Crosstalk between the immune and reproductive systems dramatically impacts fertility. In particular, studies have shown that a tight spatiotemporal control of leukocyte infiltration into the ovary is pivotal for successful ovulation. Once infiltrated, the leukocytes differentiate into a cohort of pro- or anti-infiammatory cells in the preovulatory ovary and play critical roles in ovulation and luteal formation. In this study, we will test the hypothesis that progesterone (P4) and prostaglandins (PG) play pivotal roles in recruiting leukocytes into the preovulatory ovary.
Aim 1. To determine the mechanism(s) by which P4 and PG regulate WBC infiltration. We will first determine the roles of P4 and PG on WBC infiltration by treating cycling adult rats with RU486, indomethacin and vehicle, and measuring the effect on the numbers, populations and localization of infiltrating WBC by flow cytometry and immunohistochemistry together with a functional analysis of the expression of chemokines, cytokines, their receptors and cell adhesion molecules (CAMs) in serum, on ovarian cells and WBC by ELISA and qPCR.
Aim 2. To determine whether P4 and PG regulate WBC infiltration in primate ovary. In this Aim, we will investigate P4 and PG regulated WBC infiltration in primates. Similar to rodents, primates, including humans also require P4 and PG for normal ovulation. Therefore, we hypothesize that P4 and PG will regulate WBC infiltration in the primate ovary. The actions of P4 and PG will be inhibited by treating primates (Cynomolgous monkeys) with trilostane (3|3HSD inhibitor;inhibits progesterone synthesis), celecoxib (selective COX-2 inhibitor) or vehicle under ovarian stimulation. The effects on the numbers, populations and localization of WBC together with the expression of infiammatory mediators will be determined. Upon the identification of the P4- and PG-regulated WBC subtypes and infiammatory mediators in the monkey, the localization and expression patterns of their homologs will be determined in the human ovary.
Aim 3. To determine the regulatory mechanism of preovulatory splenic WBC release. We will identify the trigger(s) and the mechanism of action of splenic WBC release from spleen. We hypothesize that either LH, decreased concentration of circulating WBC or angiotensin-ll (the only known trigger of splenic leukocyte release) act as the trigger. To identify the trigger(s), ovariectomized rats will be treated with candidate triggers (LH and angiotensin-ll) or artificially reduce the concentration of circulating WBC by perfusing rats with plasma. The effects on splenic WBC release will be measured by counting WBC numbers in spleen and peripheral blood.

Public Health Relevance

Chronic anovulation affects up to 15% of the female US population, many of the causes not being accounted for. This study will identify a novel cause of anovulation resulting from defective ovarian inflammation, which will lead to develop a better treatment option for an ovulatory disorder.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Program Projects (P01)
Project #
1P01HD071875-01A1
Application #
8609429
Study Section
Special Emphasis Panel (ZHD1-DRG-D (41))
Project Start
Project End
Budget Start
2014-09-19
Budget End
2015-06-30
Support Year
1
Fiscal Year
2014
Total Cost
$136,534
Indirect Cost
$12,500
Name
University of Kentucky
Department
Type
DUNS #
939017877
City
Lexington
State
KY
Country
United States
Zip Code
40506
Cacioppo, Joseph A; Lin, Po-Ching Patrick; Hannon, Patrick R et al. (2017) Granulosa cell endothelin-2 expression is fundamental for ovulatory follicle rupture. Sci Rep 7:817
Puttabyatappa, Muraly; Al-Alem, Linah F; Zakerkish, Farnosh et al. (2017) Induction of Tissue Factor Pathway Inhibitor 2 by hCG Regulates Periovulatory Gene Expression and Plasmin Activity. Endocrinology 158:109-120
Choi, Yohan; Wilson, Kalin; Hannon, Patrick R et al. (2017) Coordinated Regulation Among Progesterone, Prostaglandins, and EGF-Like Factors in Human Ovulatory Follicles. J Clin Endocrinol Metab 102:1971-1982
Kim, Soon Ok; Trau, Heidi A; Duffy, Diane M (2017) Vascular endothelial growth factors C and D may promote angiogenesis in the primate ovulatory follicle. Biol Reprod 96:389-400
Cacioppo, Joseph A; Koo, Yongbum; Lin, Po-Ching Patrick et al. (2016) Generation of an estrogen receptor beta-iCre knock-in mouse. Genesis 54:38-52
Cerny, Katheryn L; Ribeiro, Rosanne A C; Jeoung, Myoungkun et al. (2016) Estrogen Receptor Alpha (ESR1)-Dependent Regulation of the Mouse Oviductal Transcriptome. PLoS One 11:e0147685
Oakley, Oliver R; Kim, Kee Jun; Lin, Po-Ching et al. (2016) Estradiol Synthesis in Gut-Associated Lymphoid Tissue: Leukocyte Regulation by a Sexually Monomorphic System. Endocrinology 157:4579-4587
Trau, Heidi A; Brännström, Mats; Curry Jr, Thomas E et al. (2016) Prostaglandin E2 and vascular endothelial growth factor A mediate angiogenesis of human ovarian follicular endothelial cells. Hum Reprod 31:436-44
Wilson, Kalin; Park, Jiyeon; Curry Jr, Thomas E et al. (2016) Core Binding Factor-? Knockdown Alters Ovarian Gene Expression and Function in the Mouse. Mol Endocrinol 30:733-47
Kim, Soon Ok; Duffy, Diane M (2016) Mapping PTGERs to the Ovulatory Follicle: Regional Responses to the Ovulatory PGE2 Signal. Biol Reprod 95:33

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