Autoradiographic Localization of Brain Peptide Receptors at the Electron Microscopic Level

  • A. Beaudet
  • E. Hamel
  • K. Leonard
  • M. Vial
  • E. Moyse
  • P. Kitabgi
  • J. P. Vincent
  • W. Rostène

Abstract

Neurotransmitters and related drugs exert their biochemical, electrophysiological, and, ultimately, behavioral effects by acting upon specific receptor molecules embedded in neuronal and, in some cases, perhaps also glial plasma membranes. This interaction involves: (1) recognition of a specific binding site on the surface of the receptor and (2) translation of the recognition information into a response signal. Pioneering studies by Clark (1933) demonstrated that the binding of drugs (ligands) to receptors was reversible, obeyed the law of mass action, and occurred at very low concentrations of ligand. Considerable progress in our understanding of receptor mechanisms was later to emerge from the development of sensitive methods for measuring the receptor-specific binding of radiolabeled ligand probes (Snyder and Bennett, 1976). Not only did the availability of such methods greatly facilitate the biophysical, pharmacological, and molecular characterization of receptors for neurotransmitters, but it also made possible their localization in situ, using autoradiographic techniques.

Keywords

Dopamine Iodine Morphine Serotonin Neurol 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Altschuler, R. A., Betz, H., Parakkal, M. H., Reeks, K. A., and Wenthold, R. J., 1986, Identification of glycinergic synapses in the cochlear nucleus through immunocytochemical localization of the postsynaptic receptor, Brain Res. 369:316–320.PubMedCrossRefGoogle Scholar
  2. Antkiewicz-Michaluk, L., Havemann, U., Vetulani, J., Wellstein, A, and Kuschinsky, K., 1984, Opioid-specific recognition sites of the mu- and the delta-type in rat striatum after lesions with kainic acid, Life Sci. 35:347–355.PubMedCrossRefGoogle Scholar
  3. Arimatsu, Y., Seto, A., and Amano, T., 1978, Localization of α-bungarotoxin binding sites in mouse brain by light and electron microscopic autoradiography, Brain Res. 147:165–169.PubMedCrossRefGoogle Scholar
  4. Atweh, S. F., and Kuhar, M. J., 1977, Autoradiographic localization of opiate receptors in rat brain. III. The telencephalon, Brain Res. 134:393–405.PubMedCrossRefGoogle Scholar
  5. Beaudet, A., and Descarries, L., 1978, The monoamine innervation of rat cerebral cortex: Synaptic and nonsynaptic axon terminals, Neuroscience 3:851–860.PubMedCrossRefGoogle Scholar
  6. Beaudet, A., Trémeau, O., Ménez, A., and Droz, B., 1979, Visualisation des récepteurs aux opiacés dans le locus coeruleus du rat: Étude radioautographique à haute resolution après administration d’un analogue tritié de la met-enképhaline, CR. Acad. Sci. 289:591–594.Google Scholar
  7. Beaudet, A., Leonard, K., Vial, M., Moyse, E., Kitabgi, P., Vincent, J. P., and Rostène W., 1987, Electron microscopic localization of neurotensin receptors in the substantia nigra of the rat, Soc. Neurosci. Abstr. 13:563.Google Scholar
  8. Blackett, N. M., and Parry, D. M., 1977, A simplified method of “hypothetical grain” analysis of electron microscope autoradiographs, J. Histochem. Cytochem. 25:206–214.PubMedCrossRefGoogle Scholar
  9. Bowen, W. D., Pert, C. B., and Pert, A., 1982, Nigral 6-hydroxydopamine lesions equally decrease μ and δ opiate binding to striatal patches: Further evidence for a conformationally malleable type 1 opiate receptor, Life Sci. 31:1679–1682.PubMedCrossRefGoogle Scholar
  10. Chang, K. J., Cooper, B. R., Hazum, E., and Cuatrecasas, P., 1979, Multiple opiate receptors: Different regional distribution in the brain and differential binding of opiates and opioid peptides, Mol. Pharmacol. 16:91–104.PubMedGoogle Scholar
  11. Childers, S. R., Schwarcz, R., Coyle, J. T., and Snyder, S. H., 1978, Radioimmunoassay of enkephalins: Levels of methionine- and leucine-enkephalin in morphine dependent and kainic acid lesioned rat brains, in: Advances in Biochemical Psychopharmacology, Vol. 18 (E. Costa and M. Trabucchi, eds.), Raven Press, New York, pp. 161–173.Google Scholar
  12. Clark, A. J., 1933, The Mode of Action of Drugs on Cells, Arnold, London.Google Scholar
  13. Cuello, A. C., 1983, Nonclassical neuronal communication, Fed. Proc. Fed. Am. Soc. Exp. Biol. 42:2912–2922.Google Scholar
  14. Dana, C, Vial, M., Kitabgi, P., Leonard, K., Beaudet, A., and Rostène, W., 1985, Distribution radioautographique et ultrastructurale des sites de liaison de la neurotensine dans le cerveau de rat, Ann. Endocrinol. 46:16N.Google Scholar
  15. Dana, C, Leonard, K., Vial, M., Rostène, W., and Beaudet, A., 1987, Autoradiographic localization of neurotensin binding sites in rat ventral tegmental area at the electron microscopic level, Neuroscience 22:5785.Google Scholar
  16. Demoliou-Mason, C. D., and Barnard, E. A., 1986, Distinct subtypes of the opioid receptor with allosteric interactions in brain membranes, J. Neurochem. 46:1118–1128.PubMedCrossRefGoogle Scholar
  17. Descarries, L., and Beaudet, A., 1983, The use of radioautography for investigating transmitter-specific neurons, in: Handbook of Chemical Neuroanatomy, Vol. 1 (A. Björklund and T. Hökfelt, eds.), Elsevier, Amsterdam, pp. 286–364.Google Scholar
  18. Descarries, L., Beaudet, A., and Watkins, K. C., 1975, Serotonin nerve terminals in adult rat neocortex, Brain Res. 100:563–588.PubMedCrossRefGoogle Scholar
  19. Gardner, E. L., Zukin, S. R., and Makman, M. H., 1980, Modulation of opiate receptor binding in striatum and amygdala by selective mesencephalic lesions, Brain Res. 194:232–239.PubMedCrossRefGoogle Scholar
  20. Goodman, R. R., Snyder, S. H., Kuhar, M. J., and Young, W. S., III, 1980, Differentiation of delta and mu opiate receptor localizations by light microscopic autoradiography, Proc. Natl. Acad. Sci. USA 77:6239–6243.PubMedCrossRefGoogle Scholar
  21. Hamel, E., and Beaudet, A., 1984a, Localization of opioid binding sites in rat brain by electron microscopic radioautography, J. Electron Microsc. Tech. 1:317–329.CrossRefGoogle Scholar
  22. Hamel, E., and Beaudet, A., 1984b, Electron microscopic autoradiographic localization of opioid receptors in rat neostriatum, Nature 312:155–157.PubMedCrossRefGoogle Scholar
  23. Hamel, E., and Beaudet, A., 1987, Opioid receptors in rat neostriatum: Radioautographic distribution at the electron microscopic level, Brain Res. 401:239–257.PubMedCrossRefGoogle Scholar
  24. Hazum, E., Chang, K.-J., and Cuatrecasas, P., 1980, Cluster formation of opiate (enkephalin) receptors in neuroblastoma cells: Differences between agonists and antagonists and possible relationships to biological functions, Proc. Natl. Acad. Sci. USA 77:3038–3041.PubMedCrossRefGoogle Scholar
  25. Herkenham, M., 1984, Autoradiographic demonstration of receptor distributions, in: Brain Receptor Methodologies, Part A (P. J. Marangos, I. C. Campbell, and R. M. Cohen, eds.), Academic Press, New York, pp. 127–152.Google Scholar
  26. Herkenham, M., and McLean, S., 1986, Mismatches between receptor and transmitter localizations in the brain, in: Quantitative Receptor Autoradiography, Vol. 19 (C. A. Boast, E. A. Snowhill, and C. A. Altar, eds.), Liss, New York, pp. 137–171.Google Scholar
  27. Herkenham, M., and Pert, C.B., 1982, Light microscopic localization of brain opiate receptors: A general auto-radiographic method which preserves tissue quality, J. Neurosci. 2:1129–1149.PubMedGoogle Scholar
  28. Hökfelt, T., Everitt, B. J., Theodorsson-Norheim, E., and Goldstein, M., 1984, Occurrence of neurotensinlike immunoreactivity in subpopulations of hypothalamic, mesencephalic, and medullary catecholamine neurons, J. Comp. Neurol. 222:543–559.PubMedCrossRefGoogle Scholar
  29. Hollt, V., and Herz, A., 1978, In vivo receptor occupation by opiates and correlation to the pharmacological effect, Fed. Proc. Fed. Am. Soc. Exp. Biol. 37:158–161.Google Scholar
  30. Hunt, S. P., and Schmidt, J., 1978, The electron microscopic autoradiographic localization of α-bungarotoxin binding sites within the central nervous system of the rat, Brain Res. 142:152–159.PubMedCrossRefGoogle Scholar
  31. Jacob, M. H., and Berg, D. K., 1983, The ultrastructural localization of α-bungarotoxin binding sites in relation to synapses on chick ciliary ganglion neurons, J. Neurosci. 3:260–271.PubMedGoogle Scholar
  32. Jan, Y. N., and Jan, L. Y., 1983, A LHRH-like peptidergic neurotransmitter capable of ‘action at a distance’ in autonomic ganglia, Trends Neurosci. 6:320–325.CrossRefGoogle Scholar
  33. Kessler, J. P., Kitabgi, P., and Beaudet, A., 1986, High affinity neurotensin binding to afferent and efferent components of the vagal complex, Soc. Neurosci. Abstr. 12:811.Google Scholar
  34. Kitabgi, P., Poustis, C, Granier, C, Van Rietschoten, J., Rivier, J., Morgat, J. L., and Freychet, P., 1980, Neurotensin binding to extraneuronal and neural receptors: Comparison with biological activity and structure activity relationships, Mol. Pharmacol. 18:11–19.PubMedGoogle Scholar
  35. Kitabgi, P., Checler, F., Mazella, J., and Vincent, J. P., 1985, Pharmacology and biochemistry of neurotensin receptors, Rev. Clin. Basic Pharmacol. 5:397–486.Google Scholar
  36. Kuhar, M. J., and Yamamura, H. I., 1974, Light microscopic autoradiographic localization of cholinergic muscarinic sites in rat brain, Proc. Soc. Neurosci. 4:29.Google Scholar
  37. Kuhar, M. J., Taylor, M., Wamsley, J. K., Hulme, E. C., and Birdsall, N. J. M., 1981, Muscarinic cholinergic receptor localization in brain by electron microscopic autoradiography, Brain Res. 216:1–9.PubMedCrossRefGoogle Scholar
  38. Lentz, T. L., and Chester, J., 1977, Localization of acetylcholine receptors in central synapses, J. Cell Biol. 75:258–267.PubMedCrossRefGoogle Scholar
  39. Lewis, M. E., Khachaturian, H., and Watson, S. J., 1985, Combined autoradiographic-immunocytochemical analysis of opioid receptors and opioid peptide neuronal systems in brain, Peptides 6:37–47.PubMedCrossRefGoogle Scholar
  40. McLean, S., Rothman, R. B., and Herkenham, M., 1986, Autoradiographic localization of μ,- and δ-opiate receptors in the forebrain of the rat, Brain Res. 378:49–60.PubMedCrossRefGoogle Scholar
  41. Miller, M. M., Billiar, R. B., and Beaudet, A., 1987, Ultrastructural distribution of alpha-bungarotoxin binding sites in the suprachiasmatic nucleus of the rat hypothalamus, Cell Tissue Res. (in press).Google Scholar
  42. Miller, R. J., Chang, K.-J., Leighton, J., and Cuatrecasas, P., 1978, Interaction of iodinated enkephalin analogs with opiate receptors, Life Sci. 22:379–388.PubMedCrossRefGoogle Scholar
  43. Mohler, H., Richards, J. G., and Wu, J.-Y., 1981, Autoradiographic localization of benzodiazepine receptors in immunocytochemically identified γ-aminobutyrergic synapses, Proc. Natl. Acad. Sci. USA 78:1935–1938.PubMedCrossRefGoogle Scholar
  44. Moyse, E., Vial, M., Leonard, K., Kitabgi, P., Vincent, J. P., Rostène, W., and Beaudet, A., 1985, Electron microscopic visualization of neurotensin binding sites in rat substantia nigra, Soc. Neurosci. Abstr. 11:415.Google Scholar
  45. Moyse, E., Pasquini, F., Quirion, R., and Beaudet, A., 1986, 125I-FK 33–824: A selective probe for radio-autographic labeling of μ opioid receptors in the brain, Peptides 7:351–355.PubMedCrossRefGoogle Scholar
  46. Moyse, E., Rostène, W., Vial, M., Leonard, K., Mazella, J., Kitabgi, P., Vincent, J. P., and Beaudet, A., 1987, Regional distribution of neurotensin binding sites in rat brain: A light microscopic radioautographic study usingmonoiodo 125I-Tyr3-neurotensin, Neuroscience 22:525–536. PubMedCrossRefGoogle Scholar
  47. Murrin, L. C., Coyle, J. T., and Kuhar, M. J., 1980, Striatal opiate receptors: Pre- and post-synaptic localization, Life Sci. 27:1175–1183.PubMedCrossRefGoogle Scholar
  48. Palacios, J. M., and Kuhar, M. J., 1981, Neurotensin receptors are located on dopamine-containing neurons in rat midbrain, Nature 294:587–589.PubMedCrossRefGoogle Scholar
  49. Palacios, J. M., Niehoff, D. L., and Kuhar, M. J., 1981, Receptor autoradiography with tritium-sensitive film: Potential for computerized densitometry, Neurosci. Lett. 25:101–105.PubMedCrossRefGoogle Scholar
  50. Parenti, M., Titrone, F., Olgiati, V. R., and Gropetti, A., 1983, Presence of opiate receptors on striatal serotoninergic nerve terminals, Brain Res. 280:317–322.PubMedCrossRefGoogle Scholar
  51. Pert, C. B., Kuhar, M. J., and Snyder, S. H., 1976, Opiate receptors: Autoradiographic localization in rat brain, Proc. Natl. Acad. Sci. USA 73:3729–3733.PubMedCrossRefGoogle Scholar
  52. Pollard, H., Llorens, C, Schwartz, J. C., Gros, C, and Dray, F., 1978, Localization of opiate receptors and enkephalins in the rat striatum in relationship with nigrostriatal dopaminergic system: Lesion studies, Brain Res. 151:392–398.PubMedCrossRefGoogle Scholar
  53. Polz-Tejera, G., Schmidt, J., and Karten, H. J., 1975, Autoradiographic localization of α-bungarotoxin-binding sites in the central nervous system, Nature 258:349–351.PubMedCrossRefGoogle Scholar
  54. Porter, C, Barnard, E. A., and Chiu, T. H., 1973, The ultrastructural localization and quantitation of cholinergic receptors at the mouse motor endplate, J. Membr. Biol. 14:383–402.PubMedCrossRefGoogle Scholar
  55. Quirion, R., 1983, Interactions between neurotensin and dopamine in the brain: An overview, Peptides 4:609–615.PubMedCrossRefGoogle Scholar
  56. Quirion, R., Gaudreau, P., St. Pierre, S., Rioux, F., and Pert, C. B., 1982, Autoradiographic distribution of [3H]neurotensin receptors in rat brain: Visualization by tritium sensitive film, Peptides 3:757–763.PubMedCrossRefGoogle Scholar
  57. Reisine, T. D., Nagy, J. I., Beaumont, K., Fibiger, H. C., and Yamamura, H. I., 1979, The localization of receptor binding sites in the substantia nigra and striatum of the rat, Brain Res. 177:241–252.PubMedCrossRefGoogle Scholar
  58. Richards, J. G., Schoch, P., Häring, P., Takacs, B., and Möhler, H., 1987, Resolving GABAA/benzodiazepine receptors: cellular and subcellular localization in the CNS with monoclonal antibodies, J. Neurosci. 7:1866–1886.PubMedGoogle Scholar
  59. Roemer, D., Buescher, H. H., Hill, R. C., Pless, J., Bauer, W., Cardinaux, F., Closse, A., Hauser, D., and Huguenin, R., 1977, A synthetic enkephalin analogue with prolonged parenteral and oral analgesic activity, Nature 268:547–549.PubMedCrossRefGoogle Scholar
  60. Rotter, A., Birdsall, N. J. M., Burgen, A. S. V., Field, P. M., and Raisman, G., 1977, Axotomy causes loss of muscarinic receptors and loss of synaptic contacts in the hypoglossal nucleus, Nature 266:734–735.PubMedCrossRefGoogle Scholar
  61. Sadoul, J. L., Mazella, J., Amar, S., Kitabgi, P., and Vincent, J. P., 1984, Preparation of neurotensin selectively iodinated on the tyrosine 3 residue: Biological activity and binding properties on mammalian neurotensin receptors, Biochem. Biophys. Res. Commun. 120:812–819.PubMedCrossRefGoogle Scholar
  62. Salpeter, M. M., McHenry, F. A., and Salpeter, E. E., 1978, Resolution in electron microscope autoradiography. IV. Application to analysis of autoradiographs, J. Cell Biol. 76:127–145.PubMedCrossRefGoogle Scholar
  63. Schmidt, F. O., 1984, Molecular regulators of brain function: A new view, Neuroscience 13:991–1001.CrossRefGoogle Scholar
  64. Schubert, P., Höllt, V., and Herz, A., 1975, Autoradiographic evaluation of the intracerebral distribution of 3H-etorphine in the mouse brain, Life Sci. 16:1855–1856.PubMedCrossRefGoogle Scholar
  65. Seidah, N. G., Dennis, M., Corvol, R., Rochemont, J., and Chrétien, M., 1980, A rapid high-performance liquid chromatography purification method of iodinated polypeptide hormones, Anal. Biochem. 109:185–191.PubMedCrossRefGoogle Scholar
  66. Snyder, S. H., and Bennett, J. P., Jr., 1976, Neurotransmitter receptors in the brain: Biochemical identification, Annu. Rev. Physiol. 38:153–175.PubMedCrossRefGoogle Scholar
  67. Strader, C. D., Pickel, V. M., Joh, T. H., Strohsacker, M. W., Shorr, R. G. L., Lefkowitz, R. J., and Caron, M. G., 1983, Antibodies to the α-adrenergic receptor: Attenuation of catecholamine-sensitive adenylate cyclase and demonstration of postsynaptic receptor localization in brain, Proc. Natl. Acad. Sci. USA 80:1840–1844.PubMedCrossRefGoogle Scholar
  68. Szigethy, E., Kitabgi, P., and Beaudet, A., 1986, Localization of neurotensin binding sites to dopaminergic and putative cholinergic neurons in rat central nervous system, Soc. Neurosci. Abstr. 12:1003.Google Scholar
  69. Trémeau, O., Faure, G., Boulain, J. C., Bouet, F., Ménez, A., Lecocq, G., Morgat, J. L., and Fromageot, P.,1981, Liaison d’un analogue enképhalinergique (FK 33–824) tritié à une fraction mitochondriale de cerveau de rat, Biochimie 63:477–484.PubMedCrossRefGoogle Scholar
  70. Triller, A., Cluzeaud, F., Pfeiffer, F., Betz, H., and Korn, H., 1985, Distribution of glycine receptors at central synapses: An immunoelectron microscopy study, J. Cell Biol. 101:683–688.PubMedCrossRefGoogle Scholar
  71. Uhl, G. R., 1982, Distribution of neurotensin and its receptors in the central nervous system, Ann. N.Y. Acad. Sci. 400:132–149.PubMedCrossRefGoogle Scholar
  72. Van Loon, G. R., and George, S. R., 1983, Uptake/internalization of met-enkephalin by brain synaptosomes, Life Sci. 33:145–148.PubMedCrossRefGoogle Scholar
  73. Williams, M. A., 1969, The assessment of electron microscopic autoradiography, in: Advances in Optical and Electron Microscopy, Vol. 3 (R. Barer and V. E. Cosslett, eds.), Academic Press, New York, pp. 219–272.Google Scholar
  74. 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.PubMedCrossRefGoogle Scholar
  75. Young, W. S., III, and Kuhar, M. J., 1979b, Neurotensin receptors: Autoradiographic localization in rat CNS, Eur. J. Pharmacol. 59:161–163.PubMedCrossRefGoogle Scholar
  76. Young, W. S., III, and Kuhar, M. J., 1981, Neurotensin receptor localization by light microscopic autoradiography in rat brain, Brain Res. 206:272–285.CrossRefGoogle Scholar
  77. Young, W. S., Wamsley, J. K., Zarbin, M. A., and Kuhar, M. J., 1980, Opioid receptors undergo axonal transport, Science 210:76–77.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • A. Beaudet
    • 1
  • E. Hamel
    • 1
  • K. Leonard
    • 1
  • M. Vial
    • 2
  • E. Moyse
    • 1
  • P. Kitabgi
    • 3
  • J. P. Vincent
    • 3
  • W. Rostène
    • 2
  1. 1.Montreal Neurological InstituteMcGill UniversityMontrealCanada
  2. 2.INSERM U-55, Hôpital St. AntoineParisFrance
  3. 3.Centre de Biochimie du CNRSUniversité de NiceNice CedexFrance

Personalised recommendations