| Project/Area Number |
09671841
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
小児外科
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| Research Institution | Kansai Medical University |
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Principal Investigator |
HAMADA Yoshinori Kansai Medical University Faculty of Medicine Associate Professor, 医学部, 助教授 (00172982)
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| Co-Investigator(Kenkyū-buntansha) |
SATO Masahito Kansai Medical University Faculty of Medicine Assistant, 医学部, 助手 (30278626)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1999: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Nitric oxide / Endothelin / Hirschsprung's disease / EDNRB gene |
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| Research Abstract |
Immunohistochemistry of bNOS, eNOS, and iNOS was performed on resected specimen of Hirschsprung's disease. Localization of eNOS and iNOS was observed in vascular endothelium and in submucosal layer to mucosa, respectively in the control specimen, which were obtained from colostomy in patients with anorectal malformation. In the Hirschsprung's disease, no significant differences were observed between aganglionic and normoganglionic segment, and findings of localization were as same as those in control bowel. NADPH immunohistochemistry also showed nearly same results as reported before. However, bNOS localization was found in the nerve bundles appeared in the intermuscular layer of the aganglionic segment.
Immunohistochemistry endothelin 3 and endothelin 1 receptor was also performed. In the control bowel, endothelin 3 and endothelin 1 receptor were strongly localized in the submucosal plexus and mucosal muscle layer and also in the both muscle layers and intermuscular plexus. Affected bowel of Hirschsprung's disease did not show any differences in immunohistochemical findings as compared with those in controls.
Autoradiography of endothelin A receptor (EDNRA) and endothelin B receptor (EDNRB) was performed using freshly frozen section of resected bowels. In the control bowel, EDNRA was localized weakly in the submucosal layer and EDNRB was found not only in the proper muscle layer but also in both the intermuscular and submucosal plexus. In the affected segment of Hirschsprung's disease, EDNRB was localized in the nerve bundles in the intermuscular layer.
Mutational analysis of EDNRB gene was performed by PCR-SSCP method using peripheral leucocytes from Hirschsprung's disease. No mutation was found in all seven exons of the EDNRB gene even in a patient with congenital central hypoventilation syndrome.
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