Prostaglandin-endoperoxide synthase (PTGS1 and PTGS2) expression and prostaglandin production by normal monkey ovarian surface epithelium  Rafael A. Cabrera, M.D., Brandy L. Dozier, B.S., Diane M. Duffy, Ph.D.  Fertility and Sterility  Volume 86, Issue 4, Pages 1088-1096 (October 2006) DOI: 10.1016/j.fertnstert.2006.03.022 Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 Immunocytochemical detection of PTGS1 and PTGS2 in monkey ovarian surface epithelium (OSE). (A) Micrograph of a primary epithelial culture after 24 hours of incubation. (B) Confluent monolayer of cobblestone epithelial cells after first passage. (C) Cultured OSE expressed the epithelial-associated intermediate filament cytokeratin. Both PTGS1 (D) and PTGS2 (E) immunostaining consistently were present in long-term–cultured OSE cells. Omission of the primary antibody yielded no staining of the OSE cells (F). In a monkey ovary obtained after controlled ovarian stimulation and 36 hours of exposure to hCG, dark cytoplasmic precipitate representing PTGS1 immunostaining was present in the OSE (G) but not in granulosa or stromal cells (G, inset); PTGS2 immunostaining consistently was seen in the granulosa cells (H, inset) but was not evident in the OSE (H). Omission of the primary antibody yielded no staining of the granulosa cells or the surface epithelium (I). In G, st = ovarian stroma, gc = granulosa cells, and an = follicle antrum. Orientation of images is similar for panels G–I. Bar in F = 10 μm for panels A and B and = 20 μm for panels C–F. Bar in I = 25 μm for panels G–I. Cabrera. Prostaglandin production by monkey OSE. Fertil Steril 2006. Fertility and Sterility 2006 86, 1088-1096DOI: (10.1016/j.fertnstert.2006.03.022) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 2 Prostaglandin E2 levels in the presence of PTGS inhibitors. Long-term cultures of OSE (n = 3 animals) were treated with vehicle, the PTGS1-selective inhibitor SC-560, or the PTGS2-selective inhibitor NS-398 for 24 hours. Media PGE2 levels were determined by enzyme immunoassay. Prostaglandin E2 levels are expressed as a percentage of vehicle levels for each animal. Prostaglandin E2 production by monkey OSE varied between animals; the actual PGE2 level in vehicle wells was 33.7 ± 6.6 pg/mL. SC-560 reduced media PGE2 in a dose-dependent fashion; a>b>cP<.05. No differences were detected between vehicle and NS-398-treated cells. Data are expressed as mean + SEM. Cabrera. Prostaglandin production by monkey OSE. Fertil Steril 2006. Fertility and Sterility 2006 86, 1088-1096DOI: (10.1016/j.fertnstert.2006.03.022) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 3 Prostaglandin-endoperoxide synthase 1 and PTGS2 mRNA in monkey OSE before (0 hours) and 36 hours after hCG administration. Data represent the ratio of PTGS2 mRNA to PTGS1 mRNA and are expressed as mean + SEM (n = 3 for 0 hours and n = 4 for 36 hours). a<b, P<.05. Cabrera. Prostaglandin production by monkey OSE. Fertil Steril 2006. Fertility and Sterility 2006 86, 1088-1096DOI: (10.1016/j.fertnstert.2006.03.022) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions