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. 1982;326:475–493. doi: 10.1113/jphysiol.1982.sp014207

Mechanisms of relaxation induced by activation of β-adrenoreceptors in smooth muscle cells of the guinea-pig mesenteric artery

Takeo Itoh 1, Hidetaka Izumi 1, Hirosi Kuriyama 1
PMCID: PMC1251489  PMID: 6286950

Abstract

Relaxation of smooth muscle cells induced by activation of β-adrenoceptors was investigated in intact and skinned muscles of the guinea-pig mesenteric artery.

1. In concentrations over 10-7 M, isoprenaline reduced the resting tone of intact preparations and also the amplitude of K contractions. When Ca was applied after previous superfusion with Ca-free solution, the amount of Ca accumulated into storage sites was increased by isoprenaline in polarized and depolarized ([K]o 128 mM) muscles. The amount of Ca stored increased even further when procaine and isoprenaline were applied simultaneously during store loading.

2. Isoprenaline increased the concentration of cyclic AMP as determined by radioimmunoassay. Application of isoprenaline at a concentration of 10-7 M increased cyclic AMP from 2.2±0.3 to 2.8±0.6 p-mole/mg wet weight and at 10-6 M increased it to 4.5±0.8 p-mole/mg wet weight after 5 min incubation (n = 4).

3. Application of cyclic AMP (3 × 10-6 M) with cyclic AMP-dependent protein kinase (50 μg/ml.) had no effect on the pCa—tension relationship in the skinned muscles. However, an increased concentration of cyclic AMP (> 10-5 M) suppressed the Ca-induced concentration only in the presence of protein kinase. This protein kinase (50 μg/ml.) alone had no effect on the Ca-induced contraction.

4. In skinned fibres, the Ca store could be loaded by applying low concentrations of Ca. If cyclic AMP (3 × 10-6 M) with protein kinase (50 μg/ml.) was applied during the loading procedure, the amount of Ca accumulated by the store increased if the loading solution contained 10-6 M-Ca applied for 2 min or less, but if the loading solution was applied for 3 min, or if higher Ca concentrations were used, the presence of cyclic AMP with protein kinase decreased the store size, suggesting that a Ca-induced Ca-release mechanism was also being activated.

5. In skinned muscles, accumulation of Ca into the store site in the presence of cyclic AMP (3 × 10-6 M) with protein kinase (50 μg/ml.) was further accelerated by simultaneous applications of procaine (5 mM), as here the Ca-induced Ca-release mechanism was suppressed.

6. These results indicate that activation of β-adrenoceptors by isoprenaline increases the amount of cyclic AMP in the intact muscles, and leads to an increase in Ca accumulation into the store site. In the skinned muscles, the Ca-induced Ca-release mechanism is activated by cyclic AMP and the Ca receptor for contraction (leiotonin C or calmodulin) is somewhat suppressed. These effects of exogenously applied cyclic AMP require the presence of protein kinase. The relaxation following β-adrenoceptor activation is more likely to involve Ca extrusion from the cell and accumulation of Ca in internal storage sites than suppression of the binding of calmodulin with the myosin light chain kinase.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adelstein R. S., Conti M. A., Hathaway D. R., Klee C. B. Phosphorylation of smooth muscle myosin light chain kinase by the catalytic subunit of adenosine 3': 5'-monophosphate-dependent protein kinase. J Biol Chem. 1978 Dec 10;253(23):8347–8350. [PubMed] [Google Scholar]
  2. Adelstein R. S., Eisenberg E. Regulation and kinetics of the actin-myosin-ATP interaction. Annu Rev Biochem. 1980;49:921–956. doi: 10.1146/annurev.bi.49.070180.004421. [DOI] [PubMed] [Google Scholar]
  3. Andersson R., Nilsson K., Wikberg J., Johansson S., Mohme-Lundholm E., Lundholm L. Cyclic nucleotides and the contraction of smooth muscle. Adv Cyclic Nucleotide Res. 1975;5:491–518. [PubMed] [Google Scholar]
  4. Bhalla R. C., Webb R. C., Singh D., Brock T. Role of cyclic AMP in rat aortic microsomal phosphorylation and calcium uptake. Am J Physiol. 1978 May;234(5):H508–H514. doi: 10.1152/ajpheart.1978.234.5.H508. [DOI] [PubMed] [Google Scholar]
  5. Brading A. F., Burnett M., Sneddon P. The effect of sodium removal on the contractile response of the guinea-pig taenia coli to carbachol. J Physiol. 1980 Sep;306:411–429. doi: 10.1113/jphysiol.1980.sp013404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bülbring E., den Hertog A. The action of isoprenaline on the smooth muscle of the guinea-pig taenia coli. J Physiol. 1980 Jul;304:277–296. doi: 10.1113/jphysiol.1980.sp013324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Casteels R., Raeymaekers L. The action of acetylcholine and catecholamines on an intracellular calcium store in the smooth muscle cells of the guinea-pig taenia coli. J Physiol. 1979 Sep;294:51–68. doi: 10.1113/jphysiol.1979.sp012914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cheung W. Y. Cyclic 3',5'-nucleotide phosphodiesterase. Demonstration of an activator. Biochem Biophys Res Commun. 1970 Feb 6;38(3):533–538. doi: 10.1016/0006-291x(70)90747-3. [DOI] [PubMed] [Google Scholar]
  9. Cheung W. Y. Cyclic 3',5'-nucleotide phosphodiesterase. Evidence for and properties of a protein activator. J Biol Chem. 1971 May 10;246(9):2859–2869. [PubMed] [Google Scholar]
  10. Conti M. A., Adelstein R. S. Phosphorylation by cyclic adenosine 3':5'-monophosphate-dependent protein kinase regulates myosin light chain kinase. Fed Proc. 1980 Apr;39(5):1569–1573. [PubMed] [Google Scholar]
  11. Diamond J. Role of cyclic nucleotides in control of smooth muscle contraction. Adv Cyclic Nucleotide Res. 1978;9:327–340. [PubMed] [Google Scholar]
  12. Endo M. Calcium release from the sarcoplasmic reticulum. Physiol Rev. 1977 Jan;57(1):71–108. doi: 10.1152/physrev.1977.57.1.71. [DOI] [PubMed] [Google Scholar]
  13. Fitzpatrick D. F., Szentivanyi A. Stimulation of calcium uptake into aortic microsomes by cyclic AMP and cyclic AMP-dependent protein kinase. Naunyn Schmiedebergs Arch Pharmacol. 1977 Jul;298(3):255–257. doi: 10.1007/BF00500896. [DOI] [PubMed] [Google Scholar]
  14. Goldberg N. D., Haddox M. K., Nicol S. E., Glass D. B., Sanford C. H., Kuehl F. A., Jr, Estensen R. Biologic regulation through opposing influences of cyclic GMP and cyclic AMP: the Yin Yang hypothesis. Adv Cyclic Nucleotide Res. 1975;5:307–330. [PubMed] [Google Scholar]
  15. Hidaka H., Yamaki T., Totsuka T., Asano M. Selective inhibitors of Ca2+-binding modulator of phosphodiesterase produce vascular relaxation and inhibit actin-myosin interaction. Mol Pharmacol. 1979 Jan;15(1):49–59. [PubMed] [Google Scholar]
  16. Hirata M., Kuriyama H. Does activation of cyclic AMP dependent phosphorylation induced by beta-adrenergic agent control the tone of vascular muscle? J Physiol. 1980 Oct;307:143–161. doi: 10.1113/jphysiol.1980.sp013428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Honma M., Satoh T., Takezawa J., Ui M. An ultrasensitive method for the simultaneous determination of cyclic AMP and cyclic GMP in small-volume samples from blood and tissue. Biochem Med. 1977 Dec;18(3):257–273. doi: 10.1016/0006-2944(77)90060-6. [DOI] [PubMed] [Google Scholar]
  18. Itoh T., Kajiwara M., Kitamura K., Kuriyama H. Roles of stored calcium on the mechanical response evoked in smooth muscle cells of the porcine coronary artery. J Physiol. 1982 Jan;322:107–125. doi: 10.1113/jphysiol.1982.sp014026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Itoh T., Kuriyama H., Suzuki H. Excitation--contraction coupling in smooth muscle cells of the guinea-pig mesenteric artery. J Physiol. 1981 Dec;321:513–535. doi: 10.1113/jphysiol.1981.sp014000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kerrick W. G., Hoar P. E., Cassidy P. S., Bolles L., Malencik D. A. Calcium-regulatory mechanisms. Functional classification using skinned fibers. J Gen Physiol. 1981 Feb;77(2):177–190. doi: 10.1085/jgp.77.2.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kerrick W. G., Hoar P. E. Inhibition of smooth muscle tension by cyclic AMP-dependent protein kinase. Nature. 1981 Jul 16;292(5820):253–255. doi: 10.1038/292253a0. [DOI] [PubMed] [Google Scholar]
  22. Kishikawa T. Alterations in the properties of the rat myometrium during gestation and post partum. Jpn J Physiol. 1981;31(4):515–536. doi: 10.2170/jjphysiol.31.515. [DOI] [PubMed] [Google Scholar]
  23. Krebs E. G., Beavo J. A. Phosphorylation-dephosphorylation of enzymes. Annu Rev Biochem. 1979;48:923–959. doi: 10.1146/annurev.bi.48.070179.004423. [DOI] [PubMed] [Google Scholar]
  24. Marshall J. M., Kroeger E. A. Adrenergic influences on uterine smooth muscle. Philos Trans R Soc Lond B Biol Sci. 1973 Mar 15;265(867):135–148. doi: 10.1098/rstb.1973.0016. [DOI] [PubMed] [Google Scholar]
  25. Marx J. L. Calmodulin: a protein for all seasons. Science. 1980 Apr 18;208(4441):274–276. doi: 10.1126/science.6102798. [DOI] [PubMed] [Google Scholar]
  26. Mikawa T., Nonomura Y., Hirata M., Ebashi S., Kakiuchi S. Involvement of an acidic protein in regulation of smooth muscle contraction by the tropomyosin-leiotonin system. J Biochem. 1978 Dec;84(6):1633–1636. doi: 10.1093/oxfordjournals.jbchem.a132290. [DOI] [PubMed] [Google Scholar]
  27. Mueller E., van Breemen C. Role of intracellular Ca2+ sequestration in beta-adrenergic relaxation of a smooth muscle. Nature. 1979 Oct 25;281(5733):682–683. doi: 10.1038/281682a0. [DOI] [PubMed] [Google Scholar]
  28. Noiman E. S. Phosphorylation of smooth muscle myosin light chains by cAMP-dependent protein kinase. J Biol Chem. 1980 Dec 10;255(23):11067–11070. [PubMed] [Google Scholar]
  29. Oashi H., Oga A., Saito K. Enhancement of Ca-contractions by catecholamines and temperature dependency in the depolarized teania coli of the guinea-pig. Jpn J Pharmacol. 1973 Aug;23(4):467–477. doi: 10.1254/jjp.23.467. [DOI] [PubMed] [Google Scholar]
  30. Oashi H., Takewaki T., Okada T. Calcium and the contractile effect of carbachol in the depolarized guinea-pig taenia caecum. Jpn J Pharmacol. 1974 Aug;24(4):601–611. doi: 10.1254/jjp.24.601. [DOI] [PubMed] [Google Scholar]
  31. Saida K., Nonomura Y. Characteristics of Ca2+- and Mg2+-induced tension development in chemically skinned smooth muscle fibers. J Gen Physiol. 1978 Jul;72(1):1–14. doi: 10.1085/jgp.72.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Scheid C. R., Honeyman T. W., Fay F. S. Mechanism of beta-adrenergic relaxation of smooth muscle. Nature. 1979 Jan 4;277(5691):32–36. doi: 10.1038/277032a0. [DOI] [PubMed] [Google Scholar]
  33. Sparrow M. P., Mrwa U., Hofmann F., Rüegg J. C. Calmodulin is essential for smooth muscle contraction. FEBS Lett. 1981 Mar 23;125(2):141–145. doi: 10.1016/0014-5793(81)80704-1. [DOI] [PubMed] [Google Scholar]
  34. Takata Y. Regional differences in electrical and mechanical properties of guinea-pig mesenteric vessels. Jpn J Physiol. 1980;30(5):709–728. doi: 10.2170/jjphysiol.30.709. [DOI] [PubMed] [Google Scholar]
  35. Thorens S., Haeusler G. Effects of adenosine 3' : 5'-monophosphate and guanosine 3' : 5'-monophosphate on calcium uptake and phosphorylation in membrane fractions of vascular smooth muscle. Biochim Biophys Acta. 1978 Sep 22;512(2):415–428. doi: 10.1016/0005-2736(78)90264-x. [DOI] [PubMed] [Google Scholar]
  36. Vallet B., Molla A., Demaille J. G. Cyclic adenosine 3',5'-monophosphate-dependent regulation of purified bovine aortic calcium/calmodulin-dependent myosin light chain kinase. Biochim Biophys Acta. 1981 May 5;674(2):256–264. doi: 10.1016/0304-4165(81)90383-4. [DOI] [PubMed] [Google Scholar]
  37. Waisman D. M., Stevens F. C., Wang J. H. Purification and characterization of a Ca2+-binding protein in Lumbricus terrestris. J Biol Chem. 1978 Feb 25;253(4):1106–1113. [PubMed] [Google Scholar]
  38. Webb R. C., Bhalla R. C. Calcium sequestration by subcellular fractions isolated from vascular smooth muscle: effect of cyclic nucleotides and prostaglandins. J Mol Cell Cardiol. 1976 Feb;8(2):145–157. doi: 10.1016/0022-2828(76)90026-2. [DOI] [PubMed] [Google Scholar]
  39. Yagi K., Yazawa M., Kakiuchi S., Ohshima M., Uenishi K. Identification of an activator protein for myosin light chain kinase as the Ca2+-dependent modulator protein. J Biol Chem. 1978 Mar 10;253(5):1338–1340. [PubMed] [Google Scholar]

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