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Biology of Reproduction 62, 1335-1343 (2000)
© 2000 Society for the Study of Reproduction, Inc.


Articles

Intrafollicular Concentrations of Steroids and Steroidogenic Enzymes in Relation to Follicular Development in the Mare1

François Belina, Ghylène Goudeta, Guy Duchampa, and Nadine Gérard2,a

a INRA-Haras Nationaux, Reproduction Equine, P.R.M.D., 37380 Nouzilly, France

The objective of the present study was to determine the changes in follicular fluid steroid concentrations and in granulosa cell steroidogenic enzyme expression during the follicular phase, in relation to follicular size and physiological status in the mare.

Follicular fluid and follicular cells were recovered by ultrasound-guided follicular punctures either around the time of emergence of the dominant follicle, at the end of the dominant follicle growth, or at the preovulatory stage, after injection of gonadotropin to induce ovulation. Cellular relative amounts of steroidogenic acute regulatory protein (StAR), P450-side chain cleavage (P450scc), 3ß-hydroxysteroid dehydrogenase (3ßHSD), 17{alpha}-hydroxylase, and aromatase were assessed by semiquantitative Western blot and densitometry. Follicular fluid was assayed for cholesterol concentrations by colorimetric assay and for progesterone, testosterone, and estradiol-17ß concentrations by RIA.

Intrafollicular concentrations of progesterone and estradiol-17ß significantly increased in the dominant follicle during growth. After injection of gonadotropin, follicular maturation was characterized by a decrease in estradiol-17ß concentrations and a further increase in progesterone concentrations. Granulosa cells from dominant follicles had increased levels of StAR, P450scc, 3ßHSD, and aromatase during growth, but decreased levels during maturation. Levels of StAR, P450scc, 3ßHSD, and aromatase, as well as progesterone and estradiol-17ß, were lower in granulosa cells from subordinate than from dominant follicles. We did not observe a relationship between the steroidogenic activity of follicles and the capacity of their enclosed oocytes to complete meiosis in vitro.

First decision: 29 September 1999.

1 This work was supported by grants from the Institut National de la Recherche Agronomique (INRA, France) and the Haras Nationaux (France).

2 Correspondence. FAX: 33 2 47 42 77 43; gerard{at}tours.inra.fr




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