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Molecular Biology and Functional Morphology Divisions, Women’s Health Research Institute, Wyeth-Ayerst Research, Radnor, Pennsylvania 19087
Address all correspondence and requests for reprints to: Dr. Susan L Fitzpatrick, Women’s Health Research Institute, Wyeth-Ayerst Research, 145 King of Prussia Road, Radnor, Pennsylvania 19087. E-mail: fitzpas2{at}war.wyeth.com
Estrogen is an essential hormone for the LH surge and ovulation. The
primary source of estrogen is from ovarian granulosa cells and in rats,
estrogen, in turn, increases granulosa cell number and enhances
FSH-stimulated gene expression in these cells. Thus, rat granulosa
cells both respond to and synthesize estrogen. To further elucidate the
mechanisms mediating the actions of estrogen in granulosa cells, we
have identified and characterized the estrogen receptor-ß (ER-ß)
subtype in rodent granulosa cells. ER-ß protein was localized to the
nuclei of rat granulosa cells in preantral and antral follicles by
immunocytochemistry, coincident with the location of ER-ß messenger
RNA (mRNA). Immunoprecipitation and Western blot analysis using ER-ß
specific antisera demonstrated a protein of approximately 60 kDa in
granulosa cells prepared from PMSG-primed immature mice and
estrogen-treated immature rats. Extracts from granulosa cells
specifically bound an estrogen response element and the complex was
recognized by antisera to ER-ß. A synthetic steroid estrogen
radioligand,
[125I]-17
-iodovinyl-11ß-methoxyestradiol
([125I]-VME2), bound to cytosolic granulosa cell
preparations with high affinity (estimated KD value of
401 ± 83 pM, and Bmax value of 102
± 9 fmol/mg protein). ER-ß protein levels rapidly declined following
hCG treatment consistent with the reported decrease in binding activity
and ER-ß mRNA levels by high levels of gonadotropins. Overall, we
have demonstrated that 1) ER-ß protein is the dominant estrogen
receptor subtype present in rodent granulosa cells, 2) this receptor is
functional, and 3) it is regulated by ovulatory doses of gonadotropins.
Thus, ER-ß is likely to be a mediator of estrogen action in rodent
granulosa cells during follicular development.
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