The long-term objective of this proposal is to study extensively the molecular biology and the biological activity of lipokinins (LPK), natural activators of phospholipase A2 (PLA2) discovered in our laboratory. PLA2 releases arachidonic acid from the cell membrane and therefore is stimulatory to the arachidonic acid cascade. There has been increasing evidence to suggest that the arachidonic acid cascade and its product, the prostaglandins, play major roles in normal development, and that a disturbance of this pathway at any point may lead to teratogenesis. Study of the roles of LPK during normal and abnormal development is therefore of considerable importance. Recently we have succeeded in purifying a 55 kDa LPK. In the proposed study, other LPK will also be purified, and the amino acid sequences and polyclonal antibodies of each will be obtained. Bovine and human LPK cDNAs will be isolated, and the human cDNA will be used to determine the chromosome localization of the LPK gene or genes and for the isolation and characterization of human LPK genomic clones. Production of LPK in prokaryote expression system will be attempted. LPK gene expression will be determined in the developing mouse embryo by Northern blot analysis of RNA isolated from embryos at different developmental stages. Difference in LPK gene expression between normal and drug-treated embryos will also be studied. Biological effects of purified LPK on arachidonic acid release from thymocytes will be tested. Preliminary studies show that partially purified lipokinin prevent hyperglycemia-induced teratogenesis in cultured mouse embryos, and induces growth/differentiation in pre-somite stage mouse embryos in culture. It will be determined whether purified LPK also has these effects on the mouse embryos. The proposed studies should provide important information about the regulations of PLA2 and LPK in normal and abnormal development.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE007939-05
Application #
3221714
Study Section
Oral Biology and Medicine Subcommittee 1 (OBM)
Project Start
1986-12-01
Project End
1994-12-31
Budget Start
1993-01-01
Budget End
1993-12-31
Support Year
5
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of Illinois at Chicago
Department
Type
Schools of Medicine
DUNS #
121911077
City
Chicago
State
IL
Country
United States
Zip Code
60612
Kay, E D; Goldman, A S; Daniel, J C (1990) Common biochemical pathway of dysmorphogenesis in murine embryos: use of the glucocorticoid pathway by phenytoin. Teratog Carcinog Mutagen 10:31-9
Goldman, A S; Katsumata, M; Goto, M P (1988) John Lattimer lecture. Lipokinins: novel phospholipase A2 activators mediate testosterone effects on embryonic genitalia. J Urol 140:1184-8
Goldman, A S; Herold, R; Piddington, R (1988) Inhibition of embryonic palatal shelf horizontalization and medial edge epithelial breakdown by cortisol: role of H-2 in the mouse. J Craniofac Genet Dev Biol 8:135-45
Kay, E D; Goldman, A S; Daniel, J C (1988) Arachidonic acid reversal of phenytoin-induced neural tube and craniofacial defects in vitro in mice. J Craniofac Genet Dev Biol 8:179-86
Goldman, A S (1987) Arachidonic acid and male genital differentiation. Eur J Pediatr 146 Suppl 2:S63-6
Goldman, A S; Van Dyke, D C; Gupta, C et al. (1987) Elevated glucocorticoid receptor levels in lymphocytes of children with the fetal hydantoin syndrome (FHS). Am J Med Genet 28:607-18