Cabrera Rafael A, Dozier Brandy L, Duffy Diane M
The Jones Institute for Reproductive Medicine, Department of Obstetrics and Gynecology, Eastern Virginia Medical School, Norfolk, Virginia, USA.
Fertil Steril. 2006 Oct;86(4 Suppl):1088-96. doi: 10.1016/j.fertnstert.2006.03.022. Epub 2006 Sep 7.
To determine whether hCG regulates the expression of prostaglandin (PG) synthesis enzymes and the production of PGs by normal monkey ovarian surface epithelium (OSE).
Experimental animal study.
Research laboratory.
ANIMAL(S): Adult cynomolgus macaques.
INTERVENTION(S): Monkeys received exogenous gonadotropins to stimulate multiple follicular development. Ovarian surface epithelium cells and whole ovaries were obtained before (0 hours) and 36 hours after an ovulatory dose of hCG.
MAIN OUTCOME MEASURE(S): Ovarian surface epithelium expression of prostaglandin-endoperoxide synthase 1 (PTGS1) and PTGS2 proteins was determined by immunocytochemistry. Prostaglandin synthesis enzyme messenger RNA (mRNA) levels were determined by RT-PCR. Prostaglandin E2 and PGF2alpha production was assessed by enzyme immunoassays.
RESULT(S): Ovarian surface epithelium maintained in long-term culture expressed mRNA and protein for PTGS1 and PTGS2 (n = 6); inhibition of PTGS1, but not PTGS2, reduced PGE2 synthesis (n = 3). Prostaglandin-endoperoxide synthase 1 was present in OSE of ovarian tissue sections obtained 0 (n = 4) and 36 (n = 3) hours after hCG; PTGS2 was not detected. Ovarian surface epithelium collected 0 (n = 3) and 36 (n = 4) hours after hCG expressed mRNAs for PTGS1, PTGS2, and three PGE synthases; the ratio of PTGS2 to PTGS1 increased in response to hCG exposure.
CONCLUSION(S): Monkey OSE expresses mRNA for PTGS1, PTGS2, and all PGE synthases and produces PGE2 both before and 36 hours after hCG. Detection of PTGS1, but not PTGS2, protein in OSE in vivo supports the hypothesis that PTGS1 is the enzyme responsible for PGE2 production by primate OSE in vivo.
确定人绒毛膜促性腺激素(hCG)是否调节正常猴卵巢表面上皮(OSE)中前列腺素(PG)合成酶的表达及PG的产生。
实验动物研究。
研究实验室。
成年食蟹猴。
给猴子注射外源性促性腺激素以刺激多个卵泡发育。在注射排卵剂量的hCG前(0小时)和注射后36小时获取卵巢表面上皮细胞和整个卵巢。
通过免疫细胞化学法测定卵巢表面上皮中前列腺素内过氧化物合酶1(PTGS1)和PTGS2蛋白的表达。通过逆转录聚合酶链反应(RT-PCR)测定前列腺素合成酶信使核糖核酸(mRNA)水平。通过酶免疫测定法评估前列腺素E2和前列腺素F2α的产生。
长期培养的卵巢表面上皮表达PTGS1和PTGS2的mRNA和蛋白(n = 6);抑制PTGS1而非PTGS2可降低前列腺素E2的合成(n = 3)。在注射hCG后0小时(n = 4)和36小时(n = 3)获取的卵巢组织切片的OSE中存在前列腺素内过氧化物合酶1;未检测到PTGS2。在注射hCG后0小时(n = 3)和36小时(n = 4)收集的卵巢表面上皮表达PTGS1、PTGS2和三种前列腺素E合成酶的mRNA;暴露于hCG后,PTGS2与PTGS1的比值增加。
猴OSE表达PTGS1、PTGS2和所有前列腺素E合成酶的mRNA,在hCG注射前及注射后36小时均产生前列腺素E2。在体内OSE中检测到PTGS1蛋白而非PTGS2蛋白,支持了PTGS1是灵长类动物体内OSE产生前列腺素E2的负责酶这一假说。