Evidences in the literature indicate the existence of an embryonic insulin which could play a role in controlling fetal growth at stages when pancreas has not yet become functional. We have developed a mesodermal cell line called 1246-3A derived from C3H mouse teratoma which produces and secretes in culture a growth factor called insulin-related factor (IRF) which is immunologically and biochemically similar to pancreatic insulin. IRF can be purified to homogeneity from 1246-3A conditioned medium in a very rapid, efficient procedure which can be easily scaled up. It includes CG50 chromatography, immunoaffinity chromatography and reverse phase HPLC. As pure IRF can be obtained in Mug quantity it will be possible to determine its structure and study the regulation of its secretion and its expression in vitro and in vivo. These experiments are performed in order to investigate the possible involvement of IRF as a growth factor during the pre-pancreatic stage of embryonic development. More specifically, IRF amino acid sequence will be determined by gas phase sequencing. Based on this data, oligonucleotide sequences corresponding to IRF sequence will be synthesized and used to clone cDNA encoding for IRF from a 1246-3A cDNA library, in order to investigate the regulation of the gene coding for IRF during embryonic development. Partial cDNA sequence analysis will be performed in order to compare it to the pancreatic insulin gene. This system presented in this proposal should prove a suitable one to the long-term goal of investigating the nature and function of emryonic extra-pancreatic insulin produced by mesodermal tissue and to understand its function in vivo as a growth factor during development.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM037606-03
Application #
3293013
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1986-12-01
Project End
1989-11-30
Budget Start
1988-12-01
Budget End
1989-11-30
Support Year
3
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Adirondack Biomedical Research Institute
Department
Type
DUNS #
City
Lake Placid
State
NY
Country
United States
Zip Code
12946
Litthauer, D; Serrero, G (1992) The primary culture of mouse adipocyte precursor cells in defined medium. Comp Biochem Physiol A Comp Physiol 101:59-64
Yamada, Y; Serrero, G (1989) Characterization of transforming growth factors produced by the insulin-independent teratoma-derived cell line 1246-3A. J Cell Physiol 140:254-63
Gazzano, H; Serrero, G (1988) Identification of insulin receptors on the insulin-independent variant 1246-3A cell line. J Cell Physiol 136:348-54