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In vitro Autoradiographic Localization of Amino Acid Receptors and Uptake Sites

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Amino Acids

Part of the book series: Neuromethods ((NM,volume 3))

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Abstract

Some amino acids may function as neurotransmitters and are presumably released at nerve terminals to interact with specific receptors postsynaptically. Biochemical studies of high-affinity uptake, a potential marker of nerve terminals and of receptor binding, have been employed successfully to elucidate the role of amino acids in the central nervous system (W Walz and J.W. Ferkany, this volume). The recent use of autoradiographic techniques to study uptake and receptors has permitted much greater anatomical resolution than homogenate methods. Other advantages include the ability to study different conditions in consecutive sections, to study human tissues, and to circumvent the blood-brain barrier.

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References

  • Alexander G. M., Schwartzman R J, Bell R D, Yu J., and Renthal A. (1981) Quantitative measurement of local cerebral metabolic rate for glucose utilizing tritiated 2-deoxyglucose. Brain Res 223, 59–67

    Article  PubMed  CAS  Google Scholar 

  • Berger M. and Ben-Ari Y. (1983) Autoradiographic visualization of [3H] kainic acid receptor subtypes in the rat hippocampus. Neurosci. Lett 39, 237–242.

    Article  PubMed  CAS  Google Scholar 

  • Biegon A. and Rainbow T C. (1983) Localization and characterization of [3H] desmethylimipramine binding sites in rat brain by quantitative autoradrography J Neurosci 5, 1069–1076.

    Google Scholar 

  • Biziere K., Thompson H., and Coyle J T (1980) Characterization of specific, high-affinity binding sites for L-[3H]-glutamic acid in rat brain membranes. Brain Res 183, 421–433

    Article  PubMed  CAS  Google Scholar 

  • Cuatrecasas P. and Hollenberg M. D. (1976) Membrane receptors and hormone action. Adv. Protein Chem. 30, 251–451

    Article  PubMed  CAS  Google Scholar 

  • DeBarry J., Langley O. K., Vincendon G, and Gombos G (1982) L-Glutamate and L-glutamine uptake in adult rat cerebellum. an autoradrographic study Neuroscience 7, 1289–1297.

    Article  CAS  Google Scholar 

  • Ehinger B. (1981) [3H]-D-Aspartate accumulation in the retina of pigeon, guinea pig and rabbit Exp Eye Res. 33, 381–391

    Article  PubMed  CAS  Google Scholar 

  • Foster A. C., Mena E. E, Monaghan D T., and Cotman C. W (1981) Synaptic localization of kainic acid binding sites Nature 289, 73–75

    Article  PubMed  CAS  Google Scholar 

  • Geary W A, II, and Wooten G. F (1983) Quantitative film autoradiography of opiate agonist and antagonist binding in rat brain. J. Pharmacol Exp. Ther. 225, 234–240

    PubMed  CAS  Google Scholar 

  • Greenamyre J. T, Young A. B, and Penney J B. (1984) Quantitative autoradiographic distribution of L-[3H]-glutamate-binding sites in rat central nervous system. J Neurosci. 4, 2133–2144.

    PubMed  CAS  Google Scholar 

  • Greenamyre J T., Young A B, and Penney J B. (1983) Quantitative autoradiography of L-[3H] glutamate binding to rat brain. Neurosci. Lett 37, 155–160

    Article  PubMed  CAS  Google Scholar 

  • Halpain S., Parsons B, and Rainbow T C. (1983) Tritium-film autoradiography of sodium-independent glutamate binding sites in rat brain Eur J. Pharmacol 86, 313–314.

    Article  Google Scholar 

  • Hampton C K and Redburn D. A (1983) Autoradiographic analysis of 3H-glutamate, 3H-dopamine and 3H-GABA accumulation in rabbit retma after kainic acid treatment. J. Neurosci. Res 9, 239–251

    Article  PubMed  CAS  Google Scholar 

  • Hattori T, McGeer P L, Fibiger H. C, and McGeer E. G (1973) On the source of GABA-containing terminals in the substantia nigra Electron microscopic autoradiographic and biochemical studies Brain Res 54, 103–114

    Article  PubMed  CAS  Google Scholar 

  • Henke H, Beaudet A, and Cuenod M (1981) Autoradiographic localization of specific kainic acid binding sites in pigeon and rat cerebellum Brain Res 219, 95–105

    Article  PubMed  CAS  Google Scholar 

  • Herkenham M, and Pert C. B (1982) Light microscopic localization of brain opiate receptors a general autoradiographic method which preserves tissue quality J Neurosci 2, 1129–1149.

    PubMed  CAS  Google Scholar 

  • Hokfelt T and Ljungdahl Å. (1970) Cellular localization of labeled gamma-ammobutyric acid (H-GABA) in rat cerebellar cortex. an autoradiographic study. Brain Res. 22, 391–396.

    Article  PubMed  CAS  Google Scholar 

  • Hokfelt T and Ljungdahl Å (1971) Light and electron microscopic autoradiography on spinal cord slices after incubation with labeled glycine. Brain Res 32, 189–194

    Article  PubMed  CAS  Google Scholar 

  • Iversen L. L and Bloom F E. (1972) Studies of the uptake of 3H-GABA and 3H-glycine in slices and homogenates of rat brain and spinal cord by election microscopic autoradiography Brain Res 41, 131–143

    Article  PubMed  CAS  Google Scholar 

  • Kuhar M J. (1982) Localization of drug and neurotransmitter receptors in brain by light microscopic autoradiography, in Handbook of Psychopharmacology (Iversen L L, Iversen S.D., and Snyder S.H, eds.), pp. 299–320 Plenum Press, New York.

    Google Scholar 

  • Kuhar M J (1983) Autoradiographic localization of drug and neurotransmitter receptors, in Handbook of Chemical Neuroanatomy Vol 1. Methods of Chemical Neuroanatomy (Bjorklund A and Hokfelt T, eds.), pp 398–415 Elsevier, New York

    Google Scholar 

  • Kuhar M J (1984) Receptor localization with the microscope, in Neurotransmitter Receptor Binding (Yamamura H.I, Enna S J, and Kuhar M J, eds), in press, Raven Press, New York.

    Google Scholar 

  • Kuhar M J and Unnerstall J R (1982) In vitro labeling receptor autoradiography loss of label during ethanol dehydration and preparative procedures Brain Res 244, 178–181

    Article  PubMed  CAS  Google Scholar 

  • Lysz T., Toga A W, and Geary W A, II (1982) Standardization of 3H-tissue images with different lsotopic standards Soc Neurosci Abstract 8, 645.

    Google Scholar 

  • Marshall J and Voaden M (1975) Autoradiographic identification of the cells accumulating 3H-γ-ammobutyric acid in mammalian retinae a species comparison. Vision Res 15, 459–461.

    Article  PubMed  CAS  Google Scholar 

  • Matus A. I. and Dennison M E. (1971) Autoradiographic localization of tritiated glycine at “flat-vesicle” synapses in spinal cord Brain Res 32, 195–197

    Article  PubMed  CAS  Google Scholar 

  • Mohler H, Richards J. G, and Wu J-Y (1981) Autoradiographic localization of benzodiazepine receptors in immunocytochemically identified γ-aminobutyric synapses. Proc. Natl Acad Sci USA 78, 1935–1938.

    Article  PubMed  CAS  Google Scholar 

  • Monaghan D T and Cotman C W (1982) The distribution of [3H] kainic acid binding sites in rat CNS as determined by autoradiography. Brain Res 252, 91–100

    Article  PubMed  CAS  Google Scholar 

  • Monaghan D. T, Holets V R., Toy D W, and Cotman C W (1983a) Anatomical distributions of four pharmacologically distinct 3H-L-glutamate binding sites. Nature 306, 176–179.

    Article  PubMed  CAS  Google Scholar 

  • Monaghan D T., McMills M C., Chamberlin A R, and Cotman C W (1983b) Synthesis of [3H]2-amino-4-phosphonobutyric acid and characterization of its binding to rat brain membranes. a selective ligand for the chloride/calcium-dependent class of L-glutamate binding sites Brain Res 278, 137–144.

    Article  PubMed  CAS  Google Scholar 

  • Palaclos J. M. and Kuhar M. J (1982) Ontology of high-affinity GABA and benzodiazepine receptors in rat cerebellum an autoradiographic study. Brain Res 2, 531–539

    Google Scholar 

  • Palacios J M, Young W S., III, and Kuhar M J (1980) Autoradiographic localization of GABA receptors in the rat cerebellum Proc Nutl Acad Sci USA 77, 670–674.

    Article  CAS  Google Scholar 

  • Palaclos J M., Niehoff D L, and Kuhar M. J (1981a) Receptor autoradiography with tritium-sensitive film: potential for computerized densitometry. Neurosci Lett 25, 101–105

    Article  Google Scholar 

  • Palacios J M., Unnerstall J.R, Young W S., III, and Kuhar M. J (1981b) Radiohistochemical studies of benzodiazepine and GABA receptors and their interactions, in GABA and Benzodiazepine Receptors (Costa E., Di Chiara G, and Gessa G L, eds.), pp. 53–60 Raven Press, New York

    Google Scholar 

  • Palacios J M, Wamsley J K. and Kuhar M J (1981c) GABA, benzodiazepine, and histamine-H1 receptors in the guinea pig cerebellum effects of kainic acid injections studied by autoradiographic methods. Brain Res 214, 155–162

    Article  PubMed  CAS  Google Scholar 

  • Palacios J M, Wamsley J K, and Kuhar M J (1981d) High affinity GABA receptors-autoradiographic localization. Brain Res 222, 285–307

    Article  PubMed  CAS  Google Scholar 

  • Pan H S., Frey K. A, Young A B., and Penney J B, Jr (1983) Changes in [3H] muscimol binding in substantia nigra, entopeduncular nucleus, globus pallidus, and thalamus after striatal lesions as demonstrated by quantitative receptor autoradiography. J. Neurosci 3, 1189–1198

    PubMed  CAS  Google Scholar 

  • Parsons B. and Rainbow T C (1983) Quantitative autoradiography of sodium-dependent [3H]D-aspartate binding sates in rat brain Neurosci Lett 36, 9–12

    Article  PubMed  CAS  Google Scholar 

  • Penney J B, Jr., Pan H S, Young A. B., Frey K. A., and Dauth G. W. (1981) Quantitative autoradiography of [3H] muscimol binding in rat brain Science 214, 1036–1038.

    Article  PubMed  CAS  Google Scholar 

  • Penney J. B. and Young A B (1982) Quantitative autoradiography of neurotransmitter receptors in Huntington disease. Neurology 32, 1391–1395.

    PubMed  Google Scholar 

  • Rainbow T C, Biegon A., and McEwen B. S. (1982a) Autoradiographic localization of imipramine binding in rat brain Eur J. Pharmacol 77, 363–364

    Article  PubMed  CAS  Google Scholar 

  • Rainbow T C, Bleisch W. V., Biegon A., and McEwen B. S (1982b) Quantitative densitometry of neurotransmitter receptors. J Neurosci Meth 5, 127–138

    Article  CAS  Google Scholar 

  • Rogers A W. (1979) Techniques of Autoradiography. Elsevier, New York

    Google Scholar 

  • Roth L.J., Diab I M, Watanabe M., and Dinerstein R. (1974) A correlative radioautographic fluorescent and histochemical technique for cytopharmacology. Mel Pharmacol. 10, 986–998.

    CAS  Google Scholar 

  • Sarthy P. V. (1983) Release of [3H]γ-aminobutyric acid from glial (Muller) cells of the rat retina effects of K+, veratridine, and ethylenediamine J Neurosci 12, 2494–2503.

    Google Scholar 

  • Schulze E. and Neuhoff V. (1983) Uptake, autoradiography and release of taurine and homotaurine from retinal tissue Int. J. Neurosci 18, 253–268

    Article  PubMed  CAS  Google Scholar 

  • Taylor, N R., Unnerstall, J R, Mashal, R. D, DeSouza, E. B., and Kuhar, M. J. (1984) Receptor autoradiography: coping with regional differences in autoradiographic efficiency with tritium Soc. Neurosci Abs. 10, 557

    Google Scholar 

  • Unnerstall J. R. and Wamsley J. K. (1983) Autoradiographic localization of high-affinity [3H]-kainic acid binding sites in the rat forebrain Eur J, Pharmacol 86, 361–371.

    Article  CAS  Google Scholar 

  • Unnerstall J R., Kuhar M. J., Niehoff D L. and Palacios J. M. (1981) Benzodiazepine receptors are coupled to a subpopulation of γ-aminobutyric acid (GABA) receptors evidence from a quantitative autoradiographic study. J Pharmacol. Exp. Ther. 218, 797–804.

    PubMed  CAS  Google Scholar 

  • Unnerstall J R, Niehoff D. L, Kuhar M. J. and Palacios J M. (1982) Quantitative receptor autoradiography using [3H]Ultrofilm application to multiple benzodiazepine receptors. J. Neurosci Meth. 6, 59–73

    Article  CAS  Google Scholar 

  • Wamsley J. K. and Palacios J M (1983) Autoradiographic distribution of receptor ligand binding-amino acid and benzodiazepine receptors. Handbook of Chemical Neuroanatomy Vol. 2-Classical Transmitters in the CNS. (Bjorklund A, Hokfelt T, and Kuhar M.J, eds), in press, Elsevier, New York

    Google Scholar 

  • Whitehouse P J, Wamsley J K, Zarbin M A, Price D L, Tourtellotte W.W., and Kuhar M.J (1983) Amyotrophic lateral sclerosis. alterations in neurotransmitter receptors Ann Neural 14, 8–16

    Article  CAS  Google Scholar 

  • Wilkin G. P, Csillag A, Balázs R., Kingsbury A E, Wilson J E, and Johnson A. L (1981) Localization of high affinity [3H] glycine transport sites in the cerebellar cortex Brain Res 216, 11–33

    Article  PubMed  CAS  Google Scholar 

  • Wilkin G P., Hudson A L, Hill D R., and Bowery N. G (1981) Autoradiographic localization of GABAB receptors in rat cerebellum Nature 294, 584–587

    Article  PubMed  CAS  Google Scholar 

  • Yamamura H. I., Enna S J. and Kuhar M. J, eds. (1984) Neurotransmitter Receptor Binding. In press, Raven Press, New York

    Google Scholar 

  • Yazulla S. (1981) GABAergic synapses in the goldfish retina: an autoradiographic study of 3H-musciomol and 3H-GABA binding. J Comp. Neurl 200, 83–93

    Article  CAS  Google Scholar 

  • Young A. B. and Snyder S H (1973) Strychnine binding associated with glycine receptors of the central nervous system. Proc Natl Acad Sci. USA 70, 2832–2836

    Article  PubMed  CAS  Google Scholar 

  • Young W. S, III and Kuhar M J. (1979a) A new method for receptor autoradiography [3H] opioid receptors in rat brain Brain Res 179, 255–270.

    Article  PubMed  CAS  Google Scholar 

  • Young W. S, III and Kuhar M. J. (1979b) Autoradiographic localization of benzodiazepine receptors in the brains of humans and animals Nature 280, 393–395

    Article  Google Scholar 

  • Young W S, III and Kuhar M. J (1980) Radiohistochemical localization of benzodiazepine receptors in rat brain J Phrmacol Exp Ther 212, 337–346

    CAS  Google Scholar 

  • Young W S., III and Kuhar M J. (1981) The light microscopic radiohistochemistry of drugs and neurotransmitter receptors using diffusible ligands, in Current Trends in Morphological Techniques, Vol III (Johnson J E., Jr, Adelman R.C. and Roth G S, eds), pp 119–135 CRC Press. Boca Raton, FL

    Google Scholar 

  • Zaczek R, Koller K., Cotter R, Heller D., and Coyle J T (1983) N-Acetylaspartylglutamate. an endogenous peptide with high affinity for a brain. Proc. Natl Acad. Sci USA 80, 1116–1119.

    Article  PubMed  CAS  Google Scholar 

  • Zarbin M. A, Wamsley J. K., and Kuhar M J (1981) Glycine receptor light microscopic autoradiographic localization with [3H] strychnine. J. Neurosci 1, 532–547.

    PubMed  CAS  Google Scholar 

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Alan A. Boulton Glen B. Baker James D. Wood

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Scott Young, W. (1985). In vitro Autoradiographic Localization of Amino Acid Receptors and Uptake Sites. In: Boulton, A.A., Baker, G.B., Wood, J.D. (eds) Amino Acids. Neuromethods, vol 3. Humana Press. https://doi.org/10.1385/0-89603-077-6:179

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  • DOI: https://doi.org/10.1385/0-89603-077-6:179

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-077-0

  • Online ISBN: 978-1-59259-608-9

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