| Co-Investigator(Kenkyū-buntansha) |
YOKOGOSHI Yutaka SCHOOL OF MEDICINE, THE UNIVERSITY OF TOKUSHIMA, INSTRUCTOR, 医学部・内科学第一講座, 助手 (00253203)
BANDO Hiroshi UNIVERSITY OF TOKUSHIMA HOSPITAL, INSTRUCTOR, 医学部・附属病院第一内科, 助手 (60228690)
新谷 保実 徳島大学, 医学部・附属病院, 助手 (10235773)
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| Budget Amount *help |
¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 1993: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1992: ¥4,700,000 (Direct Cost: ¥4,700,000)
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| Research Abstract |
1) Solubilization of somatostatin receptor Somatostatin receptor was solubilized from pig pituitary and human placental membrane fractions using 10 mM CHAPS.The solubilized receptor cross-linked with isotope labelled ligand using 1 mM DSS showed multiple band on SDS-PAGE and was eluted from Sephacryl-S300 column at the void volume, indicating high molecular complex formation.
2) PCR-cloning of somatostatin receptors genomic clones for somatostatin receptor typel (SSTR1) and somatostatin receptor type2 (SSTR2) were obtained by PCR-cloning. Southern blot analysis of SSTR1 gene showed a single fragment of EcoRI, PstI, and BamHI digestioin of 20, 2.7, and 7Kb, respectively. Southern blot analysis of SSTR2 gene showed a single fragment of EcoRI, PstI, and BamHI digestioin of 30, 11, and 30 Kb, respectively.
3) Cloning of porcine somatostatin receptor type 2 gene A clone of pig somatostatin receptor type 2 gene (pSSTR2) was cloned form pig genomic library. The cloned DNA contained an open read
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ing frame of 1107 bp and encoded 369 amino acids containing 7 hydrophobic domains suggesting G-protein coupled receptor with 7 transmembrane domains.
3) Expression of somatostatin receptors High level of expression of SSTR1 was found in stomach ad small intestine. On the other hand, high level of expression of SSTR2 was found in the brain, the pituitary and peripheral lymphocytes. Both receptor showed variable expression in endocrine tumors. In our series, SSTR2 mRNA expression was detected in the thyroid tissues of the patients with Basedow's disease, thyroid adenoma or thyroid papillary carcinoma. SSTR1 mRNA expression was detected in tumors of thyroid medullary carcinoma and pheochromocytoma.
4) Point mutation of somatostatin receptors SSCP and TGGE analysis detected various mutated band in tissues of thyroid adenoma, adenomatous goiter, thyroid papillary carcinoma and ACTH producing cell line. A missense mutation of TGG to TGA at codon 188 of SSTR2 gene was found in an ACTH-producing cell line (COR-L103). Less
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