Abstract
1. Relationships between cytosolic Ca2+ concentration ([Ca2+]cyt), myosin light chain (MLC) phosphorylation and muscle tension were examined in circular smooth muscle of canine gastric antrum. 2. Electrical slow waves induced a transient increase in [Ca2+]cyt and muscle tension. [Ca2+]cyt increased before the initiation of contraction and reached a maximum before the peak of the phasic contractions. Following the first Ca2+ transient, a second rise in [Ca2+]cyt was often observed. The second Ca2+ transient was of similar magnitude to the first, but only in some cases was this increase in [Ca2+]cyt associated with a second phase of contraction. Relaxation occurred more rapidly than the restoration of resting levels of [Ca2+]cyt. 3. Acetylcholine (ACh; 3 x 10(-7) M) increased the amplitude of Ca2+ transients, caused MLC phosphorylation and increased the force of contraction. The decay of contraction and MLC dephosphorylation preceded that of [Ca2+]cyt. 4. Increasing external K+ (to 25-40 mM) caused a sustained increase in [Ca2+]cyt, but little change in resting tension. This suggests that the Ca2+ sensitivity decreased as [Ca2+]cyt increased. Increasing K+ to 59.5 mM further increased the level of [Ca2+]cyt, induced MLC phosphorylation and caused a transient contraction. When normal levels of K+ were restored, the rates of MLC dephosphorylation and relaxation exceeded the rate of decay in [Ca2+]cyt. 5. Removal of external Ca2+ in depolarized muscles decreased [Ca2+]cyt below the resting level without affecting resting tension. Readmission of Ca2+ to depolarized muscles caused force to develop at [Ca2+]cyt levels below the original resting level, suggesting that Ca2+ sensitivity was increased when the resting level of [Ca2+]cyt was decreased. 6. The phosphatase inhibitor, calyculin-A (10(-6) M), induced tonic contraction and MLC phosphorylation without an increase in [Ca2+]cyt. During these contractures, electrical activity caused transient increases in [Ca2+]cyt and phasic contractions which were superimposed upon the Ca(2+)-independent contracture. In the presence of calyculin-A, relaxation occurred in two phases. The initial, rapid phase of relaxation was not significantly affected by calyculin-A, but the slow phase was significantly decreased. 7. These results suggest that the relationship between [Ca2+]cyt, MLC phosphorylation and contraction changes as a function of [Ca2+]cyt in canine antral muscles. This may be due to a Ca(2+)-and time-dependent phosphatase that regulates the level of myosin phosphorylation.
Full text
PDFSelected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bauer A. J., Publicover N. G., Sanders K. M. Origin and spread of slow waves in canine gastric antral circular muscle. Am J Physiol. 1985 Dec;249(6 Pt 1):G800–G806. doi: 10.1152/ajpgi.1985.249.6.G800. [DOI] [PubMed] [Google Scholar]
- Bauer A. J., Reed J. B., Sanders K. M. Slow wave heterogeneity within the circular muscle of the canine gastric antrum. J Physiol. 1985 Sep;366:221–232. doi: 10.1113/jphysiol.1985.sp015793. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bauer A. J., Sanders K. M. Gradient in excitation-contraction coupling in canine gastric antral circular muscle. J Physiol. 1985 Dec;369:283–294. doi: 10.1113/jphysiol.1985.sp015901. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bialojan C., Takai A. Inhibitory effect of a marine-sponge toxin, okadaic acid, on protein phosphatases. Specificity and kinetics. Biochem J. 1988 Nov 15;256(1):283–290. doi: 10.1042/bj2560283. [DOI] [PMC free article] [PubMed] [Google Scholar]
- DeFeo T. T., Morgan K. G. Calcium-force relationships as detected with aequorin in two different vascular smooth muscles of the ferret. J Physiol. 1985 Dec;369:269–282. doi: 10.1113/jphysiol.1985.sp015900. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gerthoffer W. T., Murphey K. A., Gunst S. J. Aequorin luminescence, myosin phosphorylation, and active stress in tracheal smooth muscle. Am J Physiol. 1989 Dec;257(6 Pt 1):C1062–C1068. doi: 10.1152/ajpcell.1989.257.6.C1062. [DOI] [PubMed] [Google Scholar]
- Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
- Hathaway D. R., Haeberle J. R. A radioimmunoblotting method for measuring myosin light chain phosphorylation levels in smooth muscle. Am J Physiol. 1985 Sep;249(3 Pt 1):C345–C351. doi: 10.1152/ajpcell.1985.249.3.C345. [DOI] [PubMed] [Google Scholar]
- Hathaway D. R., Haeberle J. R. Selective purification of the 20,000-Da light chains of smooth muscle myosin. Anal Biochem. 1983 Nov;135(1):37–43. doi: 10.1016/0003-2697(83)90726-1. [DOI] [PubMed] [Google Scholar]
- Himpens B., Casteels R. Different effects of depolarization and muscarinic stimulation on the Ca2+/force relationship during the contraction-relaxation cycle in the guinea pig ileum. Pflugers Arch. 1990 Apr;416(1-2):28–35. doi: 10.1007/BF00370218. [DOI] [PubMed] [Google Scholar]
- Himpens B., Matthijs G., Somlyo A. P. Desensitization to cytoplasmic Ca2+ and Ca2+ sensitivities of guinea-pig ileum and rabbit pulmonary artery smooth muscle. J Physiol. 1989 Jun;413:489–503. doi: 10.1113/jphysiol.1989.sp017665. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ingebritsen T. S., Cohen P. The protein phosphatases involved in cellular regulation. 1. Classification and substrate specificities. Eur J Biochem. 1983 May 2;132(2):255–261. doi: 10.1111/j.1432-1033.1983.tb07357.x. [DOI] [PubMed] [Google Scholar]
- Ishihara H., Ozaki H., Sato K., Hori M., Karaki H., Watabe S., Kato Y., Fusetani N., Hashimoto K., Uemura D. Calcium-independent activation of contractile apparatus in smooth muscle by calyculin-A. J Pharmacol Exp Ther. 1989 Jul;250(1):388–396. [PubMed] [Google Scholar]
- Ito Y., Kuriyama H., Parker I. Calcium transients evoked by electrical stimulation of smooth muscle from guinea-pig ileum recorded by the use of Fura-2. J Physiol. 1988 Dec;407:117–134. doi: 10.1113/jphysiol.1988.sp017406. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kamm K. E., Stull J. T. The function of myosin and myosin light chain kinase phosphorylation in smooth muscle. Annu Rev Pharmacol Toxicol. 1985;25:593–620. doi: 10.1146/annurev.pa.25.040185.003113. [DOI] [PubMed] [Google Scholar]
- Karaki H. Ca2+ localization and sensitivity in vascular smooth muscle. Trends Pharmacol Sci. 1989 Aug;10(8):320–325. doi: 10.1016/0165-6147(89)90066-7. [DOI] [PubMed] [Google Scholar]
- Kitazawa T., Somlyo A. P. Desensitization and muscarinic re-sensitization of force and myosin light chain phosphorylation to cytoplasmic Ca2+ in smooth muscle. Biochem Biophys Res Commun. 1990 Nov 15;172(3):1291–1297. doi: 10.1016/0006-291x(90)91589-k. [DOI] [PubMed] [Google Scholar]
- Konishi M., Olson A., Hollingworth S., Baylor S. M. Myoplasmic binding of fura-2 investigated by steady-state fluorescence and absorbance measurements. Biophys J. 1988 Dec;54(6):1089–1104. doi: 10.1016/S0006-3495(88)83045-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuriyama H., Osa T., Tasaki H. Electrophysiological studies of the antrum muscle fibers of the guinea pig stomach. J Gen Physiol. 1970 Jan;55(1):48–62. doi: 10.1085/jgp.55.1.48. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mitsui M., Karaki H. Dual effects of carbachol on cytosolic Ca2+ and contraction in intestinal smooth muscle. Am J Physiol. 1990 May;258(5 Pt 1):C787–C793. doi: 10.1152/ajpcell.1990.258.5.C787. [DOI] [PubMed] [Google Scholar]
- Obara K., Takai A., Ruegg J. C., de Lanerolle P. Okadaic acid, a phosphatase inhibitor, produces a Ca2+ and calmodulin-independent contraction of smooth muscle. Pflugers Arch. 1989 Jun;414(2):134–138. doi: 10.1007/BF00580954. [DOI] [PubMed] [Google Scholar]
- Ozaki H., Ishihara H., Kohama K., Nonomura Y., Shibata S., Karaki H. Calcium-independent phosphorylation of smooth muscle myosin light chain by okadaic acid isolated from black sponge (Halichondria okadai). J Pharmacol Exp Ther. 1987 Dec;243(3):1167–1173. [PubMed] [Google Scholar]
- Ozaki H., Kohama K., Nonomura Y., Shibata S., Karaki H. Direct activation by okadaic acid of the contractile elements in the smooth muscle of guinea-pig taenia coli. Naunyn Schmiedebergs Arch Pharmacol. 1987 Mar;335(3):356–358. doi: 10.1007/BF00172811. [DOI] [PubMed] [Google Scholar]
- Ozaki H., Kwon S. C., Tajimi M., Karaki H. Changes in cytosolic CA2+ and contraction induced by various stimulants and relaxants in canine tracheal smooth muscle. Pflugers Arch. 1990 Jun;416(4):351–359. doi: 10.1007/BF00370740. [DOI] [PubMed] [Google Scholar]
- Ozaki H., Ohyama T., Sato K., Karaki H. Ca2(+)-dependent and independent mechanisms of sustained contraction in vascular smooth muscle of rat aorta. Jpn J Pharmacol. 1990 Mar;52(3):509–512. doi: 10.1254/jjp.52.509. [DOI] [PubMed] [Google Scholar]
- Ozaki H., Sato K., Satoh T., Karaki H. Simultaneous recordings of calcium signals and mechanical activity using fluorescent dye fura 2 in isolated strips of vascular smooth muscle. Jpn J Pharmacol. 1987 Nov;45(3):429–433. doi: 10.1254/jjp.45.429. [DOI] [PubMed] [Google Scholar]
- Ozaki H., Satoh T., Karaki H., Ishida Y. Regulation of metabolism and contraction by cytoplasmic calcium in the intestinal smooth muscle. J Biol Chem. 1988 Oct 5;263(28):14074–14079. [PubMed] [Google Scholar]
- Papasova M. P., Nagai T., Prosser C. L. Two-component slow waves in smooth muscle of cat stomach. Am J Physiol. 1968 Apr;214(4):695–702. doi: 10.1152/ajplegacy.1968.214.4.695. [DOI] [PubMed] [Google Scholar]
- Persechini A., Kamm K. E., Stull J. T. Different phosphorylated forms of myosin in contracting tracheal smooth muscle. J Biol Chem. 1986 May 15;261(14):6293–6299. [PubMed] [Google Scholar]
- Rembold C. M. Desensitization of swine arterial smooth muscle to transplasmalemmal Ca2+ influx. J Physiol. 1989 Sep;416:273–290. doi: 10.1113/jphysiol.1989.sp017760. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sakata K., Ozaki H., Kwon S. C., Karaki H. Effects of endothelin on the mechanical activity and cytosolic calcium level of various types of smooth muscle. Br J Pharmacol. 1989 Oct;98(2):483–492. doi: 10.1111/j.1476-5381.1989.tb12621.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sato K., Ozaki H., Karaki H. Changes in cytosolic calcium level in vascular smooth muscle strip measured simultaneously with contraction using fluorescent calcium indicator fura 2. J Pharmacol Exp Ther. 1988 Jul;246(1):294–300. [PubMed] [Google Scholar]
- Scanlon M., Williams D. A., Fay F. S. A Ca2+-insensitive form of fura-2 associated with polymorphonuclear leukocytes. Assessment and accurate Ca2+ measurement. J Biol Chem. 1987 May 5;262(13):6308–6312. [PubMed] [Google Scholar]
- Shibata S., Ishida Y., Kitano H., Ohizumi Y., Habon J., Tsukitani Y., Kikuchi H. Contractile effects of okadaic acid, a novel ionophore-like substance from black sponge, on isolated smooth muscles under the condition of Ca deficiency. J Pharmacol Exp Ther. 1982 Oct;223(1):135–143. [PubMed] [Google Scholar]
- Somlyo A. P., Himpens B. Cell calcium and its regulation in smooth muscle. FASEB J. 1989 Sep;3(11):2266–2276. doi: 10.1096/fasebj.3.11.2506092. [DOI] [PubMed] [Google Scholar]
- Somlyo A. P., Somlyo A. V. Flash photolysis studies of excitation-contraction coupling, regulation, and contraction in smooth muscle. Annu Rev Physiol. 1990;52:857–874. doi: 10.1146/annurev.ph.52.030190.004233. [DOI] [PubMed] [Google Scholar]
- Stewart A. A., Ingebritsen T. S., Cohen P. The protein phosphatases involved in cellular regulation. 5. Purification and properties of a Ca2+/calmodulin-dependent protein phosphatase (2B) from rabbit skeletal muscle. Eur J Biochem. 1983 May 2;132(2):289–295. doi: 10.1111/j.1432-1033.1983.tb07361.x. [DOI] [PubMed] [Google Scholar]
- Yagi S., Becker P. L., Fay F. S. Relationship between force and Ca2+ concentration in smooth muscle as revealed by measurements on single cells. Proc Natl Acad Sci U S A. 1988 Jun;85(11):4109–4113. doi: 10.1073/pnas.85.11.4109. [DOI] [PMC free article] [PubMed] [Google Scholar]
- el-Sharkawy T. Y., Morgan K. G., Szurszewski J. H. Intracellular electrical activity of canine and human gastric smooth muscle. J Physiol. 1978 Jun;279:291–307. doi: 10.1113/jphysiol.1978.sp012345. [DOI] [PMC free article] [PubMed] [Google Scholar]