Gelatinase and proteoglycanase activity during the periovulatory period in the rat.
Gelatinase and proteoglycanase are metalloproteinases that govern extracellular matrix remodeling. In the present study, immature rats were primed with eCG (20 IU) and hCG (10 IU). Ovarian gelatinase and proteoglycanase activity were determined at the time of hCG administration (0 h) as well as 4, 8, and 12 h later. Gelatinase and proteoglycanase were extracted by homogenization in Triton and by heating (i.e., heat extraction). An aliquot of the heat extract was reduced and alkylated to destroy metalloproteinase inhibitors. Heat extracts not reduced and alkylated showed low levels of gelatinase and proteoglycanase activity that did not change at the different time points. However, with reduction and alkylation, gelatinolysis increased approximately 4-fold (p less than 0.05) at 4 h, 8 h, and 12 h after hCG priming. Proteoglycanase activity increased approximately 2-fold (p less than 0.05) between 0 and 8 h and declined at 12 h after hCG. The ovarian gelatinolytic activity was due to a metalloproteinase as demonstrated by the inhibition of enzyme activity by phenanthroline and EDTA (97.1 +/- 0.7% and 97.4 +/- 0.6% inhibition respectively). Proteoglycanase activity was not inhibited by phenanthroline (11.5 +/- 3.5%), suggesting that the enzyme activity was not specifically a metal-dependent enzyme. Gelatin gel zymography of the ovarian extracts demonstrated four predominant and distinct gelatin-degrading enzymes of 78, 72, 66, and 62 kDa, similar to the size of gelatinase. The present findings demonstrate a periovulatory increase in ovarian gelatinolytic and proteglycanase activity that may play a pivotal role in connective tissue remodeling associated with ovulation.
Curry
T E
TE
Department of Obstetrics & Gynecology, University of Kentucky, Lexington 40536.
Mann
J S
JS
Huang
M H
MH
Keeble
S C
SC
D000818:Animals; D010450:Endopeptidases / Q000378:metabolism*; D005260:Female; D018093:Gelatinases; D006107:Granulosa Cells / Q000201:enzymology; D066298:In Vitro Techniques; D008666:Metalloendopeptidases; D010053:Ovary / Q000201:enzymology; D010060:Ovulation / Q000502:physiology*; D010434:Pepsin A / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains
D011480:Protease Inhibitors; D010450:Endopeptidases; C022474:proteoglycan-degrading metalloendopeptidases; D010434:Pepsin A; D018093:Gelatinases; D008666:Metalloendopeptidases