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. 1991 Nov;104(3):603–608. doi: 10.1111/j.1476-5381.1991.tb12476.x

The change in the threshold for short-term desensitization in isolated smooth muscle cells showing an all-or-none response to acetylcholine.

M Mita 1, M K Uchida 1
PMCID: PMC1908226  PMID: 1797324

Abstract

1. Isolated smooth muscle cells from guinea-pig taenia caecum were prepared by collagenase digestion. The cells showed an all-or-none response to acetylcholine (ACh) under our experimental conditions. 2. Desensitized cells showed an all-or-none response but required a higher concentration of ACh for induction of contraction, indicating that the desensitization was due to a change in the threshold concentration. 3. In [3H]-quinuclidinyl benzilate ([3H]-QNB) binding to the desensitized cells, KD and Bmax were not significantly different from those estimated in the control cells. The competitive inhibition curve for specific binding of [3H]-QNB by ACh in the desensitized cells was in agreement with that of control cells. 4. The ACh-stimulated increase of the 45Ca2+ influx was very rapid and correlated well with the contraction of the cells. The concentrations of ACh inducing the maximal 45Ca2+ influx were increased by desensitization. 5. These results indicated that although the binding of ACh to the receptor was not changed by desensitization, the threshold concentration of ACh for their contraction was raised by desensitization, and the 45Ca2+ influx accompanying the contraction was shifted to the side of high concentration of ACh. 6. These results suggest that the development of short-term desensitization is due to an uncoupling of the receptor from the mechanism for initiation of the contraction.

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Selected References

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  1. Bagby R. M., Young A. M., Dotson R. S., Fisher B. A., McKinnon K. Contraction of single smooth muscle cells from Bufo marinus stomach. Nature. 1971 Dec 10;234(5328):351–352. doi: 10.1038/234351a0. [DOI] [PubMed] [Google Scholar]
  2. Benovic J. L., Strasser R. H., Caron M. G., Lefkowitz R. J. Beta-adrenergic receptor kinase: identification of a novel protein kinase that phosphorylates the agonist-occupied form of the receptor. Proc Natl Acad Sci U S A. 1986 May;83(9):2797–2801. doi: 10.1073/pnas.83.9.2797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bitar K. N., Bradford P., Putney J. W., Jr, Makhlouf G. M. Cytosolic calcium during contraction of isolated mammalian gastric muscle cells. Science. 1986 May 30;232(4754):1143–1145. doi: 10.1126/science.3704641. [DOI] [PubMed] [Google Scholar]
  4. Bitar K. N., Zfass A. M., Makhlouf G. M. Interaction of acetylcholine and cholecystokinin with dispersed smooth muscle cells. Am J Physiol. 1979 Aug;237(2):E172–E176. doi: 10.1152/ajpendo.1979.237.2.E172. [DOI] [PubMed] [Google Scholar]
  5. Bolton T. B., Clark J. P., Kitamura K., Lang R. J. Evidence that histamine and carbachol may open the same ion channels in longitudinal smooth muscle of guinea-pig ileum. J Physiol. 1981 Nov;320:363–379. doi: 10.1113/jphysiol.1981.sp013955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. El-Fakahany E., Richelson E. Involvement of calcium channels in short-term desensitization of muscarinic receptor-mediated cyclic GMP formation in mouse neuroblastoma cells. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6897–6901. doi: 10.1073/pnas.77.11.6897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fay F. S., Singer J. J. Characteristics of response of isolated smooth muscle cells to cholinergic drugs. Am J Physiol. 1977 Mar;232(3):C144–C154. doi: 10.1152/ajpcell.1977.232.3.C144. [DOI] [PubMed] [Google Scholar]
  8. Hasegawa M., Kobayashi M., Oyamada H., Kamishima T., Yoshida C., Ohata H., Maruyama I., Momose K., Gomi Y. Studies on isolated smooth muscle cells. IX. Application of papain for isolation of single smooth muscle cells from guinea-pig taenia coli. Nihon Heikatsukin Gakkai Zasshi. 1987 Feb;23(1):35–46. doi: 10.1540/jsmr1965.23.35. [DOI] [PubMed] [Google Scholar]
  9. Higuchi H., Takeyasu K., Uchida S., Yoshida H. Mechanism of agonist-induced degradation of muscarinic cholinergic receptor in cultured vas deferens of guinea-pig. Eur J Pharmacol. 1982 Apr 8;79(1-2):67–77. doi: 10.1016/0014-2999(82)90576-3. [DOI] [PubMed] [Google Scholar]
  10. Hishinuma S., Uchida M. K. Short-term desensitization of guinea-pig taenia caecum induced by carbachol occurs at intracellular Ca stores and that by histamine at H1-receptors. Br J Pharmacol. 1988 Jul;94(3):882–889. doi: 10.1111/j.1476-5381.1988.tb11600.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hishinuma S., Uchida M. K. Short-term desensitization of phosphatidylinositol turnover via muscarinic acetylcholine receptors and histamine H1-receptors in smooth muscle. Biochem Biophys Res Commun. 1989 Jul 31;162(2):733–739. doi: 10.1016/0006-291x(89)92371-1. [DOI] [PubMed] [Google Scholar]
  12. Ho A. K., Ling Q. L., Duffield R., Lam P. H., Wang J. H. Phosphorylation of brain muscarinic receptor: evidence of receptor regulation. Biochem Biophys Res Commun. 1987 Feb 13;142(3):911–918. doi: 10.1016/0006-291x(87)91500-2. [DOI] [PubMed] [Google Scholar]
  13. Horio S., Shima M., Ueda H., Ishida Y. Temperature-dependence of desensitization induced by acetylcholine and histamine in guinea-pig ileal longitudinal muscle. Br J Pharmacol. 1990 Jul;100(3):636–640. doi: 10.1111/j.1476-5381.1990.tb15859.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Huganir R. L., Delcour A. H., Greengard P., Hess G. P. Phosphorylation of the nicotinic acetylcholine receptor regulates its rate of desensitization. Nature. 1986 Jun 19;321(6072):774–776. doi: 10.1038/321774a0. [DOI] [PubMed] [Google Scholar]
  15. Irvine R. F. Inositol phosphates and calcium entry. 1987 Jul 30-Aug 5Nature. 328(6129):386–386. doi: 10.1038/328386b0. [DOI] [PubMed] [Google Scholar]
  16. Irvine R. F., Moor R. M. Micro-injection of inositol 1,3,4,5-tetrakisphosphate activates sea urchin eggs by a mechanism dependent on external Ca2+. Biochem J. 1986 Dec 15;240(3):917–920. doi: 10.1042/bj2400917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kagiya T., Uchida S., Mizushima A., Yoshida H. Alterations in muscarinic receptors of ventricular muscle in carbachol-induced short-term desensitization. Jpn J Pharmacol. 1986 Aug;41(4):511–517. doi: 10.1254/jjp.41.511. [DOI] [PubMed] [Google Scholar]
  18. Koike K., Ohtsuki H., Takayanagi I. Relationship between sensitivity and density of muscarinic receptors in single smooth muscle cells of guinea pig taenia caecum prepared under three conditions. Can J Physiol Pharmacol. 1990 Aug;68(8):1143–1147. doi: 10.1139/y90-171. [DOI] [PubMed] [Google Scholar]
  19. Kuriyama H., Tomita T. The action potential in the smooth muscle of the guinea pig taenia coli and ureter studied by the double sucrose-gap method. J Gen Physiol. 1970 Feb;55(2):147–162. doi: 10.1085/jgp.55.2.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kwatra M. M., Hosey M. M. Phosphorylation of the cardiac muscarinic receptor in intact chick heart and its regulation by a muscarinic agonist. J Biol Chem. 1986 Sep 25;261(27):12429–12432. [PubMed] [Google Scholar]
  21. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  22. Leeb-Lundberg L. M., Cotecchia S., Lomasney J. W., DeBernardis J. F., Lefkowitz R. J., Caron M. G. Phorbol esters promote alpha 1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5651–5655. doi: 10.1073/pnas.82.17.5651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Leurs R., Smit M. J., Bast A., Timmerman H. Different profiles of desensitization dynamics in guinea-pig jejunal longitudinal smooth muscle after stimulation with histamine and methacholine. Br J Pharmacol. 1990 Dec;101(4):881–888. doi: 10.1111/j.1476-5381.1990.tb14175.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Michell R. H. Inositol phospholipids and cell surface receptor function. Biochim Biophys Acta. 1975 Mar 25;415(1):81–47. doi: 10.1016/0304-4157(75)90017-9. [DOI] [PubMed] [Google Scholar]
  25. Mita M., Matsuo N., Uchida M. K. Effect of Ca2+ deprivation on short-term desensitization of isolated smooth muscle cells showing an all-or-none response to acetylcholine. Gen Pharmacol. 1988;19(3):441–445. doi: 10.1016/0306-3623(88)90044-4. [DOI] [PubMed] [Google Scholar]
  26. Mita M., Uchida M. K. Desensitization of isolated smooth muscle cells from guinea pig taenia caecum to acetylcholine. Can J Physiol Pharmacol. 1987 Mar;65(3):293–297. doi: 10.1139/y87-051. [DOI] [PubMed] [Google Scholar]
  27. Mita M., Uchida M. K. Muscarinic receptor binding and Ca2+ influx in the all-or-none response to acetylcholine of isolated smooth muscle cells. Eur J Pharmacol. 1988 Jun 22;151(1):9–17. doi: 10.1016/0014-2999(88)90686-3. [DOI] [PubMed] [Google Scholar]
  28. Momose K., Otsuka H., Higashikawa S., Gomi Y. Studies on isolated smooth muscle cells. VII. Response to agonists and the contraction velocity of taenia coli of guinea pig and the single smooth muscle cells. J Pharmacobiodyn. 1981 Sep;4(9):670–676. doi: 10.1248/bpb1978.4.670. [DOI] [PubMed] [Google Scholar]
  29. Momose K., Sato M., Gomi Y. Studies on isolated smooth muscle cells. VIII. 45Ca-efflux from single smooth muscle cells isolated from taenia coli of guinea pig. J Pharmacobiodyn. 1983 Jul;6(7):441–448. doi: 10.1248/bpb1978.6.441. [DOI] [PubMed] [Google Scholar]
  30. Nakazawa K., Matsuki N., Shigenobu K., Kasuya Y. Contractile response and electrophysiological properties in enzymatically dispersed smooth muscle cells of rat vas deferens. Pflugers Arch. 1987 Feb;408(2):112–119. doi: 10.1007/BF00581338. [DOI] [PubMed] [Google Scholar]
  31. Nasu T., Urakawa N. Effect of cholinergic drugs on calcium movement in guinea pig taenia coli. Jpn J Pharmacol. 1973 Aug;23(4):553–561. doi: 10.1254/jjp.23.553. [DOI] [PubMed] [Google Scholar]
  32. Scheid C. R., Fay F. S. Beta-adrenergic effects on transmembrane 45Ca fluxes in isolated smooth muscle cells. Am J Physiol. 1984 May;246(5 Pt 1):C431–C438. doi: 10.1152/ajpcell.1984.246.5.C431. [DOI] [PubMed] [Google Scholar]
  33. Seidel E. R., Johnson L. R. Contraction and [3H]QNB binding in collagenase isolated fundic smooth muscle cells. Am J Physiol. 1983 Aug;245(2):G270–G276. doi: 10.1152/ajpgi.1983.245.2.G270. [DOI] [PubMed] [Google Scholar]
  34. Sibley D. R., Lefkowitz R. J. Molecular mechanisms of receptor desensitization using the beta-adrenergic receptor-coupled adenylate cyclase system as a model. Nature. 1985 Sep 12;317(6033):124–129. doi: 10.1038/317124a0. [DOI] [PubMed] [Google Scholar]
  35. Siegel H., Jim K., Bolger G. T., Gengo P., Triggle D. J. Specific and non-specific desensitization of guinea-pig ileal smooth muscle. J Auton Pharmacol. 1984 Jun;4(2):109–125. doi: 10.1111/j.1474-8673.1984.tb00088.x. [DOI] [PubMed] [Google Scholar]
  36. Takayanagi I., Koike K., Hiruta T. Interactions of muscarinic drugs with their receptors in single cells of guinea-pig taenia caecum. J Pharm Pharmacol. 1986 Jun;38(6):476–478. doi: 10.1111/j.2042-7158.1986.tb04615.x. [DOI] [PubMed] [Google Scholar]
  37. Taylor J. E., El-Fakanany E., Richelson E. Long-term regulation of muscarinic acetylcholine receptors on cultured nerve cells. Life Sci. 1979 Dec 24;25(26):2181–2187. doi: 10.1016/0024-3205(79)90090-0. [DOI] [PubMed] [Google Scholar]
  38. Watson S. P., Stanley A. F., Sasaguri T. Does the hydrolysis of inositol phospholipids lead to the opening of voltage operated Ca2+ channels in guinea-pig ileum? Studies with fluoride ions and caffeine. Biochem Biophys Res Commun. 1988 May 31;153(1):14–20. doi: 10.1016/s0006-291x(88)81183-5. [DOI] [PubMed] [Google Scholar]
  39. Weller M., Morgan I. G. A possible role of the phosphorylation of synaptic membrane proteins in the control of calcium ion permeability. Biochim Biophys Acta. 1977 Mar 17;465(3):527–534. doi: 10.1016/0005-2736(77)90270-x. [DOI] [PubMed] [Google Scholar]

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