Intracellular calcium mobilization of the aganglionic intestine in the endothelin B receptor gene -deficient rat

Takanori Nakatsuji, Satoshi Ieiri, Kouji Masumoto, Junko Akiyoshi, Tomoaki Taguchi, Sachiyo Suita

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background and Aim: Up to now, numerous reports have analyzed the pathogenesis of Hirschsprung's disease (HD) by means of physiologic, pathologic, or molecular biologic methods. However, very little is still known about the smooth muscle cell itself. The endothelin B receptor gene-deficient (EDNRB(-/-)) rat, which is suitable for research of HD, has an aganglionic segment of the total colon. Our purpose is to investigate the myogenic mechanisms using simultaneous measurements of the intracellular Ca2+ concentration ([Ca2+]i) and tension and reverse transcriptase polymerase chain reaction for L-type Ca2+ channel (L-VOC) expression. Methods: The muscle strips of the rat distal colon were loaded with a Ca2+ indicator dye, fura-PE3/AM, for 3 to 4 hours. The changes in the fluorescence intensity of Ca2+-fura-PE3 complex of the strips were monitored with a front surface fluorometer (CAM-230). The fluorescence intensities at 340- and 380-nm excitation and their ratio (F340/F380) were recorded as the level of [Ca2+]i. The comparison of L-VOC α1c subunit messenger RNA (mRNA) expression in both wild and homozygous rat was performed by reverse transcriptase polymerase chain reaction. Results: The peak levels of force development induced by carbachol were 139.1% ± 5.0% in EDNRB(-/-) rat, whereas the peak levels were 242.1% ± 27.7% in EDNRB(+/+) rat. The changes in the [Ca2+]i elevation induced by carbachol were 101.7% ± 12.2% in the homozygous rat, whereas these were 143.8% ± 8.9% in the wild-type rat. Both results in the homozygous rat significantly decreased in comparison with those of the wild rat (P < .05). The expression of the L-VOC channel mRNA also decreased in the homozygous rat. Conclusions: This is the first report to show the [Ca2+]i mobilization in the smooth muscles of the rat model of HD. The decrease in both [Ca2+]i and force development was thus considered to be due to the decrease in the Ca2+ channel expression.

Original languageEnglish
Pages (from-to)1663-1670
Number of pages8
JournalJournal of Pediatric Surgery
Volume42
Issue number10
DOIs
Publication statusPublished - Oct 1 2007

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Endothelin B Receptors
Intestines
Calcium
Genes
Hirschsprung Disease
Carbachol
Reverse Transcriptase Polymerase Chain Reaction
Colon
Fluorescence
Messenger RNA
Smooth Muscle Myocytes
Smooth Muscle
Coloring Agents

All Science Journal Classification (ASJC) codes

  • Surgery

Cite this

Intracellular calcium mobilization of the aganglionic intestine in the endothelin B receptor gene -deficient rat. / Nakatsuji, Takanori; Ieiri, Satoshi; Masumoto, Kouji; Akiyoshi, Junko; Taguchi, Tomoaki; Suita, Sachiyo.

In: Journal of Pediatric Surgery, Vol. 42, No. 10, 01.10.2007, p. 1663-1670.

Research output: Contribution to journalArticle

Nakatsuji, Takanori ; Ieiri, Satoshi ; Masumoto, Kouji ; Akiyoshi, Junko ; Taguchi, Tomoaki ; Suita, Sachiyo. / Intracellular calcium mobilization of the aganglionic intestine in the endothelin B receptor gene -deficient rat. In: Journal of Pediatric Surgery. 2007 ; Vol. 42, No. 10. pp. 1663-1670.
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abstract = "Background and Aim: Up to now, numerous reports have analyzed the pathogenesis of Hirschsprung's disease (HD) by means of physiologic, pathologic, or molecular biologic methods. However, very little is still known about the smooth muscle cell itself. The endothelin B receptor gene-deficient (EDNRB(-/-)) rat, which is suitable for research of HD, has an aganglionic segment of the total colon. Our purpose is to investigate the myogenic mechanisms using simultaneous measurements of the intracellular Ca2+ concentration ([Ca2+]i) and tension and reverse transcriptase polymerase chain reaction for L-type Ca2+ channel (L-VOC) expression. Methods: The muscle strips of the rat distal colon were loaded with a Ca2+ indicator dye, fura-PE3/AM, for 3 to 4 hours. The changes in the fluorescence intensity of Ca2+-fura-PE3 complex of the strips were monitored with a front surface fluorometer (CAM-230). The fluorescence intensities at 340- and 380-nm excitation and their ratio (F340/F380) were recorded as the level of [Ca2+]i. The comparison of L-VOC α1c subunit messenger RNA (mRNA) expression in both wild and homozygous rat was performed by reverse transcriptase polymerase chain reaction. Results: The peak levels of force development induced by carbachol were 139.1{\%} ± 5.0{\%} in EDNRB(-/-) rat, whereas the peak levels were 242.1{\%} ± 27.7{\%} in EDNRB(+/+) rat. The changes in the [Ca2+]i elevation induced by carbachol were 101.7{\%} ± 12.2{\%} in the homozygous rat, whereas these were 143.8{\%} ± 8.9{\%} in the wild-type rat. Both results in the homozygous rat significantly decreased in comparison with those of the wild rat (P < .05). The expression of the L-VOC channel mRNA also decreased in the homozygous rat. Conclusions: This is the first report to show the [Ca2+]i mobilization in the smooth muscles of the rat model of HD. The decrease in both [Ca2+]i and force development was thus considered to be due to the decrease in the Ca2+ channel expression.",
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AU - Nakatsuji, Takanori

AU - Ieiri, Satoshi

AU - Masumoto, Kouji

AU - Akiyoshi, Junko

AU - Taguchi, Tomoaki

AU - Suita, Sachiyo

PY - 2007/10/1

Y1 - 2007/10/1

N2 - Background and Aim: Up to now, numerous reports have analyzed the pathogenesis of Hirschsprung's disease (HD) by means of physiologic, pathologic, or molecular biologic methods. However, very little is still known about the smooth muscle cell itself. The endothelin B receptor gene-deficient (EDNRB(-/-)) rat, which is suitable for research of HD, has an aganglionic segment of the total colon. Our purpose is to investigate the myogenic mechanisms using simultaneous measurements of the intracellular Ca2+ concentration ([Ca2+]i) and tension and reverse transcriptase polymerase chain reaction for L-type Ca2+ channel (L-VOC) expression. Methods: The muscle strips of the rat distal colon were loaded with a Ca2+ indicator dye, fura-PE3/AM, for 3 to 4 hours. The changes in the fluorescence intensity of Ca2+-fura-PE3 complex of the strips were monitored with a front surface fluorometer (CAM-230). The fluorescence intensities at 340- and 380-nm excitation and their ratio (F340/F380) were recorded as the level of [Ca2+]i. The comparison of L-VOC α1c subunit messenger RNA (mRNA) expression in both wild and homozygous rat was performed by reverse transcriptase polymerase chain reaction. Results: The peak levels of force development induced by carbachol were 139.1% ± 5.0% in EDNRB(-/-) rat, whereas the peak levels were 242.1% ± 27.7% in EDNRB(+/+) rat. The changes in the [Ca2+]i elevation induced by carbachol were 101.7% ± 12.2% in the homozygous rat, whereas these were 143.8% ± 8.9% in the wild-type rat. Both results in the homozygous rat significantly decreased in comparison with those of the wild rat (P < .05). The expression of the L-VOC channel mRNA also decreased in the homozygous rat. Conclusions: This is the first report to show the [Ca2+]i mobilization in the smooth muscles of the rat model of HD. The decrease in both [Ca2+]i and force development was thus considered to be due to the decrease in the Ca2+ channel expression.

AB - Background and Aim: Up to now, numerous reports have analyzed the pathogenesis of Hirschsprung's disease (HD) by means of physiologic, pathologic, or molecular biologic methods. However, very little is still known about the smooth muscle cell itself. The endothelin B receptor gene-deficient (EDNRB(-/-)) rat, which is suitable for research of HD, has an aganglionic segment of the total colon. Our purpose is to investigate the myogenic mechanisms using simultaneous measurements of the intracellular Ca2+ concentration ([Ca2+]i) and tension and reverse transcriptase polymerase chain reaction for L-type Ca2+ channel (L-VOC) expression. Methods: The muscle strips of the rat distal colon were loaded with a Ca2+ indicator dye, fura-PE3/AM, for 3 to 4 hours. The changes in the fluorescence intensity of Ca2+-fura-PE3 complex of the strips were monitored with a front surface fluorometer (CAM-230). The fluorescence intensities at 340- and 380-nm excitation and their ratio (F340/F380) were recorded as the level of [Ca2+]i. The comparison of L-VOC α1c subunit messenger RNA (mRNA) expression in both wild and homozygous rat was performed by reverse transcriptase polymerase chain reaction. Results: The peak levels of force development induced by carbachol were 139.1% ± 5.0% in EDNRB(-/-) rat, whereas the peak levels were 242.1% ± 27.7% in EDNRB(+/+) rat. The changes in the [Ca2+]i elevation induced by carbachol were 101.7% ± 12.2% in the homozygous rat, whereas these were 143.8% ± 8.9% in the wild-type rat. Both results in the homozygous rat significantly decreased in comparison with those of the wild rat (P < .05). The expression of the L-VOC channel mRNA also decreased in the homozygous rat. Conclusions: This is the first report to show the [Ca2+]i mobilization in the smooth muscles of the rat model of HD. The decrease in both [Ca2+]i and force development was thus considered to be due to the decrease in the Ca2+ channel expression.

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