| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1989: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1988: ¥4,700,000 (Direct Cost: ¥4,700,000)
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| Research Abstract |
Synthesis of brain-gut hormones: We have synthesized several brain- gut hormones and the related peptides containing 15-20 amino acid residues. Syntheses of these peptides were performed according to solid phase technology using an automated peptide synthesized, Beckman Model 990. A protected peptide chain was constructed on a PAM resin (Applied Biosystems). The purity of the synthetic peptides was proved by amino acid analyses of both acid hydrolysates and AP-M digests of the peptides, analytical C_<18> reverse-phase HPLC and the sequence analysis of the peptide using an automated gas-phase sequencer (Applied Biosystem 470A)., We have synthesized in the present study several new brain-gut hormones such as porcine and rat galanin, helodermin, guinea pig VIP, rat PHI, human glucagon-like peptide-1 (GLP-1), human NPY human and rat proLH-RH-related peptide such as gonadotropin releasing hormone-associated peptide (GAP).
Production of antisera against brain-gut hormoned: GLP-1 for productio
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n of GLP-1 specific antisera, synthetic GLP-1(7-22) in porcine thyroglobulin-conjugated form and GLP-1(1-35) in non-conjugated form were selected as immunogens and rabbits were immunized with each of the immunogens. The resulting antisera, anti-GLP-1(7-22) serum RO322 and anti-GLP-1(1-35) serum RO351, were used for the development of radioimmunoassay (RIA)-systems region-specific for GLP-1.
Galanin: Since galanin has structural similarities in its C-terminal region to tachykinins, a monoclonal antibody against synthetic porcine galanin(1-15) was successfully produced especially for immunohistochemical study. In fact, this monoclonal antibody was found to recognize specifically galanin-producing cells and did not cross-react with any of tachykinins.
Pro-LHRH related peptide: LHRH precursor has been known to contain LHRH and a GAP with 56 amino acid peptide. Using synthetic GAP(34-56) and proLHRH(1-26), we have developed human pro-LHRH carboxy- and amino- terminal specific RIA with the production of anti GAP(34-56) serum(RO242) and anti-proLHRH(1-26) serum (RO620). An antiserum against synthetic rat GAP(28-51) (RO246) was also generated in a rabbits. Less
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