Role of T-type $Ca^{2+}$ Channels in the Spontaneous Phasic Contraction of Pregnant Rat Uterine Smooth Muscle

  • Lee, Si-Eun (Department of Physiology, BK 21 Project for Medical Sciences, College of Medicine, Yonsei University) ;
  • Ahn, Duck-Sun (Department of Physiology, BK 21 Project for Medical Sciences, College of Medicine, Yonsei University) ;
  • Lee, Young-Ho (Department of Physiology, BK 21 Project for Medical Sciences, College of Medicine, Yonsei University)
  • Published : 2009.06.30

Abstract

Although extracellular $Ca^{2+}$ entry through the voltage-dependent $Ca^{2+}$ channels plays an important role in the spontaneous phasic contractions of the pregnant rat myometrium, the role of the T-type $Ca^{2+}$ channels has yet to be fully identified. The aim of this study was to investigate the role of the T-type $Ca^{2+}$ channel in the spontaneous phasic contractions of the rat myometrium. Spontaneous phasic contractions and $[Ca^{2+}]_i$ were measured simultaneously in the longitudinal strips of female Sprague-Dawley rats late in their pregnancy (on day 18 ${\sim}$ 20 of gestation: term=22 days). The expression of T-type $Ca^{2+}$ channel mRNAs or protein levels was measured. Cumulative addition of low concentrations (< 1 ${\mu}M$) of nifedipine, a L-type $Ca^{2+}$ channel blocker, produced a decrease in the amplitude of the spontaneous $Ca^{2+}$ transients and contractions with no significant change in frequency. The mRNAs and proteins encoding two subunits (${\alpha}$ 1G, ${\alpha}$ 1H) of the T-type $Ca^{2+}$ channels were expressed in longitudinal muscle layer of rat myometrium. Cumulative addition of mibefradil, NNC 55-0396 or nickel induced a concentration-dependent inhibition of the amplitude and frequency of the spontaneous $Ca^{2+}$ transients and contractions. Mibefradil, NNC 55-0396 or nickel also attenuated the slope of rising phase of spontaneous $Ca^{2+}$ transients consistent with the reduction of the frequency. It is concluded that T-type $Ca^{2+}$ channels are expressed in the pregnant rat myometrium and may play a key role for the regulation of the frequency of spontaneous phasic contractions.

Keywords

References

  1. Asokan KT, Sarkar SN, Mishra SK, Raviprakash V. Effects of mibefradil on uterine contractility. Eur J Pharmacol 455: 65−71, 2002 https://doi.org/10.1016/S0014-2999(02)02487-1
  2. Billman GE. Ro 40-5967, a novel calcium channel antagonist, protects against ventricular fibrillation. Eur J Pharmacol 229: 179−187, 1992 https://doi.org/10.1016/0014-2999(92)90553-G
  3. Blanks AM, Zhao ZH, Shmygol A, Bru-Mercier G, Astle S, Thornton S. Characterization of the molecular and electrophysiological properties of the T-type calcium channel in human myometrium. J Physiol 581(Pt 3): 915−926, 2007 https://doi.org/10.1113/jphysiol.2007.132126
  4. Challis JRG, Matthews SG, Gibb W, Lye SJ. Endocrine and paracrine regulation of birth at term and preterm. Endocrine Rev 21: 514−550, 2000 https://doi.org/10.1210/er.21.5.514
  5. Chien EK, Saunders T, Phillippe M. The mechanisms underlying Bay K 8644-stimulated phasic myometrial contractions. J Soc Gynecol Invest 3: 106−112, 1996 https://doi.org/10.1016/1071-5576(96)00006-8
  6. Chow KY, Wu C, Sui GP, Fry CH. Role of the T-type $Ca^{2+}$ current on the contractile performance of guinea pig detrusor smooth muscle. Neurourol Urodyn 22: 77−82, 2003 https://doi.org/10.1002/nau.10081
  7. Coleman HA, Hart JDE, Tonta MA, Parkington HC. Changes in the mechanisms involved in uterine contractions during pregnancy in guinea-pigs. J Physiol 523: 785−798, 2000 https://doi.org/10.1111/j.1469-7793.2000.00785.x
  8. Coleman HA, Parkington HC. The role of membrane potential in the control of uterine motility. In: Carsten ME, Miller JD ed, Uterine function: Molecular and cellular aspects. New York: Plenum Press, p 195−248, 1990
  9. Collins PL, Moore JJ, Idriss E, Kulp TM. Human fetal membranes inhibit calcium L-channel activated uterine contractions. Collins PL, Moore JJ, Idriss E, Kulp TM. Human fetal membranes inhibit calcium L-channel activated uterine contractions. Am J Obstet Gynecol 175: 1173−1179, 1996.175: 1173−1179, 1996 https://doi.org/10.1016/S0002-9378(96)70024-8
  10. Doerr T, Denger R, Trautwein W. Calcium currents in single SA nodal cells of the rabbit heart studied with action potential clamp. Pflugers Arch 413: 599−603, 1989 https://doi.org/10.1007/BF00581808
  11. Honore E, Amedee T, Martin C, Dacquet C, Mironneau C, Mironneau J. Calcium channel current and its sensitivity to (+) isradipine in cultured pregnant rat myometrial cells. Pflugers Arch 414: 477−483, 1989 https://doi.org/10.1007/BF00585060
  12. Huang L, Keyser BM, Tagmose TM, Hansen JB, Taylor JT, Zhuang H, Zhang M, Ragsdale DS, Li M. NNC 55-0396[(1S,2S)-2-(2- (N-[(3-benzimidazol-2-yl)propyl]-N-methylamino)ethyl)-6- fluoro-1,2,3,4-tetrahydro-1-isopropyl-2-naphytyl cyclopropanecarboxylate dihydrochloride]: a new selective inhibitor of T-type calcium channels. J Pharmacol Exp Therap 309: 193−199, 2004 https://doi.org/10.1124/jpet.103.060814
  13. noue Y, Sperelakis N. Gestational change in $Na^+$+ and $Ca^{2+}$ current densities in rat myometrial smooth muscle cells. Am J Physiol 260: C658−C663, 1991
  14. mari K, Mironneau C, Mironneau J. Inactivation of calcium channels current in rat uterine smooth muscle: evidence for calcium and voltage-mediated mechanisms. J Physiol 380: 111−126, 1986
  15. Knock GA, Aaronson PI. Calcium antagonistic properties of the cyclooxygenase-2 inhibitor nimesulide in human myometrial myocytes. Br J Pharmacol 127: 1470−1478, 1999 https://doi.org/10.1038/sj.bjp.0702685
  16. Lee JH, Gomora JC, Cribbs LL, Perez-Reyes E. Nickel block of three cloned T-type calcium channels: low concentrations selectively block $\alpha$1H. Biophys J 77: 3034−3042, 1999 https://doi.org/10.1016/S0006-3495(99)77134-1
  17. Mershon JL, Mikala G, Schwartz A. Changes in the expression of the L-type voltage-dependent calcium channel during pregnancy and parturition in the rat. Biol Reprod 51: 993−999, 1994 https://doi.org/10.1095/biolreprod51.5.993
  18. Mironneau J. Excitation-contraction coupling in voltage clamped uterine smooth muscle. J Physiol 233: 127−141, 1973
  19. Mishra SK, Hermsmeyer K. Selective inhibition of T-type $Ca^{2+}$ channels by Ro 40-5967. Circ Res 75: 144−148, 1994 https://doi.org/10.1161/01.RES.75.1.144
  20. Ohkubo T, Kawarabayashi T, Inoue Y, Kitamura K. Differential expression of L-and T-type calcium channels between longitudinal and circular muscles of the rat myometrium during pregnancy. Gynecol Obstet Invest 59: 80−85, 2005 https://doi.org/10.1159/000082333
  21. Ohya Y, Sperelakis N. Fast $Na^+$ and slow $Ca^{2+}$ channels in single uterine muscle cells from pregnant rats. Am J Physiol 257: C408−C412, 1989
  22. Parkington HC, Coleman HA. Ionic mechanisms underlying action potentials in myometrium. Clin Exp Pharmacol Physiol 15: 657−665, 1988 https://doi.org/10.1111/j.1440-1681.1988.tb01125.x
  23. Parkington HC, Coleman HA. Excitability in uterine smooth muscle. Front Horm Res 27: 179−200, 2001 https://doi.org/10.1159/000061026
  24. Parkington HC, Tonta MA, Brennecke SP, Coleman HA. Contractile activity, membrane potential, and cytoplasmic calcium in human uterine smooth muscle in the third trimester of pregnancy and during labor. Am J Obstet Gynecol 181: 1145−1151, 1999
  25. Perez-Reyes E. Molecular physiology of low-voltage-activated T-type calcium channels. Physiol Rev 83: 117−161, 2003
  26. Riemer RK, Heymann MA. Regulation of uterine smooth muscle function during gestation. Pediatrics Res 44: 615−627, 1998
  27. Shmigol AV, Eisner DA, Wray S. Properties of voltage-activated [$Ca^{2+}$]i transients in single smooth muscle cells isolated from pregnant rat uterus. J Physiol 511: 803−811, 1998 https://doi.org/10.1111/j.1469-7793.1998.803bg.x
  28. Sui GP, Wu C, Fry CH. Inward calcium currents in cultured and freshly isolated detrusor muscle cells: evidence of a T-type calcium current. J Urol 165: 621−626, 2001 https://doi.org/10.1097/00005392-200102000-00084
  29. Taggart MJ, Tribe RM. Cellular ionic mechanisms controlling uterine smooth muscle contraction: effects of gestational state. In: Savineau JP ed, New Frontiers in Smooth Muscle Biology and Physiology. India: Research Signpost, p 523−549, 2007
  30. Tytgat J, Vereecke J, Carmeliet E. Combined study of sodium current and T-type calcium current in isolated cardiac cells. Pflugers Arch 417: 142−148, 1990 https://doi.org/10.1007/BF00370691
  31. Wray S, Jones K, Kupittayanant S, Li Y, Matthew A, Monir-Bishty E, Noble K, Pierce SJ, Quenby S, Shmygol AV. Calcium signaling and uterine contractility. J Soc Gynecol Invest 10: 252−264, 2003 https://doi.org/10.1016/S1071-5576(03)00089-3
  32. Yeon DS, Kim JS, Ahn DS, Kwon SC, Kang BS, Morgan KG, Lee YH. Role of protein kinase C- or RhoA-induced $Ca^{2+}$ sensitization in stretch-induced myogenic tone. Cardiovas Res 53: 431−438, 2002 https://doi.org/10.1016/S0008-6363(01)00496-5
  33. Young RC, Smith LH, McLaren MD. T-type and L-type calcium currents in freshly dispersed human uterine smooth muscle cells. Am J Obstet Gynecol 169: 785−792, 2003