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  • 1
    ISSN: 0196-9781
    Keywords: Capsaicin ; Dorsal root ganglia ; Immunocytochemistry ; Lateral dorsal horn ; Lissauer's tract ; Lumbar spinal cord ; Pelvic ganglion ; Pelvic nerve section ; Peptide histidine isoleucine ; Radioimmunoassay ; Retrograde tracing ; Sacral spinal cord ; Vasoactive intestinal polypeptide
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0196-9781
    Keywords: Denervation ; Ganglia ; Immunocytochemistry ; Radioimmunoassay ; Rat ; Urinary bladder ; Vasoactive intestinal polypeptide
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A quantitative immunohistochemical study was performed of the distribution of protein gene product 9.5 (PGP, a soluble protein localized in neurons and neuroendocrine cells as well as in some non-nervous cells) and ubiquitin along the rat epididymis. In the ductuli efferentes, PGP immunoreaction was observed in the whole cytoplasm of some columnar cells; a smaller number of columnar cells showed ubiquitin immunoreactivity with limited apical and basal cytoplasmic localization. In the proximal caput epididymidis, the whole cytoplasm of all columnar cells showed PGP immunoreactivity, ubiquitin immunostaining was negative in this region. In the middle and distal caput epididymidis and the distal cauda, the apical cytoplasm of some columnar cells and the whole cytoplasm of some basal cells showed immunoreactivity to PGP. In these regions, immunoreactivity to ubiquitin was positive in the supranuclear cytoplasm of some columnar cells but not in the basal cells. No immunoreactivity to PGP or ubiquitin was detected in the corpus epididymis and the proximal cauda. Double immunostaining revealed that all the epididymal ubiquitin immunoreactive cells were also PGP immunoreactive, whereas most PGP immunoreactive cells did not immunoreact to ubiquitin. In ubiquitin-PGP immunoreactive cells, the site of the PGP immunoreaction differed from that of the ubiquitin immunoreaction. PGP-ubiquitin immunoreactive cells also seemed to be immunoreactive to anti-AE1/AE3 keratin antibodies. The spermatozoal heads were immunoreactive to PGP antibodies in the epididymal regions from proximal caput to distal cauda but not in the ductuli efferentes. The findings suggest that non-ubiquitinated PGP immunoreactive proteins are secreted in the epididymis, mainly in the proximal caput, and attach to spermatozoa.
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  • 4
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary VIP- and substance P-like immunoreactivities were found in considerable concentrations (VIP: 17.3±4.8 pmol/g, mean ± SEM; substance P:11.1±1.8 pmol/g) in the uveal portion of the guinea pig eye.d Immunocytochemistry localised these two regulatory peptides to nerve fibres found principally in a plexus in the iris (substance P) and in an extensive network surrounding the blood vessels of the choroid (VIP). A remarkable anatomical demarcation of the two types of peptide-containing nerves was established by the staining of substance P-containing nerves, which stops at the level of the ciliary body. This uveal area is known to be involved in the ocular responses to nociceptive stimuli. At the ultrastructural level, immunoreactivity for both peptides was localised to distinct subpopulations of p-type nerves, distinguishable by the size of their large dense-cored vesicles. Those immunoreactive for VIP were significantly larger (p〈0.0005) than those immunoreactive for substance P (95±7 nm and 82±9 nm respectively; mean ± SD). Interruption of the trigeminal pathway produced a remarkable decrease of substance P immunoreactivity in the anterior portion of the uvea (9.1±1.5 pmol/g, mean ± SEM, control; 5.3±1.3 pmol/g, denervated), but not of VIP immunoreactivity in the choroid. Following colchicine treatment, VIP-immunoreactive neuronal cell bodies were localised in the choroid. The separate anatomical localisations and distributions of the two uveal peptides appear to be related to their different origins and functional roles in the response of the eye to noxious stimuli.
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  • 5
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The morphological relationship between sensory and sympathetic nerves was studied in tissues of the eye and the oral cavity following chronic sympathetic or sensory denervation. Immunoreactivities for calcitonin gene-related peptide (CGRP) and tyrosine hydroxylase (TH) were used as indexes to assess the changes of the two nerve populations after denervation. Following surgical sympathectomy, a marked increase of CGRP-containing fibres was seen in all tissues studied, while TH-imunoreactive fibres were totally depleated. Conversely, after capsaicin treatment, an increase of TH-immunoreactive nerves was found in the same tissues, concomitant with a sharp decrease of CGRP-immunoreactive nerves. These changes were particularly evident in iridial stroma and around blood vessels in all tissue, where sensory and sympathetic nerves have a closely overlapping distribution pattern. The altered proportion of sensory peptide-and catecholamine-containing nerves following sympathetic and sensory denervation suggest that there is a reciprocal trophic influence between the two nerve subsets, possibly with the intervention of neurotrophic substances such as nerve growth factor. These results indicate a close interaction between sensory peptidergic and sympathetic nervous systems in peripheral organs.
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  • 6
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Several studies have shown the use of non-radioactive labelled DNA probes for in situ hybridisation, mainly to identify cellular DNA. In this study mRNA in situ hybridisation was performed on rat pituitary with biotinylated complementary (c) RNA probes for rat prolactin and growth hormone (GH), and compared with radioactive 35S-radiolabelled probes. Biotinylated cRNA probes were labelled with either biotin-11-UTP or with allylamine-UTP, the latter method being able to produce a higher yield of labelled RNA. Different detection systems were tested, and hybridisation signal was seen in cells of anterior pituitary with both types of biotinylated probes. The signals were detected using either avidin-biotin-complex with peroxidase (ABC), peroxidase-anti-peroxidase (PAP) or gold-silver methods. ABC peroxidase detected using glucose oxidase-diaminobenzidine (DAB)-nickel solution appeared to be the best method for detecting labelled RNA probes, with very strong signal and low background. The biotinylated probes were comparable in sensitivity to the radiolabelled probes in detecting prolactin and GH mRNAs in the anterior lobe of the rat pituitary. These results indicate an alternative methods of labelling and detection of biotinylated probes which could have a potential role in research and diagnostic techniques.
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  • 7
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of calcitonin gene-related peptide (CGRP)-immunoreactive nerve fibres in the palate of rat, cat and monkey was studied using immunocytochemistry and radioimmunoassay. CGRP-containing nerve fibres were found, in all species studied, to form a rich plexus in the subepithelial and submucous layers, around excretory ducts and blood vessels. A small number of CGRP-containing nerve fibres penetrated the epithelium of the hard and soft palate, and terminated as free endings. Some CGRP-containing nerve fibres were found in the vicinity of the mucous glands. CGRP-immunoreactive motor end plates were seen in the striated muscle (tensor veli palatini) of the soft palate. Following capsaicin treatment a small depletion in CGRP-immunoreactive nerve fibres in the rat palate epithelium was noted. In contrast, CGRP immunoreactive fibres forming rich plexuses in other layers of the palate, including motor end plates, were not affected. The extractable CGRP showed no significant depletion (normal animals [n=10] 21.7±2.4 pmol/g compared with capsaicin-treated animals [n=10] 17.5±1.8 pmol CGRP/g wet weight). The reduction in the number of visible immunoreactive nerves following capsaicin application tends to confirm the sensory character of the CGRP-containing nerve fibres terminating in the epithelium of the hard and soft palate. The capsaicin insensitive CGRP-immunoreactive nerve fibres may thus have a predominantly motor function. The unilateral section of the mandibulary division of trigeminal, hypoglossal, glossopharyngeal and superior laryngeal nerve caused a depletion of variable severity in the ipsilateral CGRP-immunoreactive nerve fibres in the epithelium, subepithelial and periductal plexus and in motor end plates. These findings are in agreement with the radioimmunoassay results, which detect 22.7±1.0 pmol/g in normal tissues, 19.7±1.0 pmol/g after section of trigeminal nerve, and 18.6±1.2 pmol/g after simultaneous sectioning of the trigeminal, hypoglossal, glossopharyngeal, laryngeal and recurrent nerves. We conclude that CGRP is present in both sensory and motor components of cranial nerves which innervate the palate.
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  • 8
    ISSN: 1432-5233
    Keywords: Calcitonin gene-related peptide (CGRP) ; Substance P ; Diabetic BB rat ; Dorsal root ganglion (DRG) ; Spinal cord
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This study examined the experession of the sensory neuropeptides, calcitonin gene-related peptide (CGRP) and substance P (SP), in the lumbar 4 and 5 dorsal root ganglion (DRG) and spinal cord of spontaneously diabetic BB rats and non-diabetic controls using quantitative immunohistochemical analysis. In both animal groups immunoreactivities for CGRP and SP were widely distributed within the neurons of DRG and in nerve fibres of the dorsal spinal cord. Image analysis of each neuropeptide subpopulation in the DRG showed that in diabetic rats the cell diameter of immunostained CGRP neurons was significantly decreased compared with controls, while no difference could be found for SP-immunoreactive (IR) neurons. The decrease in the CGRP-IR cell diameter appeared to occur mainly in medium to large neurons (30–50 μm diameter; 2.2% controls, 〈1% diabetes), this change being parallel to an increased frequency of small-size neurons (〈20 μm diameter) in diabetic rats (62% controls, 69% diabetes;P〈0.05). However, there was no statistical difference in the total number of cells immunostained for either CGRP or SP between control and diabetic rats. The ratio of CGRP or SP neurons compared to total cells in the ganglion was similar in control and diabetic groups. No difference could be observed for peptide immunoreactivity in the dorsal and ventral horns of either control or diabetic animals. The observed changes of perikaryal size in diabetic rats might relate to the reduced axonal calibre and conduction velocity observed in these animals, and indicate that subpopulations of sensory neurons are affected differently by diabetes.
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  • 9
    ISSN: 1432-2307
    Keywords: Immunocytochemistry ; GFAP ; NSE ; Retinoblastoma ; S-100
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary An immunocytochemical study of 30 retinoblastomas was carried out using antibodies to neuronal and glial markers. The tumours were found to react with antibodies to neuron-specific enolase (NSE), a marker for neuronal elements, and S-100 and glial fibrillary acidic protein (GFAP), both of which are proteins present in glia. Two distinct cell populations were found within the tumour: the first, composed of anaplastic tumour cells at various stages of differentiation, showed both NSE and S-100 immunoreactivity; the second cell type, which immunostained for S-100 and GFAP, resembled mature glial cells. The results of this study indicate that the retinoblastoma may arise from a pluripotential primitive cell partially retaining neuronal and glial characteristics.
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  • 10
    ISSN: 1432-0428
    Keywords: VIP ; radioimmunoassay ; immunocytochemistry ; eyes ; urinary bladder ; prediabetes ; diabetic Chinese hamsters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The eyes and urinary bladder of non-diabetic, prediabetic and diabetic Chinese hamsters were evaluated by radioimmunoassay and immunocytochemistry to determine the content and distribution of vasoactive intestinal peptide (VIP). The average concentration of VIP was increased in the eyes of all diabetic (pmol/g = 68%, pmol/organ = 50%) and prediabetic (pmol/g = 152%, pmol/organ = 115%) hamsters compared with age-matched non-diabetic animals. Immunocytochemistry showed that the elevation of VIP was primarily related to greater intensity of fluorescence of the nerve fibres in the vasculature of the choroid. The average content of VIP in the urinary bladder was greater in diabetic animals only on the basis of pmol/organ (135%) and in prediabetics on the basis of pmol/g (87%) compared with non-diabetic animals. Qualitative immunocytochemistry suggested that the elevated level of VIP was related to a larger distribution of nerve fibres in the urinary bladder of diabetic hamsters. The high level of VIP in the eyes and urinary bladder of diabetic and prediabetic hamsters is an interesting observation which should receive further study to determine whether it is an aetiological agent underlying the pathogenesis of ophthalmic complications and neurogenic bladder or the result of some pathological process which affects these organs.
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