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The Preovulatory Increase in Ovarian Collagenase Activity in the Rat Is Independent of Prostaglandin Production^*

THOMAS E. CURRY, JR., MARTIN R. CLARK, DAVID D. DEAN, J. FREDERICK WOESSNER, JR. and WILLIAM J. LEMAIRE

The Reproductive Sciences and Endocrinology Laboratories, Departments of Obstetrics and Gynecology and Biochemistry, University of Miami School of Medicine Miami, Florida 33101

Address requests for reprints and all correspondence to: Thomas E. Curry, Jr., The Reproductive Sciences and Endocrinology Laboratories, D-5, Department of Obstetrics and Gynecology, P.O. Box 016960, University of Miami, Miami, Florida 33101.

Abstract

In the present study, we have examined the role of gonadotropins and prostaglandins in the preovulatory increase of ovarian collagenase activity in the rat. Whole ovaries of immature PMSG-primed rats (20 IU) were removed before and 8 h after the rats were treated with human (h) CG, Nembutal, and/or indomethacin. The ovaries were homogenized in a solution containing Triton X-100 (0.25%) and centrifuged. Collagenase was extracted by resuspending the pellets in buffer containing 100 mM CaCl₂, heating to 60 C for 6 min, and centrifuging. The supernatants were treated with dithiothreitol (2 mM) and iodoacetamide (5 mM) to inactivate collagenase inhibitors. Collagenase activity was measured as the percent digestion of ³H-type I collagen/100 µl aliquot of ovarian sample. At zero time (52 h after PMSG), ovarian collagenase activity was 4.2 ± 1.2% digestion (mean ± SEM, n = 3). In ovaries collected 8 h after the endogenous LH surge or 8 h after the administration of 10 IU hCG at time zero, collagenase activity rose to 19.6 ± 2.1 (n = 6) and 22.5 ± 1.7% digestion (n = 11), respectively. Indomethacin (1.5 mg/100 g BW) administered 30 min after hCG, produced no change in collagenase activity (24.8 ± 2.5% digestion, n = 7) although the expected increase in ovarian prostaglandin E after hCG treatment was blocked. When the endogenous LH surge was blocked with Nembutal (3 mg/100 g BW), collagenase activity in 8-h ovaries was 6.8 ± 1.1% digestion (n = 10). The Nembutal block of the preovulatory collagenase increase was overcome by administration of hCG (8-h ovarian enzyme activity = 22.7 ± 3.2% digestion, n = 8). These observations demonstrate that hCG stimulates ovarian collagenase activity and that this stimulation is not dependent on prostaglandin synthesis. (Endocrinology 118: 1823–1828, 1986)

Footnotes

^* This work was supported by NIH Grants HD-08747 (to W.J.L.) and HD-06773 (to J.F.W.) and Training Grant HD-07129.

Postdoctoral Fellow of the Arthritis Foundation.

Received September 17, 1985.

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