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Visualization and Counting of Receptors at the Light and Electron Microscope Levels

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General Principles and Procedures

Part of the book series: The Receptors ((REC,volume 1))

Abstract

Material elsewhere in this volume provides ample documentation of the existence of specific receptor molecules for a variety of signal-transmitting molecules acting on cells. The neurotransmitters and the circulating hormones are the most obvious categories of these messengers (although far from the only ones). We shall use the term receptor here to denote such structures in the cell plasma membrane, equipped specifically to recognize primary messenger molecules arriving at their cell and to trigger directly the cellular response to these or to initiate the required signal to other machinery that generates the response (e.g., a separate nucleotide cyclase). In contrast, messenger-binding components in the cell interior which are not concerned with the primary recognition of externally incident messengers, while they may be involved in some cases in mediating the cellular response (e.g., to thyroid or steroid hormones), present an entirely different aspect to the problem of receptor location and will not be considered here. Receptors involved in the immune response, which are a very special case, are also outside the scope of this chapter.

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References

  • Abel, J. H., 1979, Sites of binding and metabolism of gonadotropic hormones in the mammalian ovary, in: Drugs, Hormones and Membranes (R. W. Straub, ed.), Raven Press, New York, in press.

    Google Scholar 

  • Albuquerque, E. X., Barnard, E. A., Jansson, S.-E., and Wieckowski, J., 1973, Occupancy of the cholinergic receptors in relation to changes in the end plate potential, Life Sci. 12:545.

    Article  PubMed  CAS  Google Scholar 

  • Albuquerque, E. X., Barnard, E. A., Porter, C. W„ and Warnick, J. E., 1974, The density of acetylcholine receptors and their sensitivity in the postsynaptic membrane of muscle endplates, Proc. Natl Acad. Sci. USA 71:2818

    Article  PubMed  CAS  Google Scholar 

  • Alexander, R. W., Davis, J. N., and Lefkowitz, R. J., 1975, Direct identification and characterization of β-adrenergic receptors in rat brain, Nature (London) 358:437.

    Article  Google Scholar 

  • Anderson, M. J., and Cohen, M. W., 1974, Fluorescent staining of acetylcholine receptors in vertebrate skeletal muscle, J. Physiol. (London) 237:385.

    CAS  Google Scholar 

  • Anderson, M. J., and Cohen, M. W., 1977, Nerve-induced and spontaneous redistribution of acetylcholine receptors in cultured muscle cells, J. Physiol. (London) 268:757.

    CAS  Google Scholar 

  • Anderson, M. J., Cohen, M. W., and Zorychta, E., 1977, Effects of innervation on the distribution of acetylcholine receptors of cultured muscle cells, J. Physiol. (London) 268:731.

    CAS  Google Scholar 

  • Appleton, T. C., 1966, Resolving power, sensitivity and latent image fading of soluble-compound autoradiographs, J. Histochem. Cytochem. 14:414.

    Article  PubMed  CAS  Google Scholar 

  • Ashitaka, Y., Tsong, Y. Y., and Koide, S. S., 1973, Distribution of tritiated human chorionic gonadotropin in superovulated rat ovary, Proc. Soc. Exp. Biol. Med. 142:395.

    PubMed  CAS  Google Scholar 

  • Atlas, D., and Levitzki, A., 1978, Fluorescent visualization of ß-adrenergic receptors on cell surfaces, FEBS Lett. 135:347.

    Google Scholar 

  • Atlas, D., and Segal, M., 1977, Simultaneous visualization of noradrenergic fibres and β-adreno-receptors in pre- and postsynaptic regions in rat brain, Brain Res. 135:347.

    Article  PubMed  CAS  Google Scholar 

  • Atlas, D., Steer, M. L. and Levitzki, A., 1976, Affinity label for β-adrenergic receptor in turkey erythrocytes, Proc. Natl. Acad. Sci. USA 73:1921.

    Article  PubMed  CAS  Google Scholar 

  • Atlas, D., Hanski, E., and Levitzki, A., 1977, Eighty thousand β-adrenoreeeptors in a single cell, Nature (London) 268:144.

    Article  CAS  Google Scholar 

  • Atweh, S. F., and Kuhar, M. J., 1977, Autoradiographic localization of opiate receptors in rat brain. III. The telencephalon, Brain Res. 134:393.

    Article  PubMed  CAS  Google Scholar 

  • Avrameas, S., and Ternynck, T., 1971, Peroxidase-labelled antibody and Fab conjugates with enhanced intracellular penetration, Immunochemistry 8:1175.

    Article  PubMed  CAS  Google Scholar 

  • Axelrod, D., Ravdin, P., Koppel, D. E., Schlessinger, J., Webb, W. W., Elson, E. L., and Podleski, T. R., 1976, Lateral motion of fluorescently labeled acetylcholine receptor in membranes of developing muscle fibers, Proc. Natl. Acad. Sci. USA 73:4594.

    Article  PubMed  CAS  Google Scholar 

  • Barnard, E. A., 1970, Location and measurement of enzymes in single cells by isotopic methods. I, Int. Rev. Cytol. 29:213.

    Article  PubMed  CAS  Google Scholar 

  • Barnard, E. A., Wieckowski, T., and Chiu, T. H., 1971, Cholinergic receptor molecules and Cholinesterase molecules at mouse skeletal muscle junctions, Nature (London) 234:207.

    Article  CAS  Google Scholar 

  • Barnard, E. A., Dolly, J. O., Porter, C. W., and Albuquerque, E. X., 1975, The acetylcholine receptor and the ionic conductance modulation system of skeletal muscle, Exp. Neurol. 48:1.

    Article  PubMed  CAS  Google Scholar 

  • Barnard, E. A., Coates, V., Dolly, J. O., and Mallick, B., 1977, Binding of α-bungarotoxin and cholinergic ligands to acetylcholine receptors in the membrane of skeletal muscle, Cell Biol. Int. Rep. 1:99.

    Article  PubMed  CAS  Google Scholar 

  • Barnard, E. A., Dolly, J. O., Lang, B., Lo, M., and Shorr, R. G., 1979, The application of specifically-acting toxins to the detection of functional components common to peripheral and central synapses, in: Advances in Cytopharmacology, Vol. 3 (B. Ceccarelli and F. Clementi, eds.), Raven Press, New York, in press.

    Google Scholar 

  • Barofsky, A. L., Lipner, H. J., and Armstrong, D. T., 1971, Localization of radioactivity in immature rat ovaries following physiological doses of 125I-labelled bovine luteinizing hormone, Proc. Soc. Exp. Biol. Med. 138:1062.

    PubMed  CAS  Google Scholar 

  • Bender, A. N., Ringel, S. P., Engel, W. K., Vogel, Z., and Daniels, M. P., 1976, Immunoperoxidase localization of a-bungarotoxin, Ann. NY Acad. Sci. 274:20.

    Article  PubMed  CAS  Google Scholar 

  • Bergeron, J. J. M, Levine, G., Sikstrom, R., O’Shaughnessy, D., Kopriwa, B., Nadler, N. J., and Posner, B. I., 1977, Polypeptide hormone binding sites in vivo: Initial localization of 125I-labeled insulin to hepatocyte plasmalemma as visualized by electron microscope radioautography, Proc. Natl. Acad. Sci. USA 74:5051.

    Article  PubMed  CAS  Google Scholar 

  • Bevan, S., and Steinbach, J. H., 1977, The distribution of α-bungarotoxin binding sites in mammalian skeletal muscle developing in vivo, J. Physiol. (London) 267:195.

    CAS  Google Scholar 

  • Bigbee, J. W., Kosek, J. C., and Eng, L. E., 1977, Effects of primary antiserum dilution on staining “antigen-rich” tissues with the peroxidase-antiperoxidase technique, J. Histochem. Cytochem. 25:443.

    Article  PubMed  CAS  Google Scholar 

  • Blackett, N. M, and Parry, D. M., 1973, A new method for analysing electron microscope auto-radiographs using hypothetical grain distributions, J. Cell. Biol. 57:9.

    Article  PubMed  CAS  Google Scholar 

  • Blackett, N. M., and Parry, D. M., 1977, A simplified method of “hypothetical grain” analysis of electron microscope autoradiographs, J. Histochem. Cytochem. 25:206.

    Article  PubMed  CAS  Google Scholar 

  • Bourgeois, J. P., Tsuji, S., Boquet, P., Pillot, J., Ryter, A., and Changeux, J. P., 1971, Localization of the cholinergic receptor protein by immunofluorescence in eel electroplax, FEBS Lett. 16:92.

    Article  PubMed  CAS  Google Scholar 

  • Bourgeois, J. P., Ryter, A., Menez, A., Fromageot, P. Boquet, and Changeux, J. P., 1972, Localization of the cholinergic receptor protein in Electrophorus electroplax by high resolution autoradiography, FEBS Lett. 25:127.

    Article  PubMed  CAS  Google Scholar 

  • Bourgeois, J. P., Popot, J. L., Ryter, A., and Changeux, J. P., 1973, Consequences of denervation on the distribution of the cholinergic (nicotinic) receptor sites from Electrophorus electricus revealed by high resolution autoradiography, Brain Res. 62:557.

    Article  PubMed  CAS  Google Scholar 

  • Braten, T., 1976, Detection of concanavalin A sites on the surface of algal gametes and zygotes with the scanning electron microscope, J. Ultrastruct. Res. 57:226.

    Google Scholar 

  • Burgen, A. S. V., Hiley, C. R., and Young, J. M., 1974, The properties of muscarinic receptors in mammalian cerebral cortex, Br. J. Pharmacol. 51:279.

    Article  PubMed  CAS  Google Scholar 

  • Èursztajn, S., and Gershon, M. D., 1977, Discrimination between nicotinic receptors in vertebrate ganglia and skeletal muscle by alpha-bungarotoxin and cobra venoms, J. Physiol. (London) 269:17.

    Google Scholar 

  • Burt, D. R., Creese, I., and Snyder, S. H., 1976, Properties of [3H]haloperidol and [3H]dopamine binding associated with dopamine receptors in calf brain membranes, Mol. Pharmacol. 12:800.

    PubMed  CAS  Google Scholar 

  • Bylund, D. B., Charness, M. E., and Snyder, S. H., 1977, Beta adrenergic receptor labelling in intact animals with 125I-hydroxybenzylpindolol, J. Pharmacol. Exp. Ther. 201:644.

    PubMed  CAS  Google Scholar 

  • Castel, M., 1978, Immunocytochemical evidence for vasopressin receptors, J. Histochem. Cytochem. 26:581.

    Article  PubMed  CAS  Google Scholar 

  • Chandler, J. A., 1975, Electron probe x-ray microanalysis in cytochemistry, in: Techniques of Biochemical and Biophysical Morphology, Vol. 2 (D. Glick and R. M. Rosenbaum, eds.), pp. 307–438, Wiley, New York.

    Google Scholar 

  • Chang, C. M., Reitherman, R. W., Rosen, S. D., and Barondes, S. H., 1975, Cell surface location of discoidin, a developmentally regulated carbohydrate binding protein from Dictyostelium dis-coideum, Exp. Cell Res. 95:136.

    Article  PubMed  CAS  Google Scholar 

  • Changeux, J. P., 1975, The cholinergic receptor protein from fish electric organ, in: Handbook of Psychopharmacology, Vol. 6 (L. L. Trepon, S. D. Trepon, and S. H. Snyder, eds.), pp. 235–301, Plenum Press, New York.

    Google Scholar 

  • Changeux, J. P., Benedetti, L., Bourgeois, J. P., Brisson, A., Cartaud, J., Devaux, P., Grünhagen, H., Moreau, M., Popot, J. L., Sobel, A., and Weber, M., 1975, Some structural properties of the cholinergic receptor protein in its membrane environment relevant to its function as a pharmacological receptor, Cold Spring Harbor Symp. Quant. Biol. 25:211.

    Google Scholar 

  • Chiu, T. H., Dolly, J. O., and Barnard, E. A., 1973, Solubilization from skeletal muscle of two components that specifically bind a-bungarotoxin, Biochem. Biophys. Res. Commun. 51:205.

    Article  PubMed  CAS  Google Scholar 

  • Cohen, J. B., and Changeux, J. P., 1975, The cholinergic receptor protein in its membrane environment, Annu. Rev. Pharmacol. 15:83.

    Article  PubMed  CAS  Google Scholar 

  • Conti-Tronconi, B. M„ Gotti, C., and Clementi, F., 1978, Molecular organisation of cholinergic receptor in post-synaptic membrane of Torpedo marmurata, in: The Biochemistry of Myasthenia Gravis and Muscular Distrophy (G. G. Lunt and R. M. Marchbanks, eds.), pp. 111–119, Academic Press, New York.

    Google Scholar 

  • Coons, A. H., 1956, Histochemistry with labelled antibody, Int. Rev. Cytol. 5:1.

    Article  CAS  Google Scholar 

  • Craig, S. W., and Cuatrecasas, P. S., 1975, Mobility of cholera toxin receptors on rat lymphocyte membranes, Proc. Natl. Acad. Sci. USA 72:3844.

    Article  PubMed  CAS  Google Scholar 

  • Creese, R., and MacLagan, J., 1970, Entry of decamethonium in rat muscle studied by autoradiography, J. Physiol. (London) 210:363.

    CAS  Google Scholar 

  • Cuatrecasas, P., 1973, Gangliosides and membrane receptors for cholera toxin, Biochemistry 12:3558.

    Article  PubMed  CAS  Google Scholar 

  • Cuatrecasas, P., and Hollenberg, M. D., 1976, Membrane receptors and hormone action, Adv. Protein Chem. 30:251.

    Article  PubMed  CAS  Google Scholar 

  • Daniels, M. P., and Vogel, Z., 1975, Immunoperoxidase staining of a-bungarotoxin binding sites in muscle endplates shows distribution of acetylcholine receptors, Nature (London) 254:339.

    Article  CAS  Google Scholar 

  • Da Silva, P. and Branton, D., 1970, Membrane splitting in freeze-etching. Covalently bound ferritin as a membrane marker, J. Cell Biol. 45:598.

    Article  Google Scholar 

  • Da Silva, P., Moss, P. S., and Fudenberg, H. H., 1973, Anionic sites on the membrane-intercalated particles of human erythrocyte ghost membranes: Freeze-etch localization, Exp. Cell Res. 81:127.

    Article  Google Scholar 

  • De Pétris, S., and Raff, M. C., 1973, Normal distribution, patching and capping of lymphocyte surface immunoglobulin studied by electron microscopy, Nature (London) New Biol. 241:257.

    Google Scholar 

  • Diab, I. M., Freedman, D. X., and Roth, L. J., 1971, 3H-lysergic acid diethylamide: Cellular autoradiographic localization in rat brain, Science 173:1022.

    Google Scholar 

  • DiPasquale, A., Varga, J. M., Moellman, G., and McGuire, J. S., 1978, Synthesis of a hormonally active conjugate of a-MSH, ferritin and fluorescein, Anal. Biochem. 84:37.

    Article  PubMed  CAS  Google Scholar 

  • Drachman, D. B., Kao, I., Pestronk, A., and Toyka, K. V., 1976, Myasthenia gravis as a receptor disorder, Ann. NY Acad. Sci. 274:226.

    Article  PubMed  CAS  Google Scholar 

  • Eide, R., Hökfelt, T., Johnansson, O., and Terenius, L., 1976, Immunohistochemical studies using antibodies to leucine-enkephalin: Initial observations on the nervous system of the rat, Neuroscience 1:349.

    Article  Google Scholar 

  • Engel, A. G., Lindstrom, J. M., Lambert, E. H., and Lennon, V. A., 1977, Ultrastructural localization of the acetylcholine receptor in myasthenia gravis and in its experimental autoimmune model, Neurology 27:307.

    Article  PubMed  CAS  Google Scholar 

  • Enna, S. J., and Snyder, S. H., 1977, Influences of ions, enzymes and detergents on α-aminobutyric acid binding in synaptic membranes of rat brain, Mol. Pharmacol. 13:442.

    PubMed  CAS  Google Scholar 

  • Fahimi, H. D., 1975, Fine-structural cytochemical localization of peroxidatic activity of catalase, in: Techniques of Biochemical and Biophysical Morphology, Vol. 2 (D. Glick and R. M. Rosenbaum, eds.), pp. 197–246, Wiley, New York.

    Google Scholar 

  • Fambrough, D. M., 1974, Acetylcholine receptors. Revised estimates on extrajunctional receptor density in denervated rat diaphragm, J. Gen. Physiol. 64:468.

    Article  PubMed  CAS  Google Scholar 

  • Fedinec, A. F., Debrow, M. M., Gardner, D. P., and Sternberger, L. A., 1970, Immunohistochemical localization of tetanus toxin by the unlabeled antibody enzyme method, J. Histochem. Cytochem. 18:684.

    Google Scholar 

  • Fertuck, H. C., and Salpeter, M. M., 1974a, Sensitivity in electron microscope autoradiography for 125I, J. Histochem. Cytochem. 22:80.

    Article  CAS  Google Scholar 

  • Fertuck, H. C., and Salpeter, M. M., 1974b, Localization of acetylcholine receptor by 125I-labelled α-bungarotoxin binding at mouse motor endplates, Proc. Natl. Acad. Sci USA 71:1376.

    Article  CAS  Google Scholar 

  • Fertuck, H. C., and Salpeter, M. M., 1976, Quantitation of junctional and extra-junctional acetylcholine receptors by electron microscope autoradiography after 125I-α-bungarotoxin binding at mouse neuromuscular junctions, J. Cell Biol. 69:144.

    Article  PubMed  CAS  Google Scholar 

  • Fisher, K. A., and Branton, D., 1976, Freeze-fracture autoradiography: Feasibility, J. Cell Biol. 70:453.

    Article  PubMed  CAS  Google Scholar 

  • Freeman, J. A., 1977, Possible regulatory function of acetylcholine receptor in maintenance of retino-tectal synapses, Nature (London) 269:218.

    Article  CAS  Google Scholar 

  • Gerlach, J. L., and McEwen, B. S., 1972, Rat brain binds adrenal steroid hormone: Radioautography of hippocampus with corticosterone, Science 175:1133.

    Article  PubMed  CAS  Google Scholar 

  • Goldfine, I. D., Smith, G. J., Wong, K. Y., and Jones, A. L., 1977, Cellular uptake and nuclear binding of insulin in human cultured lymphocytes: Evidence for potential intracellular sites of insulin action, Proc. Natl. Acad. Sci. USA 74:1368.

    Article  PubMed  CAS  Google Scholar 

  • Gonatas, N. K., Stieber, A., Gonatas, J., Gambetti, P., Antoine, J. C., and Avarameas, S., 1974, Ultrastructural autoradiographic detection of intracellular immunoglobulins with iodinated Fab fragments of antibody: The combined use of ultrastructural autoradiography and peroxidase cytochemistry for the detection of two antigens (double labelling), J. Histochem. Cytochem. 22:999.

    Article  PubMed  CAS  Google Scholar 

  • Gould, R. M, and Dawson, R. M. C., 1976, Incorporation of newly formed lecithin into purified nerve myelin, J. Cell Biol. 68:480.

    Article  PubMed  CAS  Google Scholar 

  • Graham, R. C, Jr., and Karnovsky, M. J., 1966, The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: Ultrastructural cytochemistry by a new technique, J. Histochem. Cytochem. 14:291.

    Article  PubMed  CAS  Google Scholar 

  • Hansson, H. A., Holmgren, J., and Svenneholm, L., 1977, Ultrastructural localization of cell membrane GM1 ganglioside by cholera toxin, Proc. Natl. Acad. Sci. USA 74:3782.

    Article  PubMed  CAS  Google Scholar 

  • Hardy, H., and Heimer, L., 1977, A safer and more sensitive substitute for diaminobenzidine in the light microscopic demonstration of retrograde and anterograde axonal transport of horseradish peroxidase, Neurosci. Lett. 5:23.

    Article  Google Scholar 

  • Hartzell, H. C., and Fambrough, D. M., 1972, Acetylcholine receptors: Distribution and extra-junctional density in rat diaphragm after denervation with acetylcholine sensitivity, J. Gen. Physiol. 60:248.

    Article  PubMed  CAS  Google Scholar 

  • Hartzell, H. C., and Fambrough, D. M., 1973, Acetylcholine receptor production and incorporation into membranes of developing muscle fibers, Dev. Biol. 30:153.

    Article  PubMed  CAS  Google Scholar 

  • Hijmans, W., and Schaeffer, M., 1975, Fifth international conference on immunofluorescence and related staining techniques, Ann. NY Acad. Sci. 254:1.

    Article  Google Scholar 

  • Hinton, D. M., Petrali, J. P., Meyer, H. G., and Sternberger, L. A., 1973, The unlabeled antibody enzyme method of immunohistochemistry: Molecular immunocytochemistry of antibodies in the erythrocyte surface, J. Histochem. Cytochem. 21:978.

    Article  PubMed  CAS  Google Scholar 

  • Hodges, G. M., and Muir, M., 1975, Quantitative evaluation of autoradiographs by x-ray spectroscopy, J. Microsc. (Oxford) 104:173.

    Article  CAS  Google Scholar 

  • Hopkins, C. R., and Gregory, H., 1977, Topographical localization of the receptors for luteinizing hormone-releasing hormone on the surface of dissociated pituitary cells, J. Cell Biol. 75:528.

    Article  PubMed  CAS  Google Scholar 

  • Horowitz, S. B., 1974, The ultra-low temperature autoradiography of water and its solutes, Methods Cell Biol. 8:249.

    Article  PubMed  CAS  Google Scholar 

  • Hough, P. V. C., McKinney, W. R., Ledbetter, M. C., Pollack, R. E., and Moos, H. W., 1976, Identification of biological macromolecules in situ at high resolution via the fluorescence excited by a scanning electron beam, Proc. Natl. Acad. Sci. USA 73:317.

    Article  PubMed  CAS  Google Scholar 

  • Hourani, B. T., Torain, B. F., Henkart, M. P., Carter, R. L., Marchesi, V. T., and Fischbach, G. D., 1974, Acetylcholine receptors of cultured muscle cells demonstrated with ferritin-α-bungarotoxin conjugates, J. Cell. Sci. 16:473.

    PubMed  CAS  Google Scholar 

  • Howe, P. R. C., Telfer, J. A., and Austin, L., 1976, Binding sites for 125I-labeled-α-bungarotoxin in normal and denervated mouse muscle, Exp. Neurol. 52:272.

    Article  PubMed  CAS  Google Scholar 

  • Ito, Y., Miledi, R., Vincent, A., and Newsom-Davis, J., 1978, Acetylcholine receptors and end-plate electrophysiology in myasthenia gravis, Brain 101:345.

    Article  PubMed  CAS  Google Scholar 

  • Jarett, L., and Smith, R. D., 1975, Ultrastructural localization of insulin receptors on adipocytes, Proc. Natl. Acad. Sci. USA 72:3526.

    Article  PubMed  CAS  Google Scholar 

  • Jedrzejczyk, J., Rymaszewska, T., Wieckowski, J., and Barnard, E. A., 1973, Dystrophic chicken muscle: Altered synaptic Cholinesterase, Science 180:406.

    Article  PubMed  CAS  Google Scholar 

  • Keefer, D. A., Stumpf, W. E., Petrusz, P., and Sar, M., 1975, Simultaneous autoradiographic and immunohistochemical localization of estrogen and gonadotropin in the rat pituitary, Am. J. Anat. 142:129.

    Article  PubMed  CAS  Google Scholar 

  • Keefer, D. A., Stumpf, W. E., and Petrusz, P., 1976, Quantitative autoradiographic assessment of 3H-estradiol uptake in immunocytochemically characterized pituitary cells, Cell Tissue Res. 166:25.

    Article  PubMed  CAS  Google Scholar 

  • Kendall, P., and Barnard, E. A., 1967, Thiolation for labelling of proteins, using catalysis by silver and imidazole combined, Biochim. Biophys. Acta 188:10.

    Google Scholar 

  • Ko, P. K., Anderson, M. J., and Cohen, M. W., 1977, Denervated skeletal muscle fibers develop discrete patches of high acetylcholine sensitivity, Science 196:540.

    Article  PubMed  CAS  Google Scholar 

  • Kopriwa, B. M., 1967, A semi-automatic instrument for the radioautographic coating technique, J. Histochem. Cytochem. 14:923.

    Article  Google Scholar 

  • Kraehenbuhl, J. P., Racine, L., and Galardy, R. E., 1975, Localization of secretory IgA, secretory component, and α-chain in the mammary gland of lactating rabbits by immunoelectron microscopy, Ann. NY Acad. Sci. 254:190.

    Article  PubMed  CAS  Google Scholar 

  • Kuhar, M. J., and Yamamura, H. I., 1975, Light autoradiographic localization of cholinergic muscarinic receptors in rat brain by specific binding of a potent antagonist, Nature (London) 253:560.

    Article  CAS  Google Scholar 

  • Kuhar, M. J., and Yamamura, H. I., 1976, Localization of cholinergic muscarinic receptors in rat brain by light microscopic radioautography, Brain Res. 110:229.

    Article  PubMed  CAS  Google Scholar 

  • Kühlmann, W., and Avrameas, S., 1971, Glucose oxidase as an antigen marker for light and electron microscopic studies, J. Histochem. Cytochem. 19:361.

    Article  PubMed  Google Scholar 

  • Land, B. R., Podleski, T. R., Salpeter, E. E., and Salpeter, M. M., 1977, Acetylcholine receptor distribution on myotubes in culture correlated to acetylcholine sensitivity, J. Physiol. (London) 269:155.

    CAS  Google Scholar 

  • Lang, B., Barnard, E. A., Cavanagh, J., and Dolly, J. O., 1979, in preparation.

    Google Scholar 

  • Lentz, T. L., Mazurkiewicz, J. F., and Rosenthal, J., 1977, Cytochemical localization of acetylcholine receptors at the neuromuscular junction by means of horseradish peroxidase-labelled α-bungarotoxin, Brain Res. 132:423.

    Article  PubMed  CAS  Google Scholar 

  • Libelius, R., 1974, Binding of 3H-labelled cobra neurotoxin to cholinergic receptors in fast and slow mammalian muscles, J. Neural Transm. 35:137.

    Article  PubMed  CAS  Google Scholar 

  • Libelius, R., 1975, Evidence for endocytotic uptake of cobra neurotoxin in mouse skeletal muscle, J. Neural Transm. 37:161.

    Google Scholar 

  • Libelius, R., Eaker, D., and Karlsson, E., 1975, Further studies on the binding properties of cobra neurotoxin to cholinergic receptors in mouse skeletal muscle, J. Neural Transm. 37:165.

    Article  PubMed  CAS  Google Scholar 

  • Lord, J. A. H., Waterfield, A. A., Hughes, J., and Kosterlitz, H. W., 1977, Endogeneous opioid peptides: Multiple agonists and receptors, Nature (London) 267:495.

    Article  CAS  Google Scholar 

  • Malmgren, L. T., and Olsson, Y., 1977, A more sensitive method for histochemical demonstration of horseradish peroxidase, Proc. Int. Soc. Neurochem. 6:282.

    Google Scholar 

  • Manuelidis, L., and Manuelidis, E. E., 1976, Cholera toxin-peroxidase: Changes in surface labelling of glioblastoma cells with increased time in tissue culture, Science 193:588.

    Article  PubMed  CAS  Google Scholar 

  • Mason, T. C., Phifer, R. F., Spicer, S. S., Swallow, R. A., and Dreshin, R. B., 1969, An immunoglobulin-enzyme bridge method for localizing tissue antigens, J. Histochem. Cytochem. 17:563.

    Article  PubMed  CAS  Google Scholar 

  • McLean, I. W., and Nakane, P. K., 1974, Periodate-lysine-paraformaldehyde fixative: A new fixative for immunoelectron microscopy, J. Histochem. Cytochem. 22:1077.

    Article  PubMed  CAS  Google Scholar 

  • Miledi, R., and Szcepaniak, A., 1975, Effect of Dendroaspis neurotoxin on synaptic transmission in the spinal cord of the frog, Proc. Roy. Soc. London Ser. B 190:267.

    Article  CAS  Google Scholar 

  • Moriarty, G. C., Moriarty, C. M., and Sternberger, L. A., 1973, Ultrastructural immunocytochemistry with unlabelled antibodies and the peroxidase-antiperoxidase complex: A technique more sensitive than radioimmunoassay, J. Histochem. Cytochem. 21:825.

    Article  PubMed  CAS  Google Scholar 

  • Nakane, P. K., 1975, Recent progress in the peroxidase-labeled antibody method, Ann. NY Acad. Sci. 254:203.

    Article  PubMed  CAS  Google Scholar 

  • Nakane, P. K., and Kawaoi, A., 1974, Peroxidase-labelled antibody. A new method of conjugation, J. Histochem. Cytochem. 22:1084.

    Article  PubMed  CAS  Google Scholar 

  • Nakane, P. K., and Pierce, G. B., 1966, Enzyme-labeled antibodies: preparation and application for localization of antigens, J. Histochem. Cytochem. 14:929.

    Article  PubMed  CAS  Google Scholar 

  • Nemanic, M. K., Shannon, J. M., Carter, D. P., and Wofsy, L., 1974, Immunospecific labeling of a cell surface antigen with a marker visible in the scanning electron microscope, J. Histochem. Cytochem. 22:295.

    Google Scholar 

  • Novikoff, A. B., Novikoff, P. M., Quintana, N., and Davis, C., 1972, Diffusion artefact in 3,3′-diaminobenzidine cytochemistry, J. Histochem. Cytochem. 20:745.

    Article  PubMed  CAS  Google Scholar 

  • Nowak, T. P., Haywood, P. L., and Barondes, S. H., 1976, Developmentally regulated lectin in embryonic chick muscle and a myogenic cell line, Biochem. Biophys. Res. Commun. 68:650.

    Article  PubMed  CAS  Google Scholar 

  • Olsen, R. W., Lee, J. M., and Sar, M., 1975, Binding of α-aminobutyric acid to crayfish muscle and its relationship to receptor sites, Mol. Pharmacol. 11:566.

    PubMed  CAS  Google Scholar 

  • Orth, J., and Christensen, A. K., 1977, Localization of 125I-labelled FSH in the testes of hypophysectomized rats by autoradiography at the light and electron microscope levels, Endocrinology 101:262.

    Article  PubMed  CAS  Google Scholar 

  • Patrick, J., and Stallcup, W. B., 1977, Immunological distinction between acetylcholine receptor and the α-bungarotoxin-binding component on sympathetic neurons, Proc. Natl. Acad. Sci. USA 74:4689.

    Article  PubMed  CAS  Google Scholar 

  • Pearse, A. G. E., 1972, Histochemistry, Theoretical and Applied, Vol. 2, 3rd ed., Churchill, London.

    Google Scholar 

  • Pert, C. B., Kuhar, M. J., and Snyder, S. H., 1975, Autoradiographic localization of the opiate receptor in rat brain, Life Sci. 16:1849.

    Article  PubMed  CAS  Google Scholar 

  • Pert, C. B., Kuhar, M. J., and Snyder, S. H., 1976, Opiate receptor: Autoradiographic localization in rat brain, Proc. Natl. Acad. Sci. USA 73:3729.

    Article  PubMed  CAS  Google Scholar 

  • Peters, P. D., Yarom, R., Dorman, A., and Hall, T., 1976, X-ray microanalysis of intracellular zinc: EMMA-4 examinations of normal and injured muscle and myocardium, J. Ultrastruct. Res. 57:121.

    Article  PubMed  CAS  Google Scholar 

  • Peters, R., Peters, J., Tews, K. H., and Bahr, W., 1974, A microfluorimetric study of translational diffusion in erythrocyte membranes. Biochim. Biophys. Acta 367:282.

    Article  PubMed  CAS  Google Scholar 

  • Petrali, J. P., Hinton, D. M., Moriarty, G. C., and Sternberger, L. A., 1974, The unlabeled antibody enzyme method of immunocytochemistry: Quantitative comparison of sensitivities with and without peroxidase-antiperoxidase complex, J. Histochem. Cytochem. 22:782.

    Article  PubMed  CAS  Google Scholar 

  • Petrusz, P., 1974, Demonstration of gonadotropin-binding sites in the rat ovary, by an immunoglobulin-enzyme bridge method, Eur. J. Obstet. Gynecol. Reprod. Biol. Suppl. 4:53.

    Google Scholar 

  • Petrusz, P., 1975, Localization and sites of action of gonadotropins in brain, in: Anatomical Neuroendocrinology (W. E. Stumpf and L. D. Grant, eds.), pp. 176–184, Karger, Basel.

    Google Scholar 

  • Petrusz, P., 1979, Gonadotropin receptors, in: Drugs, Hormones and Receptors (R. W. Straub, ed.), Raven Press, New York, in press.

    Google Scholar 

  • Petrusz, P., Sar, M., Ordronneau, P., and Di Meo, P., 1976, Specificity in immunocytochemical staining, J. Histochem. Cytochem. 24:1110.

    Article  PubMed  CAS  Google Scholar 

  • Ploem, J. S., 1975, in Hijmans and Schaeffer, 1975, pp. 4–20.

    Google Scholar 

  • Polz-Tejera, G., Schmidt, J., and Karten, H. J., 1975, Autoradiographic localization of α-bungarotoxin binding sites in the CNS, Nature (London) 258:349.

    Article  CAS  Google Scholar 

  • Porter, C. W., and Barnard, E. A., 1975a, The density of cholinergic receptors at the endplate postsynaptic membrane: Ultrastructural studies in two mammalian species, J. Membr. Biol. 20:31.

    Article  CAS  Google Scholar 

  • Porter, C. W., and Barnard, E. A., 1975b, Distribution and density of cholinergic receptors at the motor endplates of a denervated mouse muscle, Exp. Neurol. 48:542.

    Article  CAS  Google Scholar 

  • Porter, C. W., and Barnard, E. A., 1976, Ultrastructural studies on the acetylcholine receptor at motor endplates of normal and pathologic muscles, Ann. NY Acad. Sci. 274:85.

    Article  PubMed  CAS  Google Scholar 

  • Porter, C. W., Chiu, T. H., Wieckowski, J., and Barnard, E. A., 1973a, Types and locations of cholinergic receptor-like molecules in muscle fibres, Nature (London) New Biol. 241:3.

    CAS  Google Scholar 

  • Porter, C. W., Barnard, E. A., and Chiu, T. H., 1973b, The ultrastructural localization and quantitation of cholinergic receptors at the mouse motor endplate, J. Membr. Biol. 14:383.

    Article  CAS  Google Scholar 

  • Price, D. L., Griffin, J. W., and Peck, J., 1977, Tetanus toxin: Evidence for binding at presynaptic nerve endings, Brain Res. 121:379.

    Article  PubMed  CAS  Google Scholar 

  • Prives, J., Silman, I., and Amsterdam, A., 1976, Appearance and disappearance of ACh receptors during differentiation of chick skeletal muscle in vitro, Cell 7:543.

    Article  PubMed  CAS  Google Scholar 

  • Rajaniemi, H., and Vanha-Perttula, T., 1973, Attachment to the luteal plasma membranes: An early event in the action of luteinizing hormone, J. Endocrinol. 57:199.

    Article  PubMed  CAS  Google Scholar 

  • Ravdin, P., and Axelrod, D., 1977, Fluorescent tetramethyl-rhodamine derivatives of α-bungarotoxin: Preparation, separation and characterization, Anal. Biochem. 80:585.

    Article  PubMed  CAS  Google Scholar 

  • Ringel, S. P., Bender, A. N., Festoff, B. W., Engel, W. K., Vogel, Z., and Daniels, M. P., 1975, Ultra-structural demonstration and analytical application of extrajunctional receptors of denervated human and rat skeletal muscle fibres, Nature (London) 255:730.

    Article  CAS  Google Scholar 

  • Ritchie, J. M., and Rogart, R. B., 1977, The binding of labelled saxitoxin to the sodium channels in normal and denervated mammalian muscle, and in amphibian muscle, J. Physiol. (London) 269:341.

    CAS  Google Scholar 

  • Rogers, A. W., 1967, Techniques of Autoradiography, Elsevier, Amsterdam.

    Google Scholar 

  • Rosenbluth, J., 1972, Myoneural junctions of two ultrastructurally distinct types in earthworm body wall, J. Cell Biol. 54:566.

    Article  PubMed  CAS  Google Scholar 

  • Rosenbluth, J., 1974, Substructure of amphibian motor endplate, J. Cell Biol. 62:755.

    Article  PubMed  CAS  Google Scholar 

  • Ross, M. J., Klymkowsky, M. W., Agard, D. A., and Stroud, R. M, 1977, Structural studies of a membrane-bound acetylcholine receptor from Torpedo californica, J. Mol. Biol. 116:635.

    Article  CAS  Google Scholar 

  • Rotter, A., Birdsall, N. J. M., Burgen, A. S. V., Field, P. M., and Raisman, G., 1977a, Axotomy causes loss of muscarinic receptors and loss of synaptic endplates in the hypoglossal nucleus, Nature (London) 266:734.

    Article  CAS  Google Scholar 

  • Rotter, A., Birdsall, N. J. M., Field, P. M., and Raisman, G., 1977b, personal communication.

    Google Scholar 

  • Salpeter, M. M., and Szabo, M., 1972, Sensitivity in electron microscope autoradiography. I. The effect of radiation dose, J. Histochem. Cytochem. 20:425.

    Article  PubMed  CAS  Google Scholar 

  • Salpeter, M. M., and Szabo, M., 1976, An improved Kodak emulsion for use in high resolution electron microscope autoradiography, J. Histochem. Cytochem. 24:1204.

    Article  PubMed  CAS  Google Scholar 

  • Salpeter, M. M., Bachmann, L., and Salpeter, E. E., 1969, Resolution in electron microscope autoradiography, J. Cell Biol. 41:1.

    Article  PubMed  CAS  Google Scholar 

  • Salpeter, M. M., Fertuck, H. C., and Salpeter, E. E., 1977, Resolution in electron microscope autoradiography, III. Iodine-125, the effect of heavy metal staining and reassessment of critical parameters, J. Cell Biol. 72:161.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Schurer, J. W., Hoedemaeker, P. J., and Molenaar, I., 1977, Polyethyleneimine as tracer particle for (immuno)electron microscopy, J. Histochem. Cytochem. 25:384.

    Article  PubMed  CAS  Google Scholar 

  • Seligman, A. M., Shannon, W. A., Jr., Hoshino, Y., and Plopinger, R. E., 1973, Some important principles in 3,3′-diaminobenzidine ultrastructural cytochemistry, J. Histochem. Cytochem. 21:756.

    Article  PubMed  CAS  Google Scholar 

  • Shain, W., Green, L. A., Carpenter, D. O., Sytkowski, A. J., and Vogel, Z., 1974, Aplysia acetylcholine receptors: Blockade by and binding of a-bungarotoxin, Brain Res. 72:225.

    Article  PubMed  CAS  Google Scholar 

  • Shannon, L. M., Kay, E., and Lew, J. Y., 1966, Peroxidase isoenzymes from horseradish roots. I. Isolation and physical properties, J. Biol. Chem. 241:2166.

    PubMed  CAS  Google Scholar 

  • Shotton, D. M., Heuser, J. E., Reese, B. F., and Reese, T. S., 1979, Postsynaptic membrane folds of the frog neuromuscular junction visualized by scanning electron microscopy, Neuroscience, in press.

    Google Scholar 

  • Siess, E., Wieland, O., and Miller, F., 1971, A simple method for the preparation of pure and active a-globulin-ferritin conjugates using glutaraldehyde, Immunology 20:659.

    PubMed  CAS  Google Scholar 

  • Simantov, R., Kuhar, M. J., and Snyder, S. H., 1977, Opioid peptide enkephalins: Immunohisto-chemical mapping in the rat central nervous system, Proc. Natl. Acad. Sci. USA 74:2167.

    Article  PubMed  CAS  Google Scholar 

  • Singer, S. J., 1976, The fluid mosaic model of membrane structure: some applications to ligand-receptor and cell-cell interactions, in: Surface Membrane Receptors (R. A. Bradshaw, W. A. Frazier, M. C. Merrell, D. I. Gottlieb, and R. A. Hogue-Angeletti, eds.), pp. 1–24, Plenum Press, New York.

    Chapter  Google Scholar 

  • Singer, S. J., and Nicolson, G. L., 1972, The fluid mosaic model of the structure of cell membranes, Science 175:720.

    Article  PubMed  CAS  Google Scholar 

  • Singer, S. J., and Schick, A. F., 1961, The properties of specific stains for electron microscopy prepared by the conjugation of antibody molecules with ferritin, J. Biophys. Biochem. Cytol. 9:519.

    Article  PubMed  CAS  Google Scholar 

  • Soloff, M. S., Rees, H. D., Sar, M., and Stumpf, W. E., 1975, Autoradiographic localization of radioactivity from [3H]oxytocin in the rat mammary gland and oviduct, Endocrinology 96:1475.

    Article  PubMed  CAS  Google Scholar 

  • Soni, S. C., Kalnins, V. T., and Haggis, G. H., 1975, Localisation of caps on mouse lymphocytes by scanning electron microscopy, Nature (London) 255:717.

    Article  CAS  Google Scholar 

  • Sternberger, L. A., 1972, The unlabelled-antibody-peroxidase and the quantitative-immunouranium methods in light and electron immunohistochemistry, in: Techniques of Biochemical and Biophysical Morphology, Vol. 1 (D. Glick and R. M. Rosenbaum, eds.), pp. 57–88, Wiley, New York.

    Google Scholar 

  • Sternberger, L. A., 1978, Receptors for luteinizing hormone-releasing hormone, J. Histochem. Cytochem. 26:542.

    Article  PubMed  CAS  Google Scholar 

  • Stockert, R. J., Morell, A. G., and Scheinberg, I. H., 1974, Mammalian hepatic lectin, Science 186:365.

    Article  PubMed  CAS  Google Scholar 

  • Stumpf, W. E., 1970, Tissue preparation for the autoradiographic localization of hormones, in: Introduction to Quantitative Cytochemistry, Vol. 2 (G. L. Wied, and G. F. Bahr, eds.), pp. 507–526, Academic Press, New York.

    Google Scholar 

  • Stumpf, W. E., 1976, Techniques for the autoradiography of diffusible compounds, Methods Cell Biol. 13:171.

    Article  PubMed  CAS  Google Scholar 

  • Stumpf, W. E., and Sar, M., 1977, Steroid hormone target cells in the periventricular brain: relationship to peptide hormone producing cells, Fed. Proc. 36:1973.

    PubMed  CAS  Google Scholar 

  • Suszkiw, J. B., and Ichiki, M, 1976, Fluorescein conjugated a-bungarotoxin: Its properties and interaction with acetylcholine receptors, Anal. Biochem. 73:109.

    Article  PubMed  CAS  Google Scholar 

  • Sytkowski, A. J., Vogel, Z„ and Nirenberg, M. W., 1973, Development of acetylcholine receptor clusters on cultured muscle cells, Proc. Natl. Acad. Sci. USA 70:270.

    Article  PubMed  CAS  Google Scholar 

  • Teichberg, V. I., Silman, I., Beutsch, D. D., and Resheff, G., 1975, A ß-D-galactoside binding protein from electric organ tissue of Electrophorus electricus, Proc. Natl. Acad. Sci. USA 72:1383.

    Article  PubMed  CAS  Google Scholar 

  • Templeton, C. L., Douglas, R. J., and Vail, W. J., 1978, Ferritin conjugated protein A, FEBS Lett. 85:95.

    Article  PubMed  CAS  Google Scholar 

  • Tittler, M., Weinreich, P., and Seeman, P., 1977, New detection of brain dopamine receptors with [3H]dihydroergocryptine, Proc. Natl. Acad. Sci. USA 74:3750.

    Article  PubMed  CAS  Google Scholar 

  • Tsou, K. C., Mela, L., Gupta, P. D., and Lynn, D., 1976, Mitochondrial ultrastructure study with the new cytochrome c binding agent 4–4′-diamino-2–2′-bipyridyl ferrous chelate. J. Ultrastruct. Res. 54:235.

    Article  PubMed  CAS  Google Scholar 

  • Unwin, P. N. T., and Henderson, R., 1975, Molecular structure determination by electron microscopy of unstained crystalline specimens, J. Mol. Biol. 94:425.

    Article  PubMed  CAS  Google Scholar 

  • U’Prichard, D. C., Greenberg, D. A., and Snyder, S. H., 1977, Binding characteristics of a radiolabeled agonist and antagonist at central nervous system alpha-noradrenergic receptors, Mol. Pharmacol. 13:454.

    PubMed  Google Scholar 

  • Van Heyningen, W. E., 1974, Gangliosides as membrane receptors for tetanus toxin, cholera toxin and serotonin, Nature (London) 249:415.

    Article  Google Scholar 

  • Varga, J. M., Saper, M. A., Lerner, A. B., and Fritsch, P., 1976a, Non-random distribution of receptors for melanocyte-stimulating hormones on the surface of mouse melanoma cells, J. Supramol. Struct. 4:45.

    Article  CAS  Google Scholar 

  • Varga, J. M., Moellman, G., Fritsch, P., Godavska, E., and Lerner, A. B., 1976b, Association of cell surface receptors for melanotropin with the Golgi region in mouse melanoma cells, Proc. Natl. Acad. Sci. USA 73:559.

    Article  CAS  Google Scholar 

  • Vogel, Z., and Daniels, M. P., 1976, Ultrastructure of acetylcholine receptor clusters on cultured muscle fibers, J. Cell Biol. 69:501.

    Article  PubMed  CAS  Google Scholar 

  • Vogel, Z., and Nirenberg, M., 1976, Localization of acetylcholine receptors during synaptogenesis in retina, Proc. Natl. Acad. Sci. USA 73:1806.

    Article  PubMed  CAS  Google Scholar 

  • Vogel, Z., Malone, G. T., Ling, A., and Daniels, M. P., 1977, Identification of synaptic acetylcholine receptor sites in retina with peroxidase-labeled α-bungarotoxin, Proc. Natl. Acad. Sci. USA 74:3268.

    Article  PubMed  CAS  Google Scholar 

  • Weibel, E. R., Kistler, G. S., and Scherle, W. F., 1966, Practical stereological methods for morphometric cytology, J. Cell Biol. 30:23.

    Article  PubMed  CAS  Google Scholar 

  • Wernig, A. Stover, H., and Tonge, D., 1977, The labelling of motor endplates in skeletal muscle of mice with 125I-tetanus toxin, Arch. Pharmacol. 298:37.

    Article  CAS  Google Scholar 

  • Williams, L. T., and Lefkowitz, R. J., 1976, Alpha-adrenergic receptor identification by [3H]dihydro-ergocryptine binding, Science 192:791.

    Article  PubMed  CAS  Google Scholar 

  • Williams, M. A., 1973, Electron microscope autoradiography: its application to protein biosynthesis, in: Techniques in Protein Biosynthesis, Vol. 3 (P. N. Campbell and J. R. Sargent, eds.), pp. 126–190, Academic Press, New York.

    Google Scholar 

  • Yamamura, H. I., and Snyder, S. H., 1974, Muscarinic cholinergic receptor binding in the longitudinal muscle of the guinea pig ileum with [3H]quinuclidinyl benzilate, Mol. Pharmacol. 10:861.

    CAS  Google Scholar 

  • Young, A. B., and Snyder, S. H., 1974, Strychnine binding in rat spinal cord membranes associated with the synaptic glycine receptor: Co-operativity of glycine interactions, Mol. Pharmacol. 10:790.

    CAS  Google Scholar 

  • Zurn, A. D., and Fulpius, B. W., 1976, Accessibility to antibodies of acetylcholine receptors in the neuromuscular junction, Clin. Exp. Immunol. 24:9.

    PubMed  CAS  Google Scholar 

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Barnard, E.A. (1979). Visualization and Counting of Receptors at the Light and Electron Microscope Levels. In: O’Brien, R.D. (eds) General Principles and Procedures. The Receptors, vol 1. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-0979-6_7

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