The objective of this proposal is to investigate the functional role of the lipid mediators generated by the 12/15-lipoxygenase (12/15-LOX) enzymes in the control of uterine function during implantation. The 12/15-LOX catalyzes the stereo-specific oxygenation of the 20-carbon polyunsaturated fatty acids, arachidonic acid and linoleic acid, into a complex series of derivatives, HETEs and HODES. Recent studies, using high-density oligonucleotide microarrays, revealed that progesterone markedly induces the synthesis of mRNAs encoding the 12/15-LOX family members, leukocyte-12/15-LOX and epidermal-12/15-LOX, in the surface epithelium of pregnant uterus precisely at the time of implantation. Administration of a LOX-specific inhibitor markedly inhibited steroid hormone-regulated vascular permeability during delayed implantation in mice, indicating an important role of these enzymes in creating a receptive uterus. The major goals of this proposal are to: 1. Identify the gene networks that mediate the physiological effects of these lipid metabolites in the uterus during implantation. The 12/15-LOX-regulated gene pathways will be identified by DNA microarray. The spatio-temporal expression of these genes in the peri-implantation uterus will be analyzed and their function will be determined by a newly developed antisense oligonucleotide strategy. 2. Investigate the role of PPARgamma in the 12/15-LOX signaling pathway during implantation. The 12/15-LOX-derived metabolites of arachidonic or linoleic acid activate gene transcription by the peroxisome proliferator-activated receptor gamma (PPARgamma) in cell-based assays. PPARgamma expression is also induced in the uterus in the peri-implantation period. We will test whether PPARgamma is indeed the endogenous receptor for the 12/15-LOX-derived metabolites in the peri-implantation uterus and identify its target genes in this tissue. The role of PPARgamma will be further investigated by generating a conditional knockout of its gene in the uterus and by analyzing the functional consequences of the loss-of-function mutation of this receptor during implantation. Collectively, the proposed experiments will test the hypothesis that a novel signaling pathway, involving 12/15-LOX-derived lipid mediators, PPARgamma, and its downstream target genes, regulates critical events during implantation.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD044611-04
Application #
7055336
Study Section
Reproductive Biology Study Section (REB)
Program Officer
Yoshinaga, Koji
Project Start
2003-07-01
Project End
2008-04-30
Budget Start
2006-05-01
Budget End
2008-04-30
Support Year
4
Fiscal Year
2006
Total Cost
$299,295
Indirect Cost
Name
University of Illinois Urbana-Champaign
Department
Physiology
Type
Schools of Arts and Sciences
DUNS #
041544081
City
Champaign
State
IL
Country
United States
Zip Code
61820
Kim, Jaeyeon; Bagchi, Indrani C; Bagchi, Milan K (2009) Signaling by hypoxia-inducible factors is critical for ovulation in mice. Endocrinology 150:3392-400
Laws, Mary J; Taylor, Robert N; Sidell, Neil et al. (2008) Gap junction communication between uterine stromal cells plays a critical role in pregnancy-associated neovascularization and embryo survival. Development 135:2659-68
Kim, Jaeyeon; Sato, Marcey; Li, Quanxi et al. (2008) Peroxisome proliferator-activated receptor gamma is a target of progesterone regulation in the preovulatory follicles and controls ovulation in mice. Mol Cell Biol 28:1770-82
Li, Quanxi; Kannan, Athilakshmi; Wang, Wei et al. (2007) Bone morphogenetic protein 2 functions via a conserved signaling pathway involving Wnt4 to regulate uterine decidualization in the mouse and the human. J Biol Chem 282:31725-32
Palanisamy, Gopinath S; Cheon, Yong-Pil; Kim, Jaeyeon et al. (2006) A novel pathway involving progesterone receptor, endothelin-2, and endothelin receptor B controls ovulation in mice. Mol Endocrinol 20:2784-95
Bagchi, Milan K; Mantena, Srinivasa R; Kannan, Athilakshmi et al. (2006) Control of uterine cell proliferation and differentiation by C/EBPbeta: functional implications for establishment of early pregnancy. Cell Cycle 5:922-5
Mantena, Srinivasa Raju; Kannan, Athilakshmi; Cheon, Yong-Pil et al. (2006) C/EBPbeta is a critical mediator of steroid hormone-regulated cell proliferation and differentiation in the uterine epithelium and stroma. Proc Natl Acad Sci U S A 103:1870-5
Li, Quanxi; Bagchi, Milan K; Bagchi, Indrani C (2006) Identification of a signaling pathway involving progesterone receptor, calcitonin, and tissue tranglutaminase in Ishikawa endometrial cells. Endocrinology 147:2147-54
Bagchi, Indrani C; Li, Quanxi; Cheon, Yong-Pil et al. (2005) Use of the progesterone receptor antagonist RU 486 to identify novel progesterone receptor-regulated pathways in implantation. Semin Reprod Med 23:38-45
Cheon, Yong-Pil; DeMayo, Francesco J; Bagchi, Milan K et al. (2004) Induction of cytotoxic T-lymphocyte antigen-2beta, a cysteine protease inhibitor in decidua: a potential regulator of embryo implantation. J Biol Chem 279:10357-63

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