Involvement of Thromboxane A2 in the Modulation of Pacemaker Activity of Interstitial Cells of Cajal of Mouse Intestine

Jin Ho Kim; Soo Jin Choi; Cheol Ho Yeum; Pyung Jin Yoon; Seok Choi; Jae Yeoul Jun

doi:10.4196/kjpp.2008.12.1.25

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Kim, Choi, Yeum, Yoon, Choi, and Jun: Involvement of Thromboxane A2 in the Modulation of Pacemaker Activity of Interstitial Cells of Cajal of Mouse Intestine

Original Article

Korean J Physiol Pharmacol 2008;12(1):25-30.

Published online: 17 February 2008

DOI: https://doi.org/10.4196/kjpp.2008.12.1.25

Involvement of Thromboxane A₂ in the Modulation of Pacemaker Activity of Interstitial Cells of Cajal of Mouse Intestine

Jin Ho Kim¹, Soo Jin Choi², Cheol Ho Yeum³, Pyung Jin Yoon³, Seok Choi³, Jae Yeoul Jun³

¹Department of Neurology, College of Medicine, Chosun University, Gwangju 501-759, Korea

²Department of Radiology, Gacheon University Gil Medical Center, Guwol-dong, Incheon 405-760, Korea

³Department of Physiology, College of Medicine, Chosun University, Gwangju 501-759, Korea

Corresponding Author: Jae Yeoul Jun, Department of Physiology, College of Medicine, Chosun University, 375, Seoseok-dong, Dong-gu, Gwangju 501-759, Korea. (Tel) 82-62-230-6412, (Fax) 82-62-232-4943, (E-mail) jyjun@ chosun.ac.kr

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Although many studies show that thromboxane A₂ (TXA₂) has the action of gastrointestinal (GI) motility using GI muscle cells and tissue, there are no reports on the effects of TXA₂ on interstitial cells of Cajal (ICC) that function as pacemaker cells in GI tract. So, we studied the modulation of pacemaker activities by TXA₂ in ICC with whole cell patch-clamp technique. Externally applied TXA₂ (5μM) produced membrane depolarization in current-clamp mode and increased tonic inward pacemaker currents in voltage-clamp mode. The tonic inward currents by TXA₂ were inhibited by intracellular application of GDP-β-S. The pretreatment of ICC with Ca²⁺ free solution and thapsigargin, a Ca²⁺-ATPase inhibitor in endoplasmic reticulum, abolished the generation of pacemaker currents and suppressed the TXA₂-induced tonic inward currents. However, chelerythrine or calphostin C, protein kinase C inhibitors, did not block the TXA₂-induced effects on pacemaker currents. These results suggest that TXA₂ can regulate intestinal motility through the modulation of ICC pacemaker activities. This modulation of pacemaker activities by TXA₂ may occur by the activation of G protein and PKC independent pathway via extra and intracellular Ca²⁺ modulation.

Keywords: Thromboxane A₂ (TXA₂), Interstitial cells of Cajal (ICC), Pacemaker currents, Intestinal motility

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Fig. 1.

The effects of TXA2 on pacemaker potentials and pacemaker currents recorded in cultured ICC from mouse small intestine (A) Pacemaker potentials of ICC which were exposed to TXA2 (5 μM) in the current-clamping mode (I=0). Vertical solid line scales amplitude of pacemaker potential and horizontal solid line scales for duration of recording (s) pacemaker potentials. (B), (C), and (D) Pacemaker currents of ICC recorded at a holding potential of −70 mV, when exposed to various concentrations of TXA2 (1, 3, and 5 μM). The dotted lines indicate zero current levels. Vertical solid line scales amplitude of pacemaker current and horizontal solid line scales duration of recording (s) pacemaker currents. The responses to TXA2 are summarized in (E), (F) and (G). The bars represent mean values±SE. ∗∗Significantly different from the untreated control (Con) (p < 0.01).

Fig. 2.

The effects of GDP-β-S in response to TXA₂ induced pacemaker currents from ICC of mouse small intestine (A) Pacemaker currents from ICC exposed to TXA₂ (5 μM) in the presence of GDP-β-S (1 mM) in the pipette. The tonic inward currents with suppressed amplitudes and frequency induced by TXA₂ were blocked by internally applied GDP-β-S (1 mM). The dotted lines indicate the zero current levels. The effects of TXA2 in the presence of GDP-β-S are summarized in (B). Vertical solid line scales amplitude of pacemaker current and horizontal solid line scales for duration of recording (s) pacemaker currents. Bars represent mean values ± SE. The effects of GDP-β-S on TXA₂-induced pacemaker currents were significantly different from the TXA₂-induced pacemaker currents (p < 0.01). The bars represent mean values±SE. ∗∗Significantly different from the untreated control (Con) (p < 0.01).

Fig. 3.

The effects of an external Ca²⁺-free solution or thapsigargin on the TXA₂-induced response on pacemaker currents in cultured ICC from mouse small intestine (A) External Ca²⁺-free solution abolished the generation of pacemaker currents. Under this condition, the TXA₂ (5μM)-induced tonic inward currents were blocked. (C) Thapsigargin (5μM) abolished the generation of pacemaker currents. Thapsigargin also blocked the TXA₂ (5μM)-induced tonic inward currents. The dotted lines indicate the zero current levels. Responses to the TXA₂ in the external Ca²⁺-free solution and in the presence of thapsigargin are summarized in (B) and (D). Vertical solid line scales amplitude of pacemaker current and horizontal solid line scales duration of recording (s) pacemaker current. The bars represent mean values ± SE. ∗∗Significantly different from the untreated Control (Con) (p < 0.01).

Fig. 4.

The effects of chelerythrine or calphostin C on the TXA₂-induced response on pacemaker currents in cultured ICC from mouse small intestine (A), (B) Pacemaker currents of ICC exposed to TXA₂ (10 μM) in the presence of chelerythrine (1μM) or calphostin C (0.1μM). In this condition, the TXA₂ caused tonic inward currents. The dotted lines indicate the zero current levels. Responses to the TXA₂ in the presence of chelerythrine or calphostin C are summarized in (C). Vertical solid line scales amplitude of pacemaker current and horizontal solid line scales duration of recording (s) pacemaker current. The bars represent mean values ± SE.

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