Abstract
The regulation of a calcium-activated nonselective cation (Ca-NS+) channel by analogues of cyclic AMP has been investigated in the rat insulinoma cell line, CRI-G1. The activity of the channel is modulated by cyclic AMP in a complex way. In the majority of patches (83%) tested concentrations of cyclic AMP of 10 μm and above cause an inhibition of channel activity which is immediately reversible on washing. In contrast, lower concentrations of cyclic AMP, between 0.1 and 1.0 μm, produce a transient activation of channel activity in most patches (63%) tested. One group of analogues, including N6-monobutyryl cyclic AMP and N6, 2′-O-dibutyryl cyclic AMP reduced the activity of the Ca-NS+ channel at all concentrations tested and 2′-O-Monobutyryl cyclic AMP produced inhibition in all patches tested except one, at all concentrations. A second group produced dual concentration-dependent effects on Ca-NS+, low concentrations stimulating and high concentrations inhibiting channel activity. 6-Chloropurine cyclic AMP and 8-bromo cyclic AMP produced effects similar to those of cyclic AMP itself. In contrast, 8-[4-chlorophenylthio] cyclic AMP also showed a dual action, but with a high level of activation at all concentrations tested up to 1mm. Ca-NS+ channel activity was also predominantly activated by low concentrations of Sp-cAMPS. The activating effects of both Sp-cAMPS and cyclic AMP are antagonized by Rp-cAMPS, which by itself only produced a weak inhibition of Ca-NS+ channel activity even at concentrations of 10 μm and above. The results are discussed in terms of a model in which cyclic AMP, and other cyclic nucleotides, modulate the activity of the Ca-NS+ channel by binding to two separate sites.
Similar content being viewed by others
References
Ämmälä, C., Ashcroft, F.M., Rorsman, P. 1993. Calcium-independent potentiation of insulin release by cyclic AMP in single β-cells. Nature 363:356–358.
Ashcroft, F.M., Rorsman, P. 1991. Electrophysiology of the pancreaticβ-cell. Prog. Biophys. Molec. Biol. 54:87–143.
Carrington, C.A., Rubery, E.D., Pearson, E.C., Hales, C.N. 1986. Five new insulin-producing cell lines with differing secretory properties. J. Endocrinol. 109:193–200.
Corbin, J.D., Rannels, S.R., Flockhart, D.A., Robinson-Steiner, A.M., Tigani, M.C., Doskeland, S.O., Suva, R.H., Miller, J.P. 1982. Effect of cyclic nucleotide analogs on intrachain site 1 of protein kinase isozymes. Eur. J. Biochem. 125:259–266.
Dempster, J. 1988. Computer analysis of electrophysiological signals. In: Microcomputers in physiology: a practical approach. P.J. Fraser, Editor pp. 51–93. IRL, Oxford.
De Weille, J.R., Schmid-Antomarchi, H., Fosset, M., Lazdunski, M. 1989. Regulation of ATP-sensitive K+ channels in insulinoma cells: Activation by somatostatin and protein kinase C and the role of cAMP. Proc. Natl. Acad. Sci. USA 86:2971–2975.
DiFrancesco, D., Tortora, P. 1991. Direct activation of cardiac pacemaker channels by intracellular cyclic AMP. Nature 351:145–147.
Doskeland, S.O., Ogreid, D., Ekanger, R., Sturm, P.A., Miller, J.P., Suva, R.H. 1983. Mapping of the two intrachain cyclic nucleotide binding sites of adenosine cyclic 3′-5′-phosphate dependent protein kinase I. Biochemistry 22:1094–1101.
Dostmann, W.R.G., Taylor, S.S., Genieser, H.-G., Jastorff, B., Doskeland, S.O., Ogreid, D. 1990. Probing the cyclic nucleotide binding sites of cAMP-dependent protein kinases I and II with analogs of adenosine 3′,5′-cyclic phosphorothioates. J. Biol. Chem. 265:10484–10491.
Dryer, S.E., Henderson, D. 1991. A cyclic GMP-activated channel in dissociated cells of the chick pineal gland. Nature 353:756–758.
Dryer, S.E., Henderson, D. 1993. Cyclic GMP-activated channels of the chick pineal gland: effects of divalent cations, pH, and cyclic AMP. J. Comp. Physiol 172:271–279.
Fesenko, E., Kolesnikov, S., Lyubarsky, A. 1985. Induction by cyclic GMP of cationic conductance in plasma membrane of retinal rod outer segment. Nature 313:310–313.
Frings, S., Lynch, J.W., Lindemann, B. 1992. Properties of cyclic nucleotide-gated channels mediating olfactory transduction. J. Gen. Physiol. 100:45–67.
Hamill, O.P., Marty, A., Neher, E., Sakmann, B., Sigworth, F.J. 1981. Improved patch-clamp techniques for high resolution recording from cells and cell-free membrane patches. Pfleugers Arch. 391:85–100.
Haynes, L., Yau, K. 1985. Cyclic GMP-sensitive conductance in outer segment membrane of catfish cones. Nature 317:61–64.
Hockberger, P.E., Swandulla, D. 1987. Direct ion channel gating: a new function for intracellular messengers. Cell. Mol. Neurobiol. 7:229–236.
Ildefonse, M., Crouzy, S., Bennett, N. 1992. Gating of retinal rod cation channel by different nucleotides: comparative study of unitary currents. J. Membrane Biol. 130:91–104.
Kaupp, U.B. 1991. The cyclic nucleotide-gated channels of vertebrate photoreceptors and olfactory epithelium. Trends Neurosci. 14:150–157.
Kleene, S.J. 1993. The cyclic nucleotide-activated conductance in olfactory cilia: effects of cytoplasmic Mg2+ and Ca2+. J. Membrane Biol. 131:237–243.
Kozlowski, R.Z., Ashford, M.L.J. 1990. ATP-sensitive K+-channel run-down is Mg2+ dependent. Proc. R. Soc. Lond 240:397–410.
Kozlowski, R.Z., Hales, C.N., Ashford, M.L.J. 1989. Dual effects of diazoxide on ATP-K+-currents recorded from an insulin-secreting cell line. Br. J. Pharmacol. 97:1039–1050.
Kramer, R.H., Siegelbaum, S.A. 1992. Intracellular Ca2+ regulates the sensitivity of cyclic nucleotide-gated channels in olfactory receptor neurons. Neuron 9:897–906.
Nakamura, T., Gold, G. 1987. A cyclic nucleotide-gated conductance in olfactory receptor cilia. Nature 325:442–444.
Nizzari, M., Sesti, F., Giraudo, M.T., Virgino, C., Cattaneo, A., Torre, V. 1993. Single-channel properties of cloned cGMP-activated channels from retinal rods. Proc. R. Soc. Lond. 254:69–74.
Paulais, M., Teulon, J. 1989. A cation channel in the thick ascending limb of Henle's loop of the mouse kidney: inhibition by adenine nucleotides. J. Physiol 413:315–327.
Prentki, M., Matschinsky, F.M. 1987. Ca2+, cAMP, and phospholipidderived messengers in coupling mechanisms of insulin secretion. Physiol. Rev. 67:1185–1248.
Reale, V., Hales, C.N., Ashford, M.L.J. 1992. Cyclic AMP regulates a calcium-activated nonselective cation channel in a rat insulinoma cell line. J. Physiol. 446:312P.
Reale, V., Hales, C.N., Ashford, M.L.J. 1994a.. Nucleotide regulation of a calcium-activated cation channel in the rat insulinoma cell line, CRI-G1. J. Membrane Biol. 141:101–112.
Reale, V., Hales, C.N., Ashford, M.L.J. 1994b. The effects of pyridine nucleotides on the activity of a calcium-activated nonselective cation channel in the rat insulinoma cell line, CRI-G1. J. Membrane Biol. 142:299–307.
Rothermel, J.D., Perillo, N.L., Marks, J.S., Botelho, L.H.P. 1984. Effects of the specific cAMP antagonist, (Rp)-adenosine cyclic 3′,5′-phosphorothioate, on the cAMP-dependent protein kinase-induced activation of hepatic glycogen phosphorylase and glycogen synthase. J. Biol. Chem. 259:15294–15300.
Scholuebbers, H.G., Van Knippenberg, P.H., Baraniak, J., Stec, W.J., Morr, M., Jastorff, B. 1984. Investigations on stimulation of lac transcription in vivo in Escherichia coli by cAMP analogs. Eur. J. Biochem. 138:101–109.
Shabb, J.B., Corbin, J.D. 1992. Cyclic nucleotide-binding domains in proteins having diverse functions. J. Biol. Chem. 267:5723–5726.
Simon, L.N., Shuman, D.A., Robins, R.K. 1973. The chemistry and biological properties of nucleotides related to nucleoside 3′,5′-cyclic phosphates. Adv. Cyclic Nucleotide Res. 3:225–353.
Swandulla, D., Partridge, L.D. 1990. Nonspecific cation channels. In: Potassium channels. Structure, classification, function and therapeutical potential. N.S. Cook, Editor. pp. 167–180. Ellis Horwood, Chichester.
Tanaka, J.C., Eccleston, J.F., Furman, R.E. 1989. Photoreceptor channel activation by nucleotide derivatives. Biochemistry 28:2776–2784.
Van Haastert, P.J.M. 1983. Binding of cAMP and adenosine derivatives to Dictyostelium discoideum cells. J. Biol. Chem. 258:9643–9648.
Van Haastert, P.J.M., Kien, E. 1983. Binding of cAMP derivatives to Dictyostelium discoideum cells. Activation mechanism of the cell surface receptor. J. Biol. Chem. 258:9636–9642.
Weber, I.T., Steitz, T.A. 1987. Structure of a complex of catabolite gene activator protein and cyclic AMP refined at 2Å resolution. J. Mol. Biol. 198:311–326.
Weber, I.T., Steitz, T.A., Bubis, J., Taylor, S.S. 1987. Predicted structures of cAMP binding domains of type I and II regulatory subunits of cAMP-dependent protein kinase. Biochemistry 26:343–351.
Yau, K.-W. 1994. Cyclic nucleotide-gated channels: An expanding new family of ion channels. Proc. Natl. Acad. Sci. USA 91:3481–3483.
Zawalich, W.S., Rasmussen, H. 1990. Control of insulin secretion: a model involving Ca2+, cAMP and diacylglycerol. Mol. Cell. Endocrinol. 70:119–137.
Zawalich, W.S., Zawalich, K.C. 1990. Forskolin-induced desensitization of pancreatic β-cell insulin secretory responsiveness: Possible involvement of impaired information flow in the inositol-lipid cycle. Endocrinology 126:2307–2312.
Zimmerman, A.L., Yamanaka, G., Eckstein, F., Baylor, D.B., Stryer, L. 1985. Interactions of hydrolysis-resistant analogs of cyclic GMP with the phosphodiesterase and light-sensitive channel of retinal rod outer segments. Proc. Natl. Acad. Sci. USA 82:8813–8817.
Zufall, F., Firestein, S. 1993. Divalent cations block the cyclic nucleotide-gated channel of olfactory receptor neurons. J. Neurophysiol. 69:1758–1768.
Zufall, F., Hatt, H. 1991. Dual activation of a sex pheromonedependent ion channel from insect olfactory dendrites by protein kinase C activators and cyclic GMP. Proc. Natl. Acad. Sci. USA 88:8520–8524.
Zufall, F., Hatt, H., Keil, T.A. 1991. A calcium-activated nonspecific cation channel from olfactory receptor neurones of the silkmoth Antheraea polyphemus. J. Exp. Biol. 161:455–468.
Author information
Authors and Affiliations
Additional information
We thank Dr. J. Corbin for advice on cyclic nucleotide analogues, Dr. P.D. Evans for his comments on the manuscript and the British Diabetic Association, Serono Diagnostic (UK) Ltd., and the Wolfson Trust for support. V.R. was supported by a grant from Serono Diagnostics (UK) Ltd.
Rights and permissions
About this article
Cite this article
Reale, V., Hales, C.N. & Ashford, M.L.J. Regulation of calcium-activated nonselective cation channel activity by cyclic nucleotides in the rat insulinoma cell line, CRI-G1. J. Membarin Biol. 145, 267–278 (1995). https://doi.org/10.1007/BF00232718
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00232718