. Once assembled, the inner wall layer condenses upon the crystalline layer and grows in size.
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  • 46
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    Springer
    Environmental biology of fishes 38 (1993), S. 209-218 
    ISSN: 1573-5133
    Keywords: Terminal nerve ; Midbrain ; Immunocytochemistry ; Reproduction ; Brain ; Sharks ; Rays ; Skates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Synopsis Immunoreactive (ir) gonadotropin-releasing hormone (GnRH) is localized in many neurons of the terminal nerve (TN) and midbrain tegmentum, while few ir-cells are observed in the preoptic area and ventral hypothalamus. The paucity of preoptic ir-cells may relate to an unusual feature of the elasmobranch pituitary, i.e. a lack of portal control of gonadotropin-producing cells. TN and midbrain GnRH-ir neurons may be major sources of GnRH used to modulate or otherwise control both pituitary and brain cells via delivery through the systemic circulation. These ir-nuclei also appear to directly innervate CNS regions (the preoptic area, habenula and clasper control area of the spinal cord) involved in sexual functions. Important regulatory mechanisms, represented by interactions between GnRH pathways and sex-steroid concentrating neurons, are likely to occur in the preoptic area, habenula and midbrain tegmentum.
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