Hormonal regulation of tissue inhibitors of metalloproteinases during follicular development in the rat ovary

Abstract

Tissue inhibitors of metalloproteinases (TIMPs) are membres of a multigene family of proteinase inhibitors that regulate the activity of metalloproteinases. To test the hypothesis that TIMPs regulate connective tissue remodeling during follicular development, rats were injected with PMSG (20 IU, sc), and ovaries and serum were collected at the time of pregnant mare serum gonadotropin at the time of pregnant mare serum gonadotropin (PMSG) administration (0 h) and at 6, 12, 24, 36, and 48 h later for analysis of TIMP expression, metalloproteinase inhibitor activity, and steroidogenesis. Serum estradiol levels increased from 20.9 pg/mL at 0 h to 461 pg/mL at 48 h. Northern analysis was performed for analysis of TIMP-1, TIMP-2, and TIMP-3 expression (N = 4). For TIMP-1, PMSG stimulated a 2.4- to 2.5-fold increase in TIMP-1 mRNA at 6 and 12 h compared to ovaries collected at the time of PMSG administration (i.e., 0 h control). TIMP-1 mRNA returned to control levels within 24 h and remained unchanged through 48 h. In contrast to TIMP-1, TIMP-3 mRNA decreased by approx 2.5-fold at 6 h following PMSG administration, and expression remained decreased through 48 h. For TIMP-2, the expression of the 3.5-kb transcript decreased at 24 h after PMSG, whereas expression of the 1 kb transcript was unchanged. There was no change in metalloproteinase inhibitor activity in whole ovarian extracts between 0 and 36 h. However, there was an increase in inhibitor activity at 48 h. These findings are the first demonstration of hormonal regulation of TIMPs during the follicular phase. The differential regulation of the TIMPs by gonadotropins, for example, an increase in TIMP-1 and a concomitant decrease in TIMP-3 expression, may reflect different roles, sites of action, or enzyme specificity for the inhibitors as the follicle grows.