Boerboom D, Russell D L, Richards J S, Sirois J
Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, CP 5000, Saint-Hyacinthe, Québec, Canada J2S 7C6.
J Mol Endocrinol. 2003 Dec;31(3):473-85. doi: 10.1677/jme.0.0310473.
One member of a new family of metalloproteinases, a disintegrin and metalloproteinase with thrombospondin-like motifs-1 (ADAMTS-1), has been found to be expressed and hormonally induced in granulosa cells of ovulating rodent follicles. Furthermore, the targeted disruption of the ADAMTS-1 gene resulted in ovarian defects associated with severely impaired fertility. While these data demonstrate the importance of ADAMTS-1 in rodent ovarian physiology, the potential role of ADAMTS-1 in the ovulatory process of monoovulatory species remains unknown. The objectives of this study were to clone the equine ADAMTS-1 primary transcript and to study its regulation during human chorionic gonadotropin (hCG)-induced ovulation. A 3573 bp follicular cDNA library clone was isolated and found to encode a nearly complete, highly conserved ADAMTS-1 homologue. Real-time RT-PCR analysis detected this transcript in diverse tIssues, including previously unreported sites of ADAMTS-1 expression such as the male reproductive tract, the follicular theca interna and the mature corpus luteum. The tIssue distribution of the progesterone receptor (PR), a known regulator of ADAMTS-1 expression in rodent preovulatory follicles, was found to overlap that of ADAMTS-1 in some tIssues. A study of the regulation of follicular ADAMTS-1 and PR mRNAs during the hCG-induced ovulatory process revealed distinct patterns of regulation in granulosa cells and in theca interna. In granulosa cells, ADAMTS-1 mRNA was found to be induced at 12 h post-hCG (P<0.05), followed by a return to basal levels by 30 h and a re-increase at 33-39 h (P<0.05). A concomitant increase in PR mRNA (P<0.05) was observed at 12 h post-hCG. In theca interna, abundant ADAMTS-1 mRNA was detected at all timepoints, and levels increased transiently at 33 h post-hCG (P<0.05), whereas no significant change was observed in PR mRNA. Together, these data demonstrate for the first time the hormonally regulated ovarian expression of ADAMTS-1 in a monoovulatory species, and identify a novel biphasic regulation of ADAMTS-1 in granulosa cells and a regulated expression in theca interna that were not previously observed in rodents.
金属蛋白酶新家族的一个成员，即含血小板反应蛋白基序的解聚素和金属蛋白酶-1（ADAMTS-1），已被发现在排卵的啮齿动物卵泡的颗粒细胞中表达且受激素诱导。此外，ADAMTS-1基因的靶向破坏导致与生育力严重受损相关的卵巢缺陷。虽然这些数据证明了ADAMTS-1在啮齿动物卵巢生理学中的重要性，但ADAMTS-1在单排卵物种排卵过程中的潜在作用仍然未知。本研究的目的是克隆马ADAMTS-1初级转录本，并研究其在人绒毛膜促性腺激素（hCG）诱导排卵过程中的调控。分离出一个3573 bp的卵泡cDNA文库克隆，发现其编码一个近乎完整、高度保守的ADAMTS-1同源物。实时RT-PCR分析在多种组织中检测到该转录本，包括ADAMTS-1表达的先前未报道的部位，如雄性生殖道、卵泡内膜和成熟黄体。已知在啮齿动物排卵前卵泡中ADAMTS-1表达的调节因子孕酮受体（PR）的组织分布，在某些组织中与ADAMTS-1的组织分布重叠。对hCG诱导排卵过程中卵泡ADAMTS-1和PR mRNA调控的研究揭示了颗粒细胞和内膜中不同的调控模式。在颗粒细胞中，发现ADAMTS-1 mRNA在hCG注射后12小时被诱导（P<0.05），随后在30小时恢复到基础水平，并在33-39小时再次增加（P<0.05）。在hCG注射后12小时观察到PR mRNA同时增加（P<0.05）。在内膜中，在所有时间点都检测到丰富的ADAMTS-1 mRNA，并且在hCG注射后33小时水平短暂增加（P<0.05），而PR mRNA未观察到显著变化。总之，这些数据首次证明了单排卵物种中ADAMTS-1在卵巢中的激素调节表达，并确定了颗粒细胞中ADAMTS-1一种新的双相调节以及内膜中的调节表达，这在啮齿动物中以前未观察到。
