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A minor role for Ca2+ sensitization in sustained contraction through activation of muscarinic receptor in circular muscle of rat distal colon

  • Gastrointestinal Function
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Abstract

We previously demonstrated that Ca2+ sensitization has an essential role for carbachol-induced contraction in the longitudinal muscle of the rat distal colon. In the present study, we extended these studies to clarify the role of Ca2+ sensitization in contraction induced by the activation of muscarinic receptors in the circular muscle of the rat distal colon. Carbachol induced a rapid phasic contraction followed by a sustained contraction that was significantly lower than the phasic and was superimposed with the rhythmic contractions. The extent of increase in intracellular Ca2+ concentration that was measured simultaneously with tension recording was dissociated from the phasic contraction, whereas it exhibited to a similar extent as sustained contraction. In α-toxin-permeabilized preparations, Ca2+ induced contraction comprising a rapid phasic and a subsequent low sustained component. After Ca2+-induced sustained contraction reached a constant level, guanosine triphosphate (GTP) addition resulted in the enhancement of contractile force in a concentration-dependent manner. Carbachol in the presence of GTP caused a further minimal increase in tension (Ca2+ sensitization). Chelerythrine, a protein kinase C (PKC) inhibitor, inhibited carbachol-induced Ca2+ sensitization but not GTP-induced Ca2+ sensitization. In contrast, Y-27632, a Rho kinase inhibitor, inhibited GTP-induced Ca2+ sensitization but not that induced by carbachol. Phorbol 12,13-dibutyrate, a PKC activator, increased the sustained contraction. These results suggest that the activation of muscarinic receptor with carbachol induces Ca2+ sensitization via activation of PKC, but this action is minor in the circular muscle of the rat distal colon as a result of limited coupling between muscarinic receptors and Ca2+ sensitization via the PKC pathway.

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Abbreviations

PDBu:

phorbol 12,13-dibutyrate

PKC:

protein kinase C

[Ca2+]i:

intracellular Ca2+ concentration

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Acknowledgments

We thank Dr. G.S. Drummond (InfaCare Pharmaceutical Corporation, PA, USA) for critical reading of this manuscript. This work was supported in part by Grants-in-Aid for Scientific Research (C) from the Japan Society for the Promotion of Science, and by scholarships from Ono Pharmaceutical.

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  1. Laboratory of Veterinary Pharmacology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai Osaka, 599-8531, Japan

    Tadayoshi Takeuchi, Hidemitsu Nakajima, Fumiaki Hata & Yasu-Taka Azuma

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  1. Tadayoshi Takeuchi
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Correspondence to Tadayoshi Takeuchi.

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Takeuchi, T., Nakajima, H., Hata, F. et al. A minor role for Ca2+ sensitization in sustained contraction through activation of muscarinic receptor in circular muscle of rat distal colon. Pflugers Arch - Eur J Physiol 454, 565–574 (2007). https://doi.org/10.1007/s00424-007-0221-7

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