Puttabyatappa Muraly, Jacot Terry A, Al-Alem Linah F, Rosewell Katherine L, Duffy Diane M, Brännström Mats, Curry Thomas E
Department of Obstetrics and Gynecology, Chandler Medical Center, University of Kentucky, Lexington, Kentucky.
Department of Obstetrics and Gynecology, Eastern Virginia Medical School, Norfolk, Virginia.
Biol Reprod. 2014 Aug;91(2):34. doi: 10.1095/biolreprod.113.115717. Epub 2014 Jun 11.
An intrafollicular increase in proteolytic activity drives ovulatory events. Surprisingly, the periovulatory expression profile of the membrane-type matrix metalloproteinases (MT-MMPs), unique proteases anchored to the cell surface, has not been extensively examined. Expression profiles of the MT-MMPs were investigated in ovarian tissue from well-characterized rat and macaque periovulatory models and naturally cycling women across the periovulatory period. Among the six known MT-MMPs, mRNA expression of Mmp14, Mmp16, and Mmp25 was increased after human chorionic gonadotropin (hCG) administration in rats. In human granulosa cells, mRNA expression of MMP14 and MMP16 increased following hCG treatment. In contrast, mRNA levels of MMP16 and MMP25 in human theca cells were unchanged before ovulation but declined by the postovulatory stage. In macaque granulosa cells, hCG increased mRNA for MMP16 but not MMP14. Immunoblotting showed that protein levels of MMP14 and MMP16 in rats increased, similar to their mRNA expression. In macaque granulosa cells, only the active form of the MMP14 protein increased after hCG, unlike its mRNA or the proprotein. By immunohistochemistry, both MMP14 and MMP16 localized to the different ovarian cell types in rats and humans. Treatment with hCG resulted in intense immunoreactivity of MMP14 and MMP16 proteins in the granulosa and theca cells. The present study shows that MMP14 and MMP16 are increased by hCG administration in the ovulating follicle, demonstrating that these MMPs are conserved among rats, macaques, and humans. These findings suggest that MT-MMPs could have an important role in promoting ovulation and remodeling of the ovulated follicle into the corpus luteum.
卵泡内蛋白水解活性的增加驱动排卵事件。令人惊讶的是,膜型基质金属蛋白酶(MT-MMPs)这种锚定在细胞表面的独特蛋白酶在排卵前后的表达谱尚未得到广泛研究。我们在特征明确的大鼠和猕猴排卵模型以及自然周期女性的卵巢组织中,研究了整个排卵周期MT-MMPs的表达谱。在六种已知的MT-MMPs中,大鼠注射人绒毛膜促性腺激素(hCG)后,Mmp14、Mmp16和Mmp25的mRNA表达增加。在人颗粒细胞中,hCG处理后MMP14和MMP16的mRNA表达增加。相比之下,人卵泡膜细胞中MMP16和MMP25的mRNA水平在排卵前未发生变化,但在排卵后期下降。在猕猴颗粒细胞中,hCG增加了MMP16的mRNA,但未增加MMP14的mRNA。免疫印迹显示,大鼠中MMP14和MMP16的蛋白水平增加,与其mRNA表达情况相似。在猕猴颗粒细胞中,与mRNA或前体蛋白不同,hCG处理后只有MMP14蛋白的活性形式增加。通过免疫组织化学,MMP14和MMP16在大鼠和人类的不同卵巢细胞类型中均有定位。hCG处理导致颗粒细胞和卵泡膜细胞中MMP14和MMP16蛋白产生强烈的免疫反应性。本研究表明,排卵卵泡中hCG给药可使MMP14和MMP16增加,表明这些MMPs在大鼠、猕猴和人类中具有保守性。这些发现提示MT-MMPs在促进排卵以及将排卵后的卵泡重塑为黄体方面可能具有重要作用。