Abstract
We have characterized and quantified the binding of [3H]CGP-12177 to β-adrenergic receptor sites in slices (300 microns) of rat cerebral cortex. The receptors are stereospecific, saturable and of high affinity. Binding of [3H]CGP is readily reversible and demonstrates appropriated drug specificty. This assay method allows the demonstration of isoproterenol-induced down-regulation (internalization) of β-adrenoreceptors. Receptor recycling is observed at 37°C in the absence of β-agonist but can be blocked by low temperature (0°C) or by monensin. β-Adrenoreceptors can also be labeled and quantified in intact, single pineal glands of rat, mouse and hamster. Rat pineals contain approximately 10 times more binding sites than do hamster or mouse pineals and up to 8 times more sites than found in rat cerebral cortex. Rat pineal [3H]CGP binding can be up- and down-regulated but not to the same degree as seen in brain slices. This assay method is simple, rapid and provides new opportunities for the study of other receptor types in intact tissue.
Similar content being viewed by others
References
Toews, M. L., andPerkins, J. P. 1984. Agonist-induced changes in β-adrenergic receptors on intact cells. J. Biol. Chem. 259:2227–2235.
Strulovici, B., andLefkowitz, R. J. 1984. Activation, desensitization, and recycling of frog erythrocyte β-adrenergic receptors. J. Biol. Chem. 259:4389–4395.
Hatton, G. I. 1983. The brain in slices: new approaches to old problems. Fed. Proc. 42:2863–2864.
Dingledine, R. (ed.) 1984. Brain Slices, Plenum Press, New York.
Bhanot, R., andWilkinson, M. 1983. Ontogeny of hypothalamic opiate ([3H]naloxone) binding sites assayed with a brain slice technique. Int. J. Develop. Neurosci. 1:165–170.
Whitaker, K. J., Manchester, E. L., Jacobson, W., andWilkinson, M. 1984. Benzodiazepine ([3H]flunitrazepam) binding sites in cerebellar and cerebral slices of mouse brain. Brain Res. Bull. 12:215–219.
Jacobson, W., andWilkinson, M. 1984. Opiate ([3H]naloxone) binding to hypothalamic and cerebral cortical slices of mouse brain. Brain Res. Bull. 13:481–485.
Staehelin, M., andHertel, C. 1983. [3H]CGP-12177, a β-adrenergic ligan suitable for measuring cell surface receptors. J. Recept. Res. 3:35–43.
Burt, D. R. 1978. Criteria for receptor identification. Pages 41–55, inYamamura, M. I., Enna, S. J., andKuhar, M. J. (eds.) Neurotransmitter receptor binding, Raven Press, New York.
Zivin, J. A., andWaud, D. R. 1982. How to analyze binding, enzyme and uptake data: the simplest case of a single phase. Life Sci. 30:1407–1422.
Basu, S. K., Goldstein, J. L., Anderson, R. G. W., andBrown, M. S. 1981. Monensin interrupts the recycling of low density lipoprotein receptors in human fibroblasts. Cell. 24:493–502.
Geary, W. A., andWooten, G. F. 1983. Quantitative film autoradiography of opiate agonist and antagonist binding in rat brain. J. Pharmacol. Exp. Therap. 225:234–240.
Chuang, D.-M., Kinnier, W. J., Farber, L., andCosta, E. 1980. A biochemical study of receptor internalization during β-adrenergic receptor desensitization in frog erythrocytes. Mol. Pharmacol. 18:348–355.
Wagner, H. R., andDavis, J. N. 1979. β-Adrenergic receptor regulation by agonists and membrane depolarization in rat brain slices. Proc. Nat. Acad. Sci. USA 76:2057–2061.
Dibner, M. D., andMolinoff, P. B. 1979. Agonist-induced changes in beta adrenergic receptor density and receptor-mediated responsiveness in slices of rat cerebral cortex. J. Pharmacol. Exp. Therap. 210:433–439.
Maggi, A., U'Prichard, D. C., andEnna, S. J. 1980. β-Adrenergic regulation of 838-1 receptors in the central nervous system. Science 207:645–657.
Hertel, C., andStaehelin, M. 1983. Reappearance of β-adrenergic receptors after isoproterenol treatment in intact C6 cells. J. Cell Biol. 97:1538–1543.
Kebabian, J. W., Zatz, M., Romero, J. A., andAxelrod, J. 1975. Rapid changes in rat pineal β-adrenergic receptor: Alterations in [3H]-alprenolol binding and adenylate cyclase. Proc. Nat. Acad. Sci. 72:3735–3739.
Craft, C. M., Morgan, W. W., Jones, D. J., andReiter, R. J. 1984. Hamster and rat pineal gland β-adrenoceptor characterization with iodocyanopindolol and the effect of decreased catecholamine synthesis on the receptor. Soc. Neurosci. Abstr. 10:819.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wilkinson, M., Wilkinson, D.A. Beta-adrenergic ([3H]CGP-12177) binding to brain slices and single intact pineal glands. Neurochem Res 10, 829–839 (1985). https://doi.org/10.1007/BF00964540
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00964540