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Biology of Reproduction 65, 726-732 (2001)
© 2001 Society for the Study of Reproduction, Inc.


Regular Article

Role of Gelatinase on Follicular Atresia in the Bovine Ovary1

M.A.M. Yahia Khandokera,b, Kei Imaia, Toru Takahashia, and Kazuyoshi Hashizume2,a

a National Institute of Agrobiological Sciences, Kukizaki, Ibaraki ken 305-0901, Japan b Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

ABSTRACT

Follicular atresia, like follicular growth and ovulation, is characterized by excessive tissue remodeling. It is hypothesized that probably one of the tissue-remodeling enzymes, such as the gelatinases, could be playing an important role in this process. The present study was undertaken to determine the role of gelatinase on follicular atresia in the cow. Follicles of 2–6 mm in diameter were dissected from ovaries, and follicular fluid was categorized according to the morphological appearance of the cumulus-oocyte complexes. Gelatinase activity within the follicular fluid was analyzed by gelatin zymography, and film in situ zymography was employed in order to localize gelatinase. TUNEL was performed on cryosectioned ovaries to understand follicular health. The concentrations of steroids in follicular fluid were also measured by solid phase fluoroimmunoassay. ProMMP-2 was detected in all normal and atretic categories of follicular fluid. The active form of MMP-2 and an additional band of proMMP-9 were detected only in atretic follicular fluid. Gelatinase activity was recorded in both granulosa cells (GCs) and theca cells (TCs) but were found in comparatively higher numbers in those follicles that exhibited a thinned and partially detached granulosa layer. TUNEL confirmed that apoptosis had commenced in the GCs of follicles of the latter category. The estradiol-17ß (E2):progesterone (P4) ratio was found to be significantly lower in atretic follicles than in normal follicles. These results suggest a plausible role for gelatinase in follicular health, especially the active form of MMP-2 and proMMP-9, and that bovine follicular fluid may be a key indicator of atresia.

FOOTNOTES

First decision: 5 December 2000.

1 M.A.M.Y.K. was a recipient of a Science and Technology Agency (STA) fellowship from the Japan Science and Technology Agency (JST) to carry out research at the Laboratory of Reproductive Endocrinology, National Institute of Animal Industry, Tsukuba, Ibaraki, Japan.

2 Correspondence: Kazuyoshi Hashizume, Laboratory of Reproductive Biology and Technology, National Institute of Agrobiological Sciences, Kukizaki Campus, 2 Ikenodai, Kukizaki, Ibaraki 305-0901, Japan. FAX: 81 298 38 8606; kazuha{at}affrc.go.jp




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