The principal aim of this proposal is to isolate novel neuro- and hormonal peptides from the gut and pancreas using a variety of the detection methods and subsequently characterize the chemical, biological and immunochemical properties of the isolated peptides. Large-scale extractions of the tissue materials will be followed by purification using gel permeation, ion exchange and high performance liquid chromatography (HPLC). Specifically, the following gut peptides will be screened: 1) novel peptides which possess the C-terminal amide structure using a chemical assay method; 2) novel peptide which contract or relax smooth muscles using smooth muscle preparations; 3) endogenous gut factors which alter the release of the hormones from pituitary, hypothalamic and intestinal cell cultures. The primary structure of the isolated peptides will be determined using automatic gas-phase sequencing, amino acid analysis and fast- atom-bombardment (FAB) mass spectrometry. Synthetic replicates of the peptides will be prepared using solid-phase synthesizers and will be utilized for generating antibodies, and for pharmacological studies. Further, peptide analogues of peptide YY and neuropeptide Y that have agonist and antagonist activities will be synthesized using a multiple solid-phase peptide synthesis technique. Antibodies raised against the peptides will be used for cellular localization studies using radioimmunoassay and immunohistochemical methods. The structure of precursors of the peptides will be determined by gene cloning techniques and oligonucleotide probes will be utilized for studies of mRNA levels in tissues and for in situ hybridization studies. This study is likely to find a host of new neuropeptides and hormones which have important biological significance and stimulate various physiological, pharmacological, biochemical, and clinical studies. We anticipate that such studies will shed further light on the regulatory mechanisms of the digestive system and will have implications toward understanding the pathogenesis of some digestive diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK039188-05
Application #
3238942
Study Section
General Medicine A Subcommittee 2 (GMA)
Project Start
1987-09-01
Project End
1994-02-28
Budget Start
1991-09-01
Budget End
1994-02-28
Support Year
5
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Stanford University
Department
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
Tatemoto, K; Mann, M J; Shimizu, M (1992) Synthesis of receptor antagonists of neuropeptide Y. Proc Natl Acad Sci U S A 89:1174-8
Funakoshi, A; Miyasaka, K; Kitani, K et al. (1989) Effect of pancreastatin on pancreatic endocrine function in the conscious rat. Regul Pept 24:225-31
Miyasaka, K; Funakoshi, A; Nakamura, R et al. (1989) Effects of porcine pancreastatin on postprandial pancreatic exocrine secretion and endocrine functions in the conscious rat. Digestion 43:204-11
Nakano, I; Funakoshi, A; Miyasaka, K et al. (1989) Isolation and characterization of bovine pancreastatin. Regul Pept 25:207-13
Lagny-Pourmir, I; Amiranoff, B; Lorinet, A M et al. (1989) Characterization of galanin receptors in the insulin-secreting cell line Rin m 5F: evidence for coupling with a pertussis toxin-sensitive guanosine triphosphate regulatory protein. Endocrinology 124:2635-41
Funakoshi, A; Miyasaka, K; Nakamura, R et al. (1989) Inhibitory effect of pancreastatin on pancreatic exocrine secretion in the conscious rat. Regul Pept 25:157-66
Lorinet, A M; Tatemoto, K; Laburthe, M et al. (1989) Pancreastatin inhibits insulin release from Rin m 5F cells: reversion by pertussis toxin. Eur J Pharmacol 160:405-7
Gunion, M W; Rosenthal, M J; Tatemoto, K et al. (1989) Intracranial microinfusion of pancreastatin elevates blood glucose, free fatty acids, and corticosterone in rats. Brain Res 485:251-7
Flood, J F; Morley, J E; Tatemoto, K (1988) Effects of systemic pancreastatin on memory retention. Peptides 9:1077-80
Tatemoto, K; Nakano, I; Makk, G et al. (1988) Isolation and primary structure of human peptide YY. Biochem Biophys Res Commun 157:713-7