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  • Immunocytochemistry  (46)
  • Springer  (46)
  • American Chemical Society
  • Essen : Verl. Glückauf
  • Krefeld : Geologischer Dienst Nordhein-Westfalen
  • 2015-2019
  • 2005-2009
  • 1985-1989  (46)
  • 1980-1984
  • 2006
  • 1989  (46)
Collection
Publisher
  • Springer  (46)
  • American Chemical Society
  • Essen : Verl. Glückauf
  • Krefeld : Geologischer Dienst Nordhein-Westfalen
  • Wiley-Blackwell  (1)
Years
  • 2015-2019
  • 2005-2009
  • 1985-1989  (46)
  • 1980-1984
Year
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 258 (1989), S. 119-124 
    ISSN: 1432-0878
    Keywords: Development, ontogenetic ; Immunocytochemistry ; Monoclonal antibodies ; Mucosa ; Lymphoid organs ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The postnatal development of chicken mucosa-associated lymphoid tissues of the eyes, lungs, and intestines were investigated with monoclonal antibodies specific for either all leucocytes, B lymphocytes, mononuclear phagocytes, IgM, IgG, or IgA. Attention has been paid to the relation of lymphoid infiltrates with their surrounding mucosae, the segregation into B-cell and T-cell areas, development of germinal centers, and secretory immunoglobulins. Abudant secretory IgM and IgA was detected in the epithelium of the Harderian glands in the orbits, even though they lacked large leucocyte infiltrates with germinal centers. Lymphoid tissues in the mucosae of lungs and intestines developed separate B-cell and T-cell areas. The proventriculus, Meckel's diverticulum, and Peyer's patches generally contained germinal centers from 12 weeks of age on. Because chickens as young as 2 weeks old had germinal centers in bronchus-associated lymphoid tissue and cecal tonsils, these areas were probably highly stimulated by antigens. Isotype-specific monoclonal antibodies were used to detect IgM-, IgG-, and IgA-bearing follicular cells in the same germinal center.
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  • 2
    ISSN: 1432-0878
    Keywords: Insect AKH/RPCH ; Neurohormones ; Cam-HrTH-II ; Lom-AKH-I ; Immunocytochemistry ; Carausius morosus, Sarcophaga bullata (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A polyclonal antiserum was prepared against an N-terminal modified Cam-HrTH-II (Leu-Asn-Phe-...), one of the members of the large AKH/RPCH peptide family, first isolated from Carausius morosus. The localisation of this peptide was performed by means of immunocytochemical methods in the brain and corpora cardiaca-corpora allata complex of the stick insect, Carausius morosus and the grey fleshfly, Sarcophaga bullata. The distribution patterns of molecules reactive to the Cam-HrTH-II and the LomAKH-I antisera in both insect species were compared. In Carausius, both antisera reacted in the same cell bodies. In Sarcophaga, some neurons were stained by both, others only by one of the two antisera. By combining two different antisera, we demonstrated that there are no Lom-AKH-I-like molecules present in Carausius and that there must occur at least three different AKH-like molecules in the brain of Sarcophaga. One is similar to Cam-HrTH-II, the second to Lom-AKH-I and the third is an AKH/RPCH-like peptide, different from Lom-AKH-I and Cam-HrTH-II.
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  • 3
    ISSN: 1432-0878
    Keywords: Serotonin ; Immunocytochemistry ; Insect nervous system ; Protocerebrum ; Suboesophageal ganglion ; Manduca sexta (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Serotonin-immunoreactive neurons in the median protocerebrum and suboesophageal ganglion of the sphinx moth Manduca sexta were individually reconstructed. Serotonin immunoreactivity was detected in 19–20 bilaterally symmetrical pairs of interneurons in the midbrain and 10 pairs in the suboesophageal ganglion. These neurons were also immunoreactive with antisera against DOPA decarboxylase. All major neuropil regions except the protocerebral bridge are innervated by these neurons. In addition, efferent cells are serotonin-immunoreactive in the frontal ganglion (5 neurons) and the suboesophageal ganglion (2 pairs of neurons). The latter cells probably give rise to an extensive network of immunoreactive terminals on the surface of the suboesophageal ganglion and suboesophageal nerves. Most of the serotonin-immunoreactive neurons show a gradient in the intensity of immunoreactive staining, suggesting low levels of serotonin in cell bodies and dendritic arbors and highest concentrations in axonal terminals. Serotonin-immunoreactive cells often occur in pairs with similar morphological features. With one exception, all serotonin-immunoreactive neurons have bilateral projections with at least some arborizations in identical neuropil areas in both hemispheres. The morphology of several neurons suggests that they are part of neuronal feedback circuits. The similarity in the arborization patterns of serotonin-immunoreactive neurons raises the possibility that their outgrowing neurites experienced similar forces during embryonic development. The morphological similarities further suggest that serotonin-immunoreactive interneurons in the midbrain and suboesophageal ganglion share physiological characteristics.
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  • 4
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    Cell & tissue research 258 (1989), S. 101-109 
    ISSN: 1432-0878
    Keywords: Serotonin ; Immunocytochemistry ; Subesophageal ganglion ; Neurohemal organs ; Manduca sexta (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Serotonin-immunoreactivity is mapped in wholemounts and slices of the suboesophageal ganglion (SOG) of larval Manduca sexta by means of immunocytochemistry. An extensive meshwork of serotonin-immunoreactive nerve fibres on some peripheral nerves of the SOG has been demonstrated. This meshwork appears to belong to a serotonergic neurohemal system, probably supplied by two pairs of bilateral serotonin-immunoreactive neurons with big cell bodies on the dorsal side near the midline in the mandibular neuromere. Intracellular recording and staining revealed their physiology and morphology. These neurons produce long lasting (50 msec) action potentials, which suggest that they are neurosecretory cells. Two pairs of bilateral serotonin-immunoreactive interneurons similar to those of other insects are stained in the labial and maxillar neuromeres, but not in the mandibular neuromere. Their ventrolaterally located cell bodies project through a ventral commissure into the contralateral hemiganglion and then cross back again through a dorsal commissure. The axons project into the contralateral circumoesophageal connective.
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  • 5
    ISSN: 1432-0878
    Keywords: Na+, K+-ATPase ; Immunocytochemistry ; Kidney ; Salivary glands ; Transport ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antibody to the 96 kD α-subunit of the Na+, K+ -ATPase from Bufo marinus has been used in immunostaining rat kidney and salivary glands. Intense staining was observed on basolateral membranes of distal tubules of the kidney and striated ducts of the three major salivary glands. Less intense staining was seen on the basolateral membranes of parotid acinar cells, but no staining was seen on the acinar cells of submandibular or sublingual glands. These sites of staining have been shown, by other methods, to posses substantial Na+, K+ -ATPase, indicating that the antibody recognizes antigenic determinants of the sodium pump highly conserved in the course of evolution. In addition, staining with this antibody was observed at the apical region of cells of the proximal straight tubule and of the papillary collecting duct in the kidney. Absorption studies suggest that the apical antigenic determinants are the same or closely related to each other but are distinct from basolateral antigenic determinants.
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  • 6
    ISSN: 1432-0878
    Keywords: Met-enkephalin-Arg6-Phe7 ; Met-enkephalin-Arg6-Gly7-Leu8 ; Immunocytochemistry ; Neuropeptides ; Co-existence of peptides ; Neurosecretory cells, insects ; Blowfly, Calliphora vomitoria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neuronal pathways immunoreactive to antisera against the extended-enkephalins, Met-enkephalin-Arg6-Phe7 (Met-7) and Met-enkephalin-Arg6-Gly7-Leu8 (Met-8), have been identified in the brain of the blowfly Calliphora vomitoria. Co-localisation with other enkephalins in certain neurons suggests that a precursor similar to preproenkephalin A exists in insects and that differential enzymatic processing occurs as in vertebrates. Co-localisations of the extended-enkephalin-like peptides with other vertebrate-type peptides, including cholecystokinin and pancreatic polypeptide, also occur. The enkephalinergic pathways are specific, comprising a few groups of highly characteristic neurons and areas of neuropil. Of special interest is the finding that parts of the antennal chemosensory and the optic lobe visual systems contain Met-8 immunoreactive neurons. Within the median neurosecretory cell groups, some of the giant neurons show immunoreactivity to Met-8 and others to both Met-8 and Met-7. Fibres from these cells project to the corpus cardiacum and also to the suboesophageal ganglion, where arborisations occur in the tritocerebral neuropil. Co-localisation studies of these cells have shown that at certain terminals, one particular type of peptide is the dominant neuroregulator, whilst at other terminals, within the same cell, a different co-synthesised peptide predominates. Several groups of lateral neurosecretory cells show clearly defined enkephalinergic pathways, most of which have connections with the central body. The complex patterns of immunoreactivity seen in terminals in the different parts of the central body, suggest an important role for the enkephalin-like peptides in the integration of multimodal sensory inputs. The physiological functions of the extended-enkephalin-like peptides in the brain of Calliphora is still unknown, but the anatomical evidence suggests they may have a role similar to that in mammals, where they are thought to control aspects of feeding behaviour.
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  • 7
    ISSN: 1432-0878
    Keywords: Gut hormones ; Insulin ; Bombesin ; Immunocytochemistry ; Pancreatic polypeptide ; Cholecystokinin (CCK) ; Gastrin ; Schistocerca gregaria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The endocrine cells of the midgut epithelium of the desert locust are found dispersed among the digestive cells and are similar to those of the vertebrate gut. According to their reactivity to silver impregnation techniques and the ultrastructural features of the secretory granules (shape, electron-density, size, and structure) 10 types of endocrine cell have been identified, of which seven are located in the main segment of the midgut or in the enteric caeca, and the other three seem to be present only in the ampullae through which the Malpighian tubules drain into the gut. The endocrine cells have a slender cytoplasmic process that reaches the gut lumen, a feature that supports the receptosecretory nature postulated for this cellular type in insects as well as vertebrates. Antisera directed against mammalian gastrin, CCK, insulin, pancreatic polypeptide and bombesin reacted with some of the endocrine cells. This is the first time that insulin- and bombesin-like immunoreactive cells have been described in the midgut of an insect.
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  • 8
    ISSN: 1432-0878
    Keywords: Somatotropes, growth hormone cells ; Immunocytochemistry ; Growth hormone (GH) ; Receptors, membrane ; Somatostatin (SRIF) ; Growth hormone-releasing hormone (GRH) ; Rat (Han: WIST)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The synthetic peptides somatostatin (SRIF) and growth hormone-releasing hormone (GRH) were coupled directly to colloidal gold of different particle sizes. Both conjugates were biologically active in displacing the corresponding radiolabeled hormones from high affinity binding sites in pituitary membranes. Release of growth hormone (GH) from cultured anterior pituitary cells was modulated by both conjugates alone or in combination. Ultrastructural studies were performed with cells incubated at 4° C (2 h) and 37° C (2 min-2 h) with one of the labeled peptides or their combination. Somatotropes were identified by immunostaining with anti-rGH followed by protein A-ferritin, thus obtaining a triple labeling. Both hormone conjugates were internalized in different vesicles in the beginning but accumulated during longer incubation times in the same compartment. The secretory vesicles and the nucleus were not labeled by any hormone conjugate. In contrast to SRIF-gold, the uptake of GRH-gold conjugate decreased with longer incubation times. This effect could be neutralized by simulatenous incubation of the somatotropes with both regulating hormones. Hence, whereas the binding and internalization of SRIF by somatotropes do not seem to be influenced by GRH, the corresponding processes for GRH are stimulated by the presence of SRIF.
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  • 9
    ISSN: 1432-0878
    Keywords: Gills ; Indoleamines ; Immunocytochemistry ; Autonomic innervation ; Salmo gairdneri R. ; Perca fluviatilis L. ; Micropterus dolomieui (Lacépède) ; Anguilla anguilla L. ; Ictalurus melas Rafinesque (Teleostei) ; Acipenser baeri L. (Chondrostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Antibodies against serotonin and 5-methoxytryptamine reveal indolaminergic neurons innervating the proximal part of the efferent arterial vasculature, the filament epithelia, the central venous sinus, and certain other serotonergic cells of the teleost gill filament. In the same area, acetylcholinesterase-positive and indoleaminergic neurons have already been described. We propose that these populations of neurons belong to a single neuronal type but express different agents. Our current results support this idea; in particular, they point to the presence of a single type of serotonin-containing nerve terminal, impinging on vascular smooth muscle. These results are in agreement with physiological data showing (i) the existence of non-cholinergic (atropine-resistant) vasoconstriction of the gill vasculature after nerve stimulation, and (ii) a potent vasoconstrictory action of infused serotonin. In addition, the above-mentioned serotonergic neurons have synaptic contacts with catecholaminergic nerve fibers, suggesting the existence of a modulatory relationship between the sympathetic and the cranial autonomic nerves supplying the teleost gill. Finally, these neurons show morphological relationships with a previously undescribed type of branchialserotonergic cell. The role of the parasympathetic nerve plexus of the teleost gill filament in the control of respiration and ionoregulation is discussed.
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  • 10
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Secretory activity, neural control ; Transplantation ; Long-spacing collagen ; Immunocytochemistry ; Molecular markers (neuronal, glial) ; Electron microscopy ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary There is increasing evidence that, in the rat, a serotonin-mediated neural input may have an inhibitory influence on the secretory activity of the subcommissural organ (SCO). In the present investigation the rat SCO was studied 7, 30 and 90 days after transplantation under the kidney capsule, an area devoid of local serotonin-containing nerves. The grafted tissue was examined by use of immunocytochemistry employing a series of primary antisera, lectin histochemistry and transmission electron microscopy. The grafted SCO survived transplantation and contained, in addition to secretory ependymal and hypendymal SCO-cells, also elements immunoreactive with antisera against glial fibrillary acidic protein or S-100 protein. In transplants, SCO-cells produced a material displaying the characteristic immunocytochemical and lectin-binding properties of SCO-cells observed under in-situ conditions. The ependymal cells lined 1–3 small cavities, which contained secretory material. A fully developed structural equivalent of Reissner's fiber was, however, never found. The immunocytochemical and ultrastructural study of the grafted SCO showed an absence of nerve fibers within the graft and suggested a state of enhanced secretory activity. A network of protruding basal lamina structures connected the secretory cells to the newly formed capillaries revascularizing the SCO. One week after transplantation, long-spacing collagen started to appear in expanded areas of such laminar networks and also in the perivascular space. It is suggested (i) that the formation of long-spacing forms of collagen is triggered by factors provided by the SCO-secretory cells, and (ii) that secretory material of the ependymal and hypendymal cells may reach the reticular extensions of the basal lamina. In contrast to the SCO in situ, the grafted SCO-cells showed a positive immunoreaction for neuron-specific enolase. They became surrounded by a S-100-immunoreactive glial sheath that separated them from other transplanted cell types and the adjacent kidney tissue of the host.
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  • 11
    ISSN: 1432-0878
    Keywords: Gonadotropin-releasing hormone-associated peptide (GAP) ; Luteinizing hormone-releasing hormone (LHRH) ; Brain mapping ; LHRH prohormone ; Immunocytochemistry ; Rat (Wistar-R)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The gonadotropin-releasing hormone-associated peptide (GAP) of the LHRH precursor and the decapeptide LHRH were localized in the rat brain by immunocytochemistry in 12 to 18-day-old animals, by use of thick Vibratome sections and nickel intensification of the diaminobenzidinereaction product. Our results indicate that the GAP portion of the LHRH precursor is present in the same population of neurons that contain LHRH in the rat brain. An important difference observed was that the GAP antiserum, in contrast to LHRH antisera, stained several perikarya in the medial basal hypothalamus. GAP-immunoreactive perikarya were observed in the following regions: the olfactory bulb and tubercle, diagonal band of Broca, medial septum, medial preoptic and suprachiasmatic areas, anterior and lateral hypothalamus, and several regions of the hippocampus. In addition to the preoptico-terminal and the septopreoptico-infundibular pathways, we also observed GAPimmunopositive processes in several major tracts and areas of the brain, including the amygdala, stria terminalis, stria medullaris thalami, fasciculus retroflexus, stria longitudinalis medialis, periventricular plexus, periaqueductal gray of the mesencephalon and extra-cerebral regions, such as the nervus terminalis and its associated ganglion. These results confirm the specificity of previous immunocytochemical results obtained with antisera to LHRH. The presence of GAP immunoreactivity in nerve terminals of the rat brain indicates that GAP or a GAP-like peptide is located in the proper site to serve as a hypophysiotropic substance and/or as a neurotransmitter or neuromodulator.
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  • 12
    ISSN: 1432-0878
    Keywords: Thyroid gland ; Cathepsin B ; Lysosomes ; Immunocytochemistry ; Diurnal rhythm ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical localization of cathepsin B and thyroxine (T4) in follicular cells of the rat thyroid gland and plasma concentrations of thyroid stimulating hormone (TSH) were examined at six evenly spaced times over 24 h. By light- and electron microscopy, immunodeposits for cathepsin B were localized in cytoplasmic granules of various sizes, whereas those for T4 were detected mainly in larger granules of the cells and in the colloid lumen. The size and location of cytoplasmic granules showing immunoreactivity for cathepsin B and T4 in the cells varied over 24 h, corresponding to a change in plasma TSH concentrations. These immunopositive large granules appeared in the apical cytoplasm at 12.00 h, when the level of TSH was highest. At 20.00 h when the level of TSH was lowest, T4-positive granules almost disappeared, and cathepsin B-positive small granules were abundantly seen in the basal region. From 00.00 h to 08.00 h, these positive granules changed in the same manner as those seen from 12.00 h to 20.00 h, associated with an increase in plasma TSH levels. These results suggest that newly formed colloid droplets migrate from the apical to the basal regions. Cathepsin B may play a role not only in the degradation of thyroglobulin but in the maturation of thyroid hormones during the migration of the granules.
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  • 13
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    Cell & tissue research 256 (1989), S. 399-410 
    ISSN: 1432-0878
    Keywords: Dopamine ; Immunocytochemistry ; Brain, invertebrate ; Apis mellifera (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of dopamine-like immunoreactive neurons is described for the brain of the bee, Apis mellifera L., following the application of a pre-embedding technique on Vibratome sections. Immunoreactive somata are grouped into seven clusters, mainly situated in the protocerebrum. Immunoreactive interneurons have been detected in the different neuropilar compartments, except for the optic lobe neuropils. Strong immunoreactivity is found in the upper division of the central body, in parts of the stalk and in the α-lobe layers of the mushroom bodies. A dense network of many immunoreactive fibres surrounds the mushroom bodies and the central body. It forms a number of interhemispheric commissures/chiasmata, projecting partly into the contralateral mushroom body and central body. The lateral protocerebral neuropil contains some large wide-field-neurons. The antennal-lobe glomeruli receive fine projections of multiglomerular dopamine-like immunoreactive interneurons.
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  • 14
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    Cell & tissue research 257 (1989), S. 69-75 
    ISSN: 1432-0878
    Keywords: Inner ear ; Cytoskeletal proteins ; Immunocytochemistry ; Cell motility ; Actin ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Isolated outer hair cells of the guinea pig lacking a cuticular plate and its associated infracuticular network retain the ability to shorten longitudinally and become thinner. Membrane ghosts lacking cytoplasm retain the cylindrical shape of the hair-cell, and although they do not shorten, they retain the ability to constrict and become thinner. These data suggest that cytoplasmic components are associated with outer hair-cell longitudinal shortening and that the lateral wall is responsible for maintaing cell shape and for constriction. Actin, a protein associated with the cytoskeleton and cell motility, is thought to be involved in outer hair-cell motility. To study its role, actin was localized in isolated outer hair cells by use of phalloidin labeled with fluorescein and antibodies against actin coupled to colloidal gold. In permeabilized guinea-pig hair cells stained with phalloidin, actin filaments are found along the lateral wall. In frozen-fixed hair cells actin filaments are distributed uniformly throughout the cytoplasm. Electron-microscopic studies show that antibodies label actin throughout the outer hair-cell body. Thus cytoplasmic actin filaments may provide the structural basis for the contraction-like events.
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  • 15
    ISSN: 1432-0878
    Keywords: Antigen localization ; FMRFamide-like immunoreactivity ; Immunocytochemistry ; Invertebrate ganglia ; Small cardioactive peptide-like immunoreactivity ; Hirudo medicinalis (Annelida)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distributions of small cardioactive peptide (SCP)- and FMRFamide-like immunoreactivities in the central nervous system of the medicinal leech Hirudo medicinalis were studied. A subset of neurons in the segmental ganglia and brains was immunoreactive to an antibody directed against SCPB. Immunoreactive cell bodies were regionally distributed throughout the nerve cord, and occurred both as bilaterally paired and unpaired neurons. The majority of the unpaired cells displayed a tendency to alternate from side to side in adjacent ganglia. A small number of neurons were immunoreactive only in a minority of nerve cords investigated. Intracellular injections of Lucifer yellow dye and subsequent processing for immunocytochemistry revealed SCP-like immunoreactivity in heart modulatory neurons but not in heart motor neurons. FMRFamide-like immunoreactivity was also detected in cell bodies throughout the central nervous system. A subset of neurons contained both SCP- and FMRFamide-like immunoreactivities; others stained for only one or the other antigen. These data suggest that an antigen distinct from FMRFamide is responsible for at least part of the SCP-like immunoreactivity. This antigen likely bears some homology to the carboxyl terminal of SCPA and SCPB.
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  • 16
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    Cell & tissue research 256 (1989), S. 559-565 
    ISSN: 1432-0878
    Keywords: Retrograde tracing ; Immunocytochemistry ; Vascular innervation ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The origin of nerve fibers to the superficial temporal artery of the rat was studied by retrograde tracing with the fluorescent dye True Blue (TB). Application of TB to the rat superficial temporal artery labeled perikarya in the superior cervical ganglion, the otic ganglion, the sphenopalatine ganglion, the jugular-nodose ganglionic complex, and the trigeminal ganglion. The labeled perikarya were located in ipsilateral ganglia; a few neuronal somata were, in addition, seen in contralateral ganglia. Judging from the number of labeled nerve cell bodies the majority of fibers contributing to the perivascular innervation originate from the superior cervical, sphenopalatine and trigeminal ganglia. A moderate labeling was seen in the otic ganglion, whereas only few perikarya were labeled in the jugular-nodose ganglionic complex. Furthermore, TB-labeled perikarya were examined for the presence of neuropeptides. In the superior cervical ganglion, all TB-labeled nerve cell bodies contained neuropeptide Y. In the sphenopalatine and otic ganglia, the majority of the labeled perikarya were endowed with vasoactive intestinal polypeptide. In the trigeminal ganglion, the majority of the TB-labeled nerve cell bodies displayed calcitonin gene-related peptide, while a small population of the TB-labeled neuronal elements contained, in addition, substance P. In conclusion, these findings indicate that the majority of peptide-containing nerve fibers to the superficial temporal artery originate in ipsilateral cranial ganglia; a few fibers, however, may originate in contralateral ganglia.
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  • 17
    ISSN: 1432-0878
    Keywords: Insulin-related peptide ; Immunocytochemistry ; Immunochemical characterization ; Pancreas ; Ambystoma mexicanum (Urodela)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The pancreas of the axolotl, Ambystoma mexicanum, was investigated by immunocytochemical methods for the presence of immunoreactivity to a number of antisera raised against mammalian insulins. All anti-insulin antisera tested revealed substantial amounts of reaction products confined solely to the aldehyde-fuchsinophilic B cells of the endocrine pancreas. The reactive cell population was detected by use of one polyclonal antiserum against bovine insulin and eight different monoclonal antibodies against insulins from various mammalian species. Six of these antibody clones have known specificity to sub-regions of the insulin molecule. Additionally, fractions of an ethanol-HCl extract of pancreatic tissue from Ambystoma was studied in both conventional dot-blot tests by means of the same panel of antibodies and a two-site sandwich time-resolved immunofluorometric assay for human insulin involving two of the monoclonal antibodies. These experiments support the immunocytochemical observations by demonstrating the existence of an insulin-related peptide with a great deal of structural resemblance to mammalian insulins and displaying antigenic determinants in common at least with the amino acid residues A8–10 and B26–30. In conclusion, we interpret the findings as indicating that the immunocytochemically revealed tissue bound antigen in the Ambystoma pancreatic B-cells may be a peptide related to human insulin.
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  • 18
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Prolactin cells ; Gonadotropic cells ; ACTH cells ; Folliculo-stellate cells ; Rana pipiens (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The colloidal gold immunocytochemical technique was used to determine the ultrastructural features of the glandular cells in the pituitaries of male frogs, Rana pipiens, both in vivo and after superfusion in vitro. Specific reactions to antisera against bullfrog gonadotropins, human prolactin, and synthetic 1–39 corticotropin allowed identification of the 3 corresponding types of glandular cells. No immunoreaction was obtained with antisera against human or ovine-growth hormone, human β-thyrotropin hormone, and bovine S-100 protein. General morphological features of these immunocytochemically identified glandular cells were similar to those of equivalent cells previously described in other amphibian species. Non-glandular folliculo-stellate cells were distinctive. In freshly removed pituitaries, these folliculo-stellate cells contained lysosome-like structures, but did not show phagocytic vacuoles in the cytoplasm; they contained many mitochondria, and the Golgi complex and endoplasmic reticulum were relatively undeveloped. After 4 or 18 h of superfusion, some immunoreactive gonadotropic, prolactin, and corticotropic cells showed degeneration and destruction. In the same gland, folliculo-stellate cells retained a viable appearance, but showed phagocytic vacuoles containing secretory granule-like structures which were immunoreactive to gonadotropic, prolactin, and corticotropic antibodies. Some folliculo-stellate cells showed phagocytic vacuoles containing complete glandular cells. These results suggest that superfusion causes a destruction of some of the glandular cells, and that folliculo-stellate cells act as phagocytes when cellular debris or moribund cells are present in the intercellular space in the pituitary parenchyma.
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  • 19
    ISSN: 1432-0878
    Keywords: S-100 protein ; Immunocytochemistry ; Saccus vasculosus ; Pituitary gland, pars nervosa ; Mustelus manazo, Scyliorhinus torazame(Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary S-100 protein-immunoreactive cells were demonstrated by immunocytochemical procedures in the hypophysis and saccus vasculosus of two species of elasmobranchs (Mustelus manazo and Scyliorhinus torazame). In the saccus vasculosus of M. manazo, immunoreactivity was detectable exclusively in the fibrous portions interposed between the epithelial layer and the blood vessels. In the neurohypophysis, tanycytes and astrocytes of the median eminence were immunostained, but only a few labeled cells were found in the neurointermediate lobe. In S. torazame, the neurohypophysis displayed a similar distribution of immunoreactivity, but there were no labeled cells in the saccus vasculosus. In both species, none of the glandular cells of the hypophysis displayed immunoreactivity. Electron-microscopic examination showed that the immunostained cells in the saccus vasculosus correspond to astrocytes.
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  • 20
    ISSN: 1432-0878
    Keywords: Development ; Intrapulmonary chemoreceptor ; Immunocytochemistry ; Morphometry ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution, frequency and size of neuroepithelial bodies (NEB) were studied in lungs of rabbits during different stages of development (27-day fetus, newborn, 6, 11, 21, 28 and 56 days postnatally). NEB were visualized by immunostaining with monoclonal antibody against serotonin. Detailed quantitiation of NEB was performed by use of camera lucida drawings of immunostained serial sections from the same anatomical region, i.e. the lower lobe of the left lung. The total number of NEB was counted and expressed per epithelial length of airway, surface area and volume. The size of NEB defined as surface area as well as the position of NEB in relation to the airway bifurcations was assessed in airways of different sizes. The overall number and size of NEB were found to increase during the immediate perinatal period followed by a sharp decline at 56 days of age. The number of NEB peaked at 6 days postnatally (mean 175.5 NEB/mm3 of airway epithelium) and declined significantly (3.0 NEB/mm3) at 56 days of postnatal age. The size of NEB reached its maximum at 11 days (mean surface area 659.54 μm2, with the largest NEB measuring 1839.98 μm2). By 56 days of age, NEB became significantly smaller (mean surface area 177.29 μm2) consisting of small clusters of cells situated deep within the airway epithelium. At all ages, about half of all NEB (mean 47.6%) were localized within the small peripheral airways with up to 63.9% located at airway bifurcations. These findings indicate that the “functional activity” of NEB may be confined predominantly to the perinatal period. The postulated functions of NEB include those of intrapulmonary hypoxia-sensitive chemoreceptors and/or endocrine-paracrine activity in the lung. Such function(s) may be important during adaptation to extrauterine life as well as for growth and development of the lung.
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  • 21
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    Cell & tissue research 256 (1989), S. 45-51 
    ISSN: 1432-0878
    Keywords: Testis ; Leydig cells ; Basement membrane ; Laminin ; Collagen ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Rat testes were examined by conventional and immunolabeling transmission electron microscopy. Ultrastructurally identifiable continuous basement membranes were found around seminiferous tubules and the interstitial capillaries. Patches of basement membrane were, additionally, found on free surfaces of Leydig cells, between two Leydig cells, and in macrophage-Leydig cell contact sites. The ultrastructural findings were confirmed by immunocytochemical localization of laminin and collagen type IV in the same areas. A close association between the capillary basement membranes and the surfaces of perivascular Leydig cells was also observed. The possible basement membrane-mediated interactions of Leydig cells with other testicular structures, together with the novel bioactive products and regulators of Leydig cells, support the role of these cells as exceptionally complex regulatory centers of testicular functions.
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  • 22
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    Cell & tissue research 256 (1989), S. 153-158 
    ISSN: 1432-0878
    Keywords: Acetylcholine ; Acetylcholinesterase ; Cholin-acetyltransferase ; Immunocytochemistry ; Visual system ; Calliphora erythrocephala (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of putative cholinergic neurons in the lamina of the blowfly Calliphora erythrocephala was studied by immunocytochemical and histochemical methods. Three different antibodies directed against the AChsynthesizing enzyme, choline acetyltransferase (ChAT), revealed a cholinergic population of fibres running parallel to the laminar cartridges, which have branch-like structures at the distal lamina border. Cell bodies in the chiasma next to the lamina border were also labelled by the anti-ChAT antibodies. Monopolar cell bodies in the nuclear layer were faintly labelled. The distribution of the acetylcholine hydrolyzing enzyme, acetylcholine esterase (AChE), was revealed by histochemical staining and was similar to the ChAT immunocytochemistry. The arrangement of ChAT positive fibres in transverse and longitudinal sections and the distribution of AChE stained fibres indicate that the amacrine cells of the lamina are cholinergic cells.
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  • 23
    ISSN: 1432-0878
    Keywords: Pancreas, endocrine ; Cysteamine ; Somatostatin ; Insulin secretion ; Glucose ; Immunocytochemistry ; Mouse (NMRI)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary To evaluate the previously reported depletion of pancreatic somatostatin by cysteamine (β-mercaptoethylamine), mice were injected subcutaneously with the drug at 300 mg/kg. Immunocytochemical analysis performed on sections from tissue taken at 4 h after the injection revealed an elimination of somatostatin-14-like immunoreactivity without alterations in the somatostatin-28(1 – 12)-like immunoreactivity. In sections from tissues taken at 24 h after injection, no differences between cysteamine-injected animals and controls were observed. Immunochemical analysis of somatostatin-14-like immunoreactivity in pancreatic extracts showed a significant reduction of the concentration (P〈 0.001). In contrast, no change in the insulin concentration was observed. Functionally, cysteamine lowered the plasma glucose levels at l h after injection; this effect persisted for 6 h. Plasma insulin levels were likewise reduced transiently by cysteamine. Concomitant administration of somatostatin did not influence these effects of cysteamine. The plasma glucose-lowering effect of cysteamine was seen also in alloxan-diabetic mice. We conclude that cysteamine alters the immunoreactive characteristics of pancreatic somatostatin without affecting the immunoreactivity of insulin, and that cysteamine transiently reduces plasma glucose and insulin levels
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  • 24
    ISSN: 1432-0878
    Keywords: FMRFamide ; Cholecystokinin ; Immunocytochemistry ; Insect visual system ; Neural development ; Calliphora erythrocephala (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The adult optic lobes of the blowfly Calliphora erythrocephala were found to be innervated by more than 2000 neurons immunoreactive to antisera raised against the neuropeptides FMRFamide, its fragment RFamide, and gastrin/cholecystokinin (CCK). All of the CCK-like immunoreactive (CCK-IR) neurons also reacted with antisera to RFamide, FMRFamide and pancreatic polypeptide. A few RFamide/FMRFamide-like immunoreactive (RF-IR) neurons did not react with CCK antisera; they reacted instead with antisera to Leu-enkephalin and Met-enkephalin-Arg6-Phe7. The RF-IR neurons are, thus, heterogeneous with respect to their contents of immunoreactive peptides. Two of the RF-IR neuron types innervating the adult optic lobes could be traced in their entirety only after following their postembryonic development, because of the complexity of the trajectories of the immunoreactive neuronal process in the adult insect. The majority of the cell bodies of the RF-IR and CCK-IR neurons lie within the optic lobes and are derived from imaginal neuroblasts of the inner and outer optic anlagen. Six of the peptidergic neurons are, however, metamorphosing larval neurons with their cell bodies in the central part of the protocerebrum. The full extent of immunoreactivitiy is not attained in some of the neurons until the late pupal or early adult stage. The larval optic center was also found to be innervated by neurons immuno-reactive with both RFamide and CCK antisera. The cell bodies of these RF-IR/CCK-IR neurons are located near the developing lamina (one on each side). In the 24 h pupa, the cell bodies of these neurons are still immunoreactive, but thereafter they cannot be immunolabeled apparently due to cell death or a change in transmitter phenotype.
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  • 25
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    Cell & tissue research 255 (1989), S. 645-650 
    ISSN: 1432-0878
    Keywords: Pituitary, pars distalis ; Differentiation ; Organ culture ; Immunocytochemistry ; Rat (Sprague-Dawley, CFY)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An immunohistochemical study was performed to determine the capacity of early fetal pituitaries to differentiate into specific hormone-synthesizing tissue in the absence of any influence from the central nervous system. Rathke's pouches from rats were removed from their juxtadiencephalic position on day 11 and 12 of gestation and maintained for 2–7 days in a chemically defined culture medium (M 199) without antibiotics and serum supplementation. The immunocytochemical observations provided evidence for the differentiation of ACTH-, TSH-gb-, LH-gb-, FSH-gb-, GH- and PRL-synthesizing cells in the isolated organ cultured from 11 to 12-day-old pituitary primordia. The appearance of specific hormone-synthesizing cells in vitro displayed a delay of 1.5–2 days compared to the day of appearance in vivo, however, the sequential order of developmental events occurred as observed in vivo. The present results suggest that endocrine or neuroendocrine signals are not required for the expression of specific secretory functions of fetal pituitaries, at least at an age of 11–12 days.
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  • 26
    ISSN: 1432-0878
    Keywords: Hypophyseal pars tuberalis ; (TSH), Thyrotropin ; Immunocytochemistry ; Photoperiod ; Phodopus sungorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Specific secretory cells in the hypophyseal pars tuberalis of Djungarian hamsters maintained under different photoperiods were investigated immunocytochemically by means of the colloidal gold technique using antibodies against rat thyrotropin (TSH). Secretory cells of animals kept under long photoperiods (LD16:8) showed positive staining of secretory granules (diameters 90–130 nm), whereas other intracellular structures were free of immunoreactivity. In animals kept under short photoperiods (LD8:16) secretory cells displayed increased numbers of secretory granules, but these organelles were devoid of immunoreactivity. In contrast, immunoreactivity of thyrotropes in the pars distalis did not differ between the two groups of animals investigated. The present results confirm earlier light-microscopical studies that in the pars tuberalis specific secretory cells show TSH-like immunoreactivity; however, they differ in their reactivity pattern from classical thyrotropes in the pars distalis.
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  • 27
    ISSN: 1432-0878
    Keywords: Neuropeptides ; Monoclonal antibodies ; Immunocytochemistry ; Endocrine system ; Nervous system ; Leptinotarsa decemlineata (Say)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Monoclonal antibodies were developed against peptidergic neurons in the nervous and endocrine tissues of the Colorado potato beetle by the immunization of mice with unpurified homogenates of these tissues. Methods were optimized to enhance chances for successful antibody production and selection, such as the pretreatment of the beetles, preparation of the immunogen, and screening hybridomas. Although only sub-microgram quantities of peptide antigen were used, many hybridomas generated antibodies recognizing peptidergic neurons in immunocytochemical procedures. A panel of 13 monoclonal antibodies anti-Colorado potato beetle (MACs) were harvested. All MACs stained different populations of peptidergic neurons, some of which had not been revealed by previously applied identification methods. Apart from the intrinsic glandular cells in the corpora cardiaca, immunoreactive neurosecretory neurons were revealed in medial and lateral groups in the protocerebrum and in the suboesophageal ganglion. These have axons terminating in the corpora cardiaca, and the neurosecretory granules can be revealed with the immunogold method. It is suggested that the immunoreactive substances represent neuropeptides or precursors of different kinds. Interneurons in other locations in the central and visceral nervous system have immunoreactive axonal projections that do not leave the ganglia. The set of MACs obtained is useful for neuroanatomical studies, for characterizing the secretory products, and for a further delineation of peptidergic communication channels in the insect body.
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  • 28
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    Cell & tissue research 255 (1989), S. 215-224 
    ISSN: 1432-0878
    Keywords: Transglutaminase ; Immunocytochemistry ; Induction ; Tissue compartments ; Tissue integrity ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Tissue transglutaminase is an intracellular enzyme without established physiological function. Biochemically it can be detected in all organs, but no systematic in situ localization has been carried out so far. Here we report the immunohistochemical localization of transglutaminase in human tissues using an affinity purified, monospecific anti-human transglutaminase antibody. It is shown that the widespread organ distribution of the enzyme is the consequence of its occurrence in ubiquitous cell types such as endothelium and smooth muscle cells. Some organ-specific cell types express the enzyme constitutively (mesangial cells, renomedullary interstitial cells, thymic subcapsular epithelium, colonic pericryptal fibroblasts), while in others it seems to be induced either by external stimuli (epithelium of the female breast) or as part of their differentiation/maturation program (developing nephrons, enterocytes of the small intestine). The presence of tissue transglutaminase can be demonstrated in derivatives of all germ layers and in the trophoblast. The functional implications of these findings are presently unknown; however, based on its distribution the role of this enzyme in compartmentation and preservation of tissue integrity against stress may be suggested.
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  • 29
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Neural lobe ; Intermediate lobe ; Intraventricular graft ; Immunocytochemistry ; Electron microscopy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neurointermediate lobes from adult or 10-dayold rats were implanted by a stereotaxic procedure into the third ventricle of adult male rats, in an area close to the paraventricular nucleus. They were examined, using immunocytochemical and ultrastructural techniques, at times ranging from 1 week to 8 months. All grafts were recovered in a healthy condition although some rejection of the tissue was detected at the 1and 2-week stages. In the neural lobe, clusters of pituicytes were scattered among the loose network of capillaries, most of which had a fenestrated endothelium. The intermediate lobe remained organized in compact avascular lobules. Axons similar to those projecting into the neurointermediate lobe in situ, but also axons of other types (e.g., somatostatinergic, enkephalinergic) penetrated the grafts. Synapses with melanotrophic cells in the intermediate lobe and neurohaemal contacts in the neural lobe were frequent from 2 1/2 months after transplantation. Immunocytochemical and ultrastructural characteristics indicated intense secretory stimulation of the melanotrophic cells in the early stages. All cells enclosed in a same glandular lobule reacted in a similar manner. In later stages, when re-innervation occurred, the cells recovered their initial characteristics. The overall effect of the re-innervation of the intermediate lobe grafted in this location is inhibitory, as in the lobe in situ.
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  • 30
    ISSN: 1432-0878
    Keywords: Spinal cord ; Dorsal root ganglia ; Skin ; Immunocytochemistry ; Neuropeptides ; Mutilated foot rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The mutilated foot rat is a mutant with autosomal recessive sensory neuropathy and frequent mutilation of the hindlimbs. Decreased numbers of dorsal root ganglion cells and diminished sensitivity to painful stimuli are characteristics of these animals. By use of immunocytochemistry, changes in the distributions of peptides involved in sensory and/or autonomic regulation, i.e. calcitonin generelated peptide (CGRP), tachykinins, enkephalin and neuropeptide Y in spinal cord, dorsal root ganglia and skin of these animals, were studied. In comparison with normal litter-mate controls, the dorsal horn of mutilated foot rats contained substantially fewer CGRP and tachykinin-immunoreactive fibres but more fibres immunoreactive for enkephalin. Many enkephalin-immunoreactive cell bodies were also found in the dorsal horn of the mutants, by contrast none were visible in control animals. Neuropeptide Y immunoreactivity was, however, unchanged in the spinal cord of the mutants. In the dorsal root ganglia of the mutants, the number of CGRPor tachykinin-immunoreactive cells and their proportion to total neuronal numbers were significantly less in comparison with normal controls. The diameter range of CGRP- and tachykinin-immunoreactive cells shifted from small (15–25 μm) to medium size (25–45 μm) as revealed by frequency distribution histograms. The skin from the affected foreand hindlimbs of the mutant rats, in keeping with fewer CGRP- and tachykinin-immunoreactive cells in the dorsal root ganglia, contained substantially less fibres immunoreactive for CGRP and tachykinins; a difference that was not seen in skin of unaffected areas (whiskers and snout). By contrast, neuropeptide Y-immunoreactive fibres showed a normal distribution around blood vessels and sweat glands of mutilated foot rats. The data suggest that diminished pain perception in the mutilated foot rat is related to loss of peptide-containing sensory neurones. Furthermore, the intraspinal increase of enkephalinergic neurones in the dorsal horn, concomitant with the decreased number of primary sensory neurones, may also play a contributory rôle in reducing pain thresholds.
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  • 31
    ISSN: 1432-0878
    Keywords: Cell proliferation ; Immunocytochemistry ; Lung ; Bronchioles ; Alveoli, lung ; Proliferating cell nuclear antigen ; Type II pneumocyte ; Clara cell ; Mouse (various strains)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Proliferating cell nuclear antigen is expressed in cells from late G1 through the S-phase of the cell cycle. Therefore, antibodies directed against this molecule should provide a probe for labeling immunocytochemically the nuclei of proliferating cells. Herein we demonstrate the feasibility and reliability of this technique by quantifying immunostained pulmonary nuclei. We applied polyclonal and monoclonal antisera to alveolar and bronchiolar pulmonary epithelial cells in various proliferative states in tissue-sections and in vitro. A/J mice had a slightly higher labeling index than C57BL/6J mice, and proliferation in both strains increased dramatically after butylated hydroxytoluene treatment produced compensatory hyperplasia of Type-II pneumocytes. Immunostaining in fetal and neonatal lung samples from mice was higher than in adults. Spontaneous lung adenomas had a higher labeling index than the surrounding normal lung tissue. In addition, new data contained herein demonstrate a strain difference in proliferation of bronchiolar epithelial cells, and quantify the extent to which BHT-induced lung damage increases these proliferative rates. This mammalian nuclear antigen did not cross-react with antiserum to a functionally related bacterial protein, the beta subunit of E. coli DNA polymerase-III holoenzyme.
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  • 32
    ISSN: 1432-0878
    Keywords: Pars distalis ; Prolactin/thyrotropic cells ; TRH ; Immunocytochemistry ; Morphometry ; Rana perezi (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effects of synthetic thyrotropin-releasing hormone on pituitary prolactin and thyrotropic cells were investigated in adult male Rana perezi (formerly Rana ridibunda) frogs. Animals were given daily injections of synthetic thyrotropin-releasing hormone into the dorsal lymph sac. Prolactin and thyrotropic cells were identified by the colloidal-gold method, using anti-human prolactin and anti-human-β-thyrotropin hormone as primary antisera. The stereological parameters of the rough endoplasmic reticulum, Golgi complex, and secretory granules of prolactin and thyrotropic cells were evaluated by ultrastructural morphometry (point-counting method). Thyrotropin-releasing hormone caused cytological changes in both cell-types which were consistent with increased synthesis and release of both prolactin and thryrotropin. These changes were still significant after 48 h treatment in the case of thyrotropic cells, while in prolactin cells the thyrotropin-releasing hormone increased the number of secretory granules. After 6 days, the cells resembled essentially those used as controls. These results indicate that thyrotropin-releasing hormone stimulates the synthesis and release of prolactin and thyrotropin, and that the response of each cell type to this hypothalamic stimulus follows a different time-course.
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  • 33
    ISSN: 1432-0878
    Keywords: Peptides ; Small intensely fluorescent (SIF) cell ; Neuropeptide coexistence ; Inferior mesenteric ganglion ; Immunocytochemistry ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Coexistence of peptides in the small intensely fluorescent cells was demonstrated by immunocytochemistry for met-enkephalin-Arg-Gly-Leu, vasoactive intestinal polypeptide, somatostatin, neuropeptide Y and dynorphin. In the extreme example, a single cell was immunoreactive to all 5 peptides examined. Four peptides coexisted in 8% and three peptides in 13% of SIF cells. In 10% of SIF cells no peptide immunoreactivity could be detected. The most prevalent peptide was met-enkephalin (in 46% of cells), then vasoactive intestinal polypeptide (45%), somatostatin (39%), neuropeptide Y (31%) and dynorphin (24%). Met-enkephalin and vasoactive intestinal polypeptide coexisted most commonly (25%).
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  • 34
    ISSN: 1432-0878
    Keywords: Endocrine heart ; Estradiol ; Autoradiography ; Immunocytochemistry ; Co-localization ; CDD/ANP gene regulation ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Atrial myoendocrine cells of rat were investigated regarding estradiol uptake. It was found that, in addition to their specific endocrine function of producing cardiac polypeptides of the cardiodilatin/atrial natriuretic peptide (CDD/ANP) family, these cells also specifically accumulate radiolabeled estradiol. This co-localization supports the view that steroid hormones play an important role in the regulation of the CDD/ANP gene.
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  • 35
    ISSN: 1432-0878
    Keywords: Prolactin cells ; Growth hormone cells ; In situ hybridization ; Immunocytochemistry ; Cytogenesis ; Rat (Wistar-Imamichi)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cytogenesis of growth hormone and prolactin cells in the rat pituitary gland was studied using in situ cDNA-mRNA hybridization and immunocytochemistry. Frozen or Paraplast sections of fetal and neonatal pituitaries were hybridized with 3H-cDNAs for rat prolactin or growth hormone, and were then processed for autoradiography. A number of growth hormone mRNA-positive cells were encountered throughout the anterior lobe on day 19 of gestation. Individual variaction in growth hormone gene expression was observed between fetuses at day 19 of gestation (6 out of 8 fetuses examined were positive for growth hormone mRNA). In contrast, growth hormone mRNA was detected in the all fetuses examined on day 20 or later. The autoradiographic signal (number of reduced silver grains) appeared to increase with later stages of development. Fetal growth hormone mRNA-positive cells were evenly scattered throughout the anterior lobe. Most of them were isolated, however, small clusters of several growth hormone cells were infrequently observed. Prolactin mRNApositive cells were found first on the 22nd day (the last day of gestation) in 3 of 6 fetuses examined, but were rarely observed on earlier gestational days. By postnatal day 8, prolactin mRNA-positive cells were numerous and the grain density over prolactin cells increased. Both growth hormone and prolactin cells were found as early as 18 days of gestation using immunocytochemistry, although the number of positive cells was very small at this stage. Immunoreactive growth hormone cells increased sharply in number during the next 24 h, while the number of prolactin cells remained scarce until birth. The results suggest that many growth hormone cells are still in an immature state at 20∶00 of day 18 and that many begin to synthesize growth hormone mRNA during next 14 h. On the other hand, no substantial prolactin gene expression appears to take place until after birth.
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  • 36
    ISSN: 1432-0878
    Keywords: Nucleus intercollicularis ; α2-Adrenergic receptors ; Estrogen receptors ; Quantitative autoradiography ; Immunocytochemistry ; Japanese quail, Coturnix coturnix japonica (Aves, Phasianiformes)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The nucleus intercollicularis is an important site in the control of vocalization in birds. In oscines, a subregion of the nucleus intercollicularis called the dorso-medial intercollicular nucleus appears to play a key role in this process because it receives the majority of the projections from the nucleus robustus archistriatalis and sends most of the projections to the motor nucleus of the hypoglossal nerve. In this paper, we present neurochemical studies of the nucleus intercollicularis in the Japanese quail which suggest the presence of heterogeneity within this structure. One rostral band contains high densities of cholinergic muscarinic receptors identified by quantitative autoradiography using tritiated N-methylscopolamine as the ligand. A caudal dorso-medial region is specifically labeled by estrogen receptors identified using immunocytochemistry and by α2-adrenergic receptors which were quantified by autoradiography using tritiated para-amino-clonidine. This latter sub-region is possibly equivalent to the dorso-medial intercollicular nucleus of oscines. Additional track-tracing studies should be performed to confirm this homology. The coexistence of estrogen and α2-adrenergic receptors within the same structure suggests important functional connections between steroid action and catecholaminergic systems in the brain.
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  • 37
    ISSN: 1432-0878
    Keywords: Thyrotropin-releasing hormone (TRH) ; Development, ontogenetic ; Anterior hypothalamus ; Immunocytochemistry ; Domestic mallard
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Developmental changes of thyrotropin-releasing hormone (TRH)-immunoreactive structures in the brain of mallard embryos were studied by means of immunocytochemistry (PAP technique). The primary antibody was generated against synthetic TRH. Immunoreactive neurons were first detected in the hypothalamus of 14-day-old embryos. By day 20, increasing numbers of immunoreactive perikarya were observed in the paraventricular nucleus, anterior preoptic region and supraoptic region. Immunoreactive fiber projections were seen in the median eminence as early as embryonic day 20; they occurred also in some extrahypothalamic regions (lateral septum, accumbens nucleus). The number and staining intensity of the cell bodies increased up to hatching, and continued to increase during the first week after hatching.
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  • 38
    ISSN: 1432-0878
    Keywords: Olfactory-marker protein (OMP) ; Olfactory epithelium ; Immunocytochemistry ; Protein G-gold ; Freeze-substitution ; Lowicryl K11M embedding ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In this study electron-microscopic immunocytochemistry was used to localize olfactory marker protein in olfactory epithelia. Rat olfactory-epithelial samples were rapidly frozen, freeze-substituted with acetone, embedded at low temperatures with Lowicryl K11M and labelled on the sections with polyclonal antibodies raised against olfactory marker protein and with protein G conjugated to colloidal gold. Apart from the aforementioned use of acetone, substitution was carried out in the complete absence of chemical fixation, i.e., neither aldehydes nor OsO4 were used. This procedure resulted in localization concurrent with a good ultrastructural preservation. Olfactory-marker protein was present throughout the cytoplasmic compartments of dendrites and dendritic endings of olfactory-receptor cells, but it was not found in organelles such as mitochondria. Olfactory-marker protein was found only in dendriticendings of olfactory-receptor cells mature enough to have given rise to cilia, but these cilia displayed less labelling than dendrites and dendritic endings. Olfactory-marker protein was not found in apices and microvilli of neighboring olfactory-supporting cells.
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  • 39
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Glial markers ; Immunocytochemistry ; GABA uptake ; Comparative analysis ; Mammals (rat, cat, mouse, rabbit)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subcommissural organ (SCO), classified as one of the circumventricular organs, is composed mainly of modified ependymal cells, attributable to a glial lineage. Nevertheless, in the rat, these cells do not possess glial markers such as glial fibrillary acidic protein (GFAP), protein S100, or the enzyme glutamine synthetase (GS). They receive a synaptic 5-HT input and show pharmacological properties for uptake of GABA resembling the uptake mechanism of neurons. In this study, we examine the phenotype of several mammalian SCO (cat, mouse, rabbit) and compare them with the corresponding features of the rat SCO. In all these species, the SCO ependymocytes possess vimentin as an intermediate filament, but never express GFAP or neurofilament proteins. They do not contain GS as do glial cells involved in GABA metabolism, and when they contain protein S100 (rabbit, mouse), its rate is low in comparison to classical glial or ependymal cells. Thus, these ependymocytes display characteristics that differentiate them from other types of glial cells (astrocytes, epithelial ependymocytes and tanycytes). Striking interspecies differences in the capacity of SCO-ependymocytes for uptake of GABA might be related to their innervation and suggest a species-dependent plasticity in their function.
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  • 40
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    Cell & tissue research 257 (1989), S. 343-366 
    ISSN: 1432-0878
    Keywords: Central complex ; Golgi impregnation ; Neurotransmitters ; Protocerebrum, insect ; Immunocytochemistry ; Drosophila melanogaster (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary On the basis of 1200 Golgi-impregnated brains the structure of the central complex of Drosophila melanogaster was analyzed at the cellular level. The four substructures of the central complex — the ellipsoid body, the fanshaped body, the noduli, and the protocerebral bridge — are composed of (a) columnar small-field elements linking different substructures or regions in the same substructure and (b) tangential large-field neurons forming strata perpendicular to the columns. At least some small-field neurons belong to isomorphic sets, which follow various regular projection patterns. Assuming that the blebs of a neuron are presynaptic and the spines are postsynaptic, the Golgi preparations indicate that small-field neurons projecting to the ventral bodies (accessory area) are the main output from the central complex and that its main input is through the large-field neurons. These in turn are presumed to receive input in various neuropils of the brain including the ventral bodies. Transmitters can be attributed immunocytochemically to some neuron types. For example, GABA is confined to the R1–R4 neurons of the ellipsoid body, whereas these cells are devoid of choline acetyltransferase-like immunore-activity. It is proposed that the central complex is an elaboration of the interhemispheric commissure serving the fast exchange of data between the two brain hemispheres in the control of behavioral activity.
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  • 41
    ISSN: 1432-0878
    Keywords: L1-antigen ; Cell adhesion molecule ; Developing vestibular neuroepithelium ; Immunocytochemistry ; Mouse (CBAxC57)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Doubts exist as to whether afferent nerve fibers exert a neurotrophic effect on the differentiation of sensory cells in the developing vestibular neuroepithelium. To determine whether innervation of hair cells precedes their differentiation, we have used the L1 adhesion molecule as a marker for axons. The detection of L1 on afferent axons in the otic vesicle of mouse embryos on gestation day 11 shows that nerve fibers penetrate the neuroepithelium before the sensory cells differentiate. L1-immunoreactivity of nerve endings also reveals the considerable fiber ramification on gestation days 14 and 15, i.e., corresponding to the first stages of sensory cell differentiation. The expression of L1 at successive stages of nerve fiber growth in the neuroepithelium, such as fasciculation and ramification, is not consistent with the previous role proposed for L1 as a fascicule-promoting factor and raises the possibility that other mechanisms are involved in L1 mediaded adhesion.
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  • 42
    ISSN: 1432-0878
    Keywords: Neuropeptides ; Immunocytochemistry ; Lumbricus terrestris (Annelida, Oligochaeta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localisation and distribution of 10 vertebrate-derived neuropeptides in the earthworm, Lumbricus terrestris, have been determined by an indirect immunofluorescence technique. The peptides are pancreatic polypeptide (PP), peptide tyrosine tyrosine (PYY), neuropeptide Y (NPY), glucagon (C-terminal), vasoactive intestinal polypeptide (VIP), peptide histidine isoleucine (PHI), gastrinreleasing peptide (GRP), calcitonin gene-related peptide (CGRP), neurotensin (NT), and met-enkephalin. For 6 of the peptides — PYY, NPY, PHI, glucagon, GRP and CGRP — this is the first demonstration of their presence in any annelid, and NT has not previously been described in an oligochaete. Cell bodies and nerve fibres immunoreactive to the 10 peptides occur throughout the CNS. In the PNS, epidermal sensory cells displayed immunoreactivities to PP and PYY, and PP-, PYY-, NPY-, PHI- and GRP-like immunoreactivities occurred in nerve fibres supplying the main body muscles. Nerve fibres immunoreactive to PP and PYY are also associated with the innervation of the gut (pharynx, oesophageal glands, and mid and posterior regions of the intestine). No endocrine cells immunoreactive for any of the antisera tested could be identified in the gut epithelium, suggesting that dual location of peptides in the brain and gut epithelium is a phenomenon that occurred at a later stage in evolution. No immunoreactive elements were detected in any of the organs and ducts of the reproductive and excretory systems.
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  • 43
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 255 (1989), S. 225-233 
    ISSN: 1432-0878
    Keywords: GABA ; Immunocytochemistry ; Visual system ; Optic lobes ; Periplaneta americana (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Specific antisera against protein-conjugated γ-aminobutyric acid (GABA) were used in immunocytochemical staining procedures to study the distribution of the putative GABA-like immunoreactive neurons in the optic lobes of Periplaneta. GABA-like immunoreactive structures are evident in all three optic neuropil regions. Six different populations of GABAergic neurons, whose perikarya are grouped around the medulla, are found within the optic lobe. The number of these immunoreactive cells varies greatly and corresponds to the number of ommatidia of the eye. In the proximal part of the lamina, a coarse network of GABA-positive fibres is recognizable. These are the processes of large field tangential cells whose fibres pass through the distal surface of the medulla. A second fibre population of the lamina is made up of the processes of the centrifugal columnar neurons whose perikarya lie proximally to the medulla. The medulla contains 9 layers with GABAergic elements of variable immunoreactivity. Layers 1, 3, 5, 7 and 9 exhibit strong labelling, as a result of partial overlapping of the processes of centrifugal and centripetal columnar neurons, tangential fibres and/or lateral processes of perpendicular fibres and (possibly) processes of amacrines. A strong immunoreactivity is found in the proximal and distal layers of the lobula.
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  • 44
    ISSN: 1432-0878
    Keywords: Serotonin (5HT) ; Immunocytochemistry ; 5,6-Dihydroxytryptamine ; Central nervous system ; Helix pomatia (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of serotonin (5HT)-containing neurons in the central nervous system of the snail Helix pomatia has been determined in whole-mount preparations by use of immunocytochemical and in vivo 5,6-dihydroxy-tryptamine labelling. 5HT-immunoreactive neuronal somata occur in all but the buccal and pleural ganglia. Immunoreactive fibres are present throughout the central nervous system. The 5HT-immunoreactive neuronal somata characteristically appear in groups, located mainly in the cerebral, pedal, visceral and right parietal ganglia. The majority of 5HT-immunoreactive neurons is located in the pedal ganglia. Additionally a dense network of 5HT-immunoreactive varicose fibres is found in the neural sheath of the central nervous system including all the nerves and ganglia. The number and distribution of 5HT-immunoreactive neurons correlates with that demonstrated by 5,6-dihydroxytryptamine labelling method.
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  • 45
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 257 (1989), S. 545-548 
    ISSN: 1432-0878
    Keywords: Brush cells ; Fatty acid-binding protein ; Immunocytochemistry ; Stomach ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An immunocytochemical study by light- and electron microscopy using the antibody against rat hepatic fatty acid-binding protein (FABP) revealed the brush cells in the gastric epithelium of rats to be intensely immunoreactive. The immunoreactive cells were present in a group in the distal wall of the groove between forestomach and glandular stomach, as well as scattered singly in the surface and foveolar epithelia of the glandular stomach. Almost all immunoreactive brush cells had a thin basal process in contact with the basement membrane. No secretory granules with dense cores, similar to those of endocrine cells, were observed in the brush cells. The specific appearance of FABP-immunoreactivity in the brush cell indicates that this cell type is a distinct entity from other epithelial cells in the stomach and that FABP is a useful histochemical marker of the brush cells. FABP may be involved in the specific function(s) of this cell type related to fatty acid metabolism.
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  • 46
    ISSN: 1615-6102
    Keywords: Pyrenoid ; Ribulose 1,5-bisphosphate carboxylase/oxygenase ; Chlorella pyrenoidosa ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The localization of ribulose 1,5-bisphosphate carboxylase/ oxygenase (RuBisCO) in cells ofC. pyrenoidosa grown at varying light intensities was determined by immunoelectron microscopy. Log phase cells grown at photon flux densities of 25 and 75 μEm−2s−1 (light-limiting) and 540 μEm−2s−1 (light-saturating) were fixed in 3% glutaraldehyde and embedded in Lowicryl K4M. Sections were labelled with antiserum to each subunit of RuBisCO followed by protein A-gold. At each light fluence rate, the pyrenoid was heavily labelled by each antibody whereas chloroplast stromal labelling was not above background levels. The apparent absence of stromal RuBisCO at each light level, and hence the lack of enzyme redistribution from pyrenoid to stroma following an increase in light fluence rate, suggests that pyrenoid RuBisCO is functional in vivo.
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