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  • 1
    ISSN: 1573-6830
    Keywords: methionine-enkephalin ; angiotensin-converting enzyme ; tyrosine hydroxylase ; vasopressin ; computerized microdensitometry ; quantitative autoradiography ; quantitative immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. We have used horseradish peroxidase-conjugated protein A- and125I-protein A to develop immunohistochemical and radioimmunohistochemical methods for the localization of antigens in brain and other tissues of the rat. 2. We visualized methionine-enkephalin fibers in the rat brain by incubating tissue sections with a specific polyclonal antibody and peroxidase-conjugated protein A. The method is simple, fast, and less expensive and more sensitive than classical immunohistochemical techniques and the principle could be used to visualize many other tissue antigens. 3. Incubation of tissue samples with specific polyclonal antibodies and125I-protein A, followed by autoradiography, allows the permanent recording of the radiommunohistochemical localization of brain methionine-enkephalin, tyrosine hydroxylase, and angiotensin-converting enzyme and of pituitary vasopressin and could be applied to the localization of many other tissue antigens. 4. A new quantitative radioimmunohistochemical technique for methionine-enkephalin allows the determination of the endogenous peptide content in discrete brain nuclei from 16-µm-thick sections. The method is based on the quantitative determination of the amount of125I-protein A bound to specific tissue areas after incubation with a specific polyclonal antibody, followed by autoradiography and computerized microdensitometry. To quantify the endogenous peptide content, the values obtained are interpolated into a methionine-enkephalin internal standard curve. This standard curve was constructed by measuring endogenous concentrations of methionine-enkephalin by radioimmunoassay in specific brain regions and correlating these values with quantitative autoradiographic determinations in homologous areas of adjacent sections. Similar methods can be developed for other tissue antigens. 5. These new methods allow for the localization and quantification of tissue antigens in very discrete areas of the brain and other tissues and have a wide application in neurobiology and pathology.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Cellular and molecular neurobiology 9 (1989), S. 273-279 
    ISSN: 1573-6830
    Keywords: insulin ; insulin receptors ; neuropeptide receptors ; invertebrate nervous system ; invertebrate ganglia ; quantitative autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. Insulin binding sites were characterized and quantified in snail (Helix aspersa) ganglia by incubation of tissue sections with125I-porcine insulin, autoradiography with [3H]Ultrofilm, image analysis coupled to computer-assisted microdensitometry, and comparison with125I-standards. Cellular localization was performed in the same sections by emulsion autoradiography. 2. Specific insulin binding sites were demonstrated in discretely localized groups of neurons of the cerebral, pleural, parietal, visceral, and pedal ganglia and in nerves. Scatchard analysis performed with consecutive sections from single animals revealed a single class of high-affinity insulin binding sites (K d, 0.13 ± 0.01 nM;B max, 157 ± 10 fmol/mg protein). 3. Our results suggest that insulin may play a role as a neurotransmitter or neuromodulator in snail ganglia.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Cellular and molecular neurobiology 7 (1987), S. 151-173 
    ISSN: 1573-6830
    Keywords: neuropeptides ; atrial natriuretic factor ; atriopeptins ; quantitative autoradiography ; receptors ; circumventricular organs ; dehydration ; hypertension ; cardiovascular control ; fluid metabolism ; choroid plexus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. We have studied the localization, kinetics, and regulation of receptors for the circulating form of the atrial natriuretic peptide (ANP; 99–126) in the rat brain. 2. Quantitative autoradiographic techniques and a125I-labeled ligand,125I-ANP (99–126), were employed. Afterin vitro autoradiography, quantification was achieved by computerized microdensitometry followed by comparison with125I-standards. 3. ANP receptors were discretely localized in the rat brain, with the highest concentrations in circumventricular organs, the choroid plexus, and selected hypothalamic nuclei involved in the production of the antidiuretic hormone vasopressin and in blood-pressure control. 4. Spontaneously (genetic) hypertensive rats showed much lower numbers of ANP receptors than normotensive controls in the subfornical organ, the area postrema, the nucleus of the solitary tract, and the choroid plexus. These changes are in contrast to those observed for receptors of angiotensin II, another circulating peptide with actions opposite to those of ANP. 5. Under conditions of acute dehydration after water deprivation, as well as under conditions of chronic dehydration such as those present in homozygous Brattleboro rats, there was an up-regulation of ANP receptors in the subfornical organ. 6. Our results indicate that in the brain, circumventricular organs contain ANP receptors which could respond to variations in the concentration of circulating ANP. In addition, brain areas inside the blood-brain barrier contain ANP receptors probably related to the endogenous, central ANP system. 7. The localization of ANP receptors and the alterations in their regulation present in genetically hypertensive rats and after dehydration indicate that brain ANP receptors are probably related to fluid regulation, including the secretion of vasopressin, and to cardiovascular function. ANP and angiotensin II could act as mutual antagonists in the brain as they do in the periphery. 8. ANP receptors in the choroid plexus may be related to the formation of cerebrospinal fluid.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-6830
    Keywords: quantitative autoradiography ; receptor autoradiography ; neuropeptide receptors ; angiotensin II receptors ; receptor quantitation in brain nuclei
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. Autoradiographic techniques coupled with computerized microdensitometry and comparison with125I standards were used to characterize and quantitate receptors for neuropeptides in rat brain and adrenal and pituitary glands. 2. These techniques are rapidly performed, anatomically precise, and more sensitive than membrane binding techniques. They permit the determination of complete saturation curves and Scatchard analysis in discrete nuclei of the rat brain and in single rat pituitary and adrenal glands. 3. Angiotensin II (AII) receptors were quantitated after incubation of 16-µm tissue sections with the AII agonist125I-[Sar1]-AII. 4. High-affinity, high-density AII receptors were present in the organon subfornicalis, organon vasculosum laminae terminalis and nuclei triangularis septalis, suprachiasmatis, and paraventricularis of the rat and in rat adrenal capsule-zona glomerulosa area, adrenal medulla, and anterior pituitary. 5. These techniques could be used for precise localization and quantitation of other neuropeptide receptors in single rat brain nuclei, after optimizing the assay conditions and provided that suitable125I ligands are available.
    Type of Medium: Electronic Resource
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