Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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TRPC4 Is an Essential Component of the Nonselective Cation Channel Activated by Muscarinic Stimulation in Mouse Visceral Smooth Muscle Cells

  • Lee, Kyu Pil (Department of Physiology and Biophysics, Seoul National University College of Medicine) ;
  • Jun, Jae Yeoul (Department of Physiology, College of Medicine, Chosun University) ;
  • Chang, In-Youb (Department of Anatomy, College of Medicine, Chosun University) ;
  • Suh, Suk-Hyo (Department of Physiology, College of Medicine, Ewha Womans University) ;
  • So, Insuk (Department of Physiology and Biophysics, Seoul National University College of Medicine) ;
  • Kim, Ki Whan (Department of Physiology and Biophysics, Seoul National University College of Medicine)
  • Received : 2005.08.30
  • Accepted : 2005.09.28
  • Published : 2005.12.31
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Abstract

Classical transient receptor potential channels (TRPCs) are thought to be candidates for the nonselective cation channels (NSCCs) involved in pacemaker activity and its neuromodulation in murine stomach smooth muscle. We aimed to determine the role of TRPC4 in the formation of NSCCs and in the generation of slow waves. At a holding potential of -60 mV, $50{\mu}M$ carbachol (CCh) induced $I_{NSCC}$ of amplitude [$500.8{\pm}161.8pA$ (n = 8)] at -60 mV in mouse gastric smooth muscle cells. We investigated the effects of commercially available antibodies to TRPC4 on recombinant TRPC4 expressed in HEK cells and CCh-induced NSCCs in gastric smooth muscle cells. TRPC4 currents in HEK cells were reduced from $1525.6{\pm}414.4pA$ (n = 8) to $146.4{\pm}83.3pA$ (n = 10) by anti-TRPC4 antibody and $I_{NSCC}$ amplitudes were reduced from $230.9{\pm}36.3pA$ (n = 15) to $49.8{\pm}11.8pA$ (n = 9). Furthermore, $I_{NSCC}$ in the gastric smooth muscle cells of TRPC4 knockout mice was only $34.4{\pm}10.4pA$ (n = 8) at -60 mV. However, slow waves were still present in the knockout mice. Our data suggest that TRPC4 is an essential component of the NSCC activated by muscarinic stimulation in the murine stomach.

Keywords

Acknowledgement

Supported by : Ministry of Information and Communication, SNUH

References

  1. Dickens, E. J., Hirst, G. D. S., and Tomita, T. (1999) Identification of rhythmically active cells in guinea-pig stomach. J. Physiol. 514, 515-531 https://doi.org/10.1111/j.1469-7793.1999.515ae.x
  2. Freichel, M., Suh, S. H., Pfeifer, A., Schweig, U., Trost, C., et al. (2001) Lack of an endothelial store-operated $Ca^{2+}$ current impairs agonist-dependent vasorelaxation in TRP4-/- mice. Nat. Cell Biol. 3, 121-127 https://doi.org/10.1038/35055019
  3. Hirst, G. D. S., Bramich, N. J., Teramoto, N., Suzuki, H., and Edwards, F. R. (2002) Regenerative component of slow waves in the guinea-pig gastric antrum involves a delayed increase in $[Ca^{2+}]_i$ and $Cl^{-}$ channels. J. Physiol. 540, 907-919 https://doi.org/10.1113/jphysiol.2001.014803
  4. Huizinga, J. D., Thuneberg, L., Kluppel, M., Malysz, J., Mikkelsen, H. B., et al. (1995) W/kit gene required for interstitial cells of Cajal and for intestinal pacemaker activity. Nature 373, 347-349 https://doi.org/10.1038/373347a0
  5. Inoue, R., Okada, T., Onoue, H., Hara, Y., Shimizu, S., et al. (2001) The transient receptor potential protein homologue TRP6 is the essential component of vascular ${\alpha}1$-adrenoceptor-activated $Ca^{2+}$- permeable cation channel. Circ. Res. 88, 325-332 https://doi.org/10.1161/01.RES.88.3.325
  6. Ishigawa, S., Komori, K., Nagao, T., and Suzuki, H. (1985) Effects of diltiazem on electrical responses evoked spontaneously or by electrical stimulation in the smooth muscle cells of the guinea-pig stomach. Br. J. Pharmacol. 86, 789-797 https://doi.org/10.1111/j.1476-5381.1985.tb11100.x
  7. Jung, S., Strotmann, R., Schultz, G., and Plant, T. D. (2002) TRPC6 is a candidate channel involved in receptor-stimulated cation currents in A7r5 smooth muscle cells. Am. J. Physiol. Cell Physiol. 82, C347-359
  8. Koh, S. D., Jun, J. Y., Kim, T. W., and Sanders, K. M. (2002) A $Ca^{2+}$-inhibited non-selective cation conductance contributes to pacemaker currents in mouse interstitial cell of Cajal. J. Physiol. 540, 803-814 https://doi.org/10.1113/jphysiol.2001.014639
  9. Komuro, T. and Zhou, D. S. (1996) Anti-c-kit protein immunoreactive cells corresponding to the interstitial cells of Cajal in the guinea-pig small intestine. J. Auton. Nerv. Syst. 61, 169-174 https://doi.org/10.1016/S0165-1838(96)00078-1
  10. Lee, Y. M., Kim, B. J., Kim, H. J., Yang, D. K., Zhu, M. H., et al. (2003) TRPC5 as a candidate for the nonselective cation channel activated by muscarinic stimulation in murine stomach. Am. J. Physiol. Gastrointest. Liver Physiol. 284, G604-G616
  11. Okada, T., Shimizu, S., Wakamori, M., Maeda, A., Kurosaki, T., et al. (1998) Molecular cloning and functional characterization of a novel receptor-activated TRP $Ca^{2+}$ channel from mouse brain. J. Biol. Chem. 273, 10279-10287 https://doi.org/10.1074/jbc.273.17.10279
  12. Philipp, S., Cavalie, A., Freichel, M., Wissenbach, U., Zimmer, S., et al. (1996) A mammalian capacitative calcium entry channel homologous to Drosophila TRP and TRPL. EMBO J. 15, 6166-6171
  13. Philipp, S., Hambrecht, J., Braslavski, L., Schroth, G., Freichel, M., et al. (1998) A novel capacitative calcium entry channel expressed in excitable cells. EMBO J. 17, 4274-4282 https://doi.org/10.1093/emboj/17.15.4274
  14. Philipp, S., Trost, C., Warnat, J., Rautmann, J., Himmerkus, N., et al. (2000) TRP4 (CCE1) protein is part of native calcium release-activated $Ca^{2+}$-like channels in adrenal cells. J. Biol. Chem. 275, 23965-23972 https://doi.org/10.1074/jbc.M003408200
  15. Sanders, K. M. (1992) Ionic mechanisms of electrical rhythmicity in gastrointestinal smooth muscles. Annu. Rev. Physiol. 54, 439-453 https://doi.org/10.1146/annurev.ph.54.030192.002255
  16. Satoh, E., Ono, K., Xu, F., and Iijima, T. (2002) Cloning and functional expression of a novel splice variant of rat TRPC4. Circ. J. 66, 954-958 https://doi.org/10.1253/circj.66.954
  17. Schaefer, M., Plant, T. D., Obukhov, A. G., Hofmann, T., Gudermann, T., et al. (2000) Receptor-mediated regulation of the nonselective cation channels TRPC4 and TRPC5. J. Biol. Chem. 275, 17517-17526 https://doi.org/10.1074/jbc.275.23.17517
  18. Schaefer, M., Plant, T. D., Stresow, N., Albrecht, N., and Schultz, G. (2002) Functional differences between TRPC4 splice variants. J. Biol. Chem. 277, 3752-3759 https://doi.org/10.1074/jbc.M109850200
  19. Suzuki, H., Takano, H., Yamamoto, Y., Komuro, T., Saito, M., et al. (2000) Properties of gastric smooth muscles obtained from mice which lack inositol triphosphate receptor. J. Physiol. 525, 105-111 https://doi.org/10.1111/j.1469-7793.2000.00105.x
  20. Torihashi, S., Fujimoto, T., Trost, C., and Nakayama, S. (2002) Calcium oscillation linked to pacemaking of interstitial cells of Cajal. J. Biol. Chem. 277, 19191-19197 https://doi.org/10.1074/jbc.M201728200
  21. Walker, R. L., Hume, J. R., and Horowitz, B. (2001) Differential expression and alternative splicing of TRP channel genes in smooth muscles. Am. J. Physiol. 280, C1184-C1192
  22. Walker, R. L., Koh, S. D., Sergeant, G. P., Sanders, K. M., and Horowitz, B. (2002) TRPC4 current have properties similar to the pacemaker current in interstitial cells of Cajal. Am. J. Physiol. Cell Physiol. 283, 1637-1645 https://doi.org/10.1152/ajpcell.00266.2002
  23. Ward, S. M., Burns, A. J., Torihashi, S., and Sanders, K. M. (1994) Mutation of the proto-oncogene c-kit blocks development of interstitial cells and electrical rhythmicity in murine intestine. J. Physiol. 480, 91-97
  24. Zhu, M. H., Lee, Y. M., Jin, N. G., So, I., and Kim, K. W. (2003) The transient receptor potential protein homologue TRPC4/5 as a candidate for the nonselective cationic channel activated by muscarinic stimulation in the murine stomach. Neurophysiology 35, 302-307 https://doi.org/10.1023/B:NEPH.0000008790.75165.d6