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Binding of the sex steroids estradiol and progesterone to luteal tissue was studied in vitro in the presence and absence of added follicular fluid (FF) of cows. Bovine luteal tissue in the range of 2.0-10.0 mg of protein and granulosa cells were incubated at 37 degrees C for 2 hr with varying amounts of [3H]estradiol (4 nM-4 microM) or [3H]progesterone (300 nM-3 microM). Estrogen was found to bind in a dose-dependent manner to luteal tissue (2.0-4.0 mg of protein) and to granulosa cells (10.0 mg of protein) with a binding capacity of 7-10 fmol/100 mg of protein. Progesterone binding was also saturable and reversible in luteal tissue (2.0-10.0 mg of protein). The maximum binding capacity was 1,500 pmol/mg of protein and binding was not observed in tissues from older animals. Binding studies were also carried out in the absence and presence of FF (1.5 mg of protein), and in the absence of added FF in the presence of 10% molar excess of [3H]estradiol or [3H]progesterone, or specific antisera to estradiol or progesterone. The addition of FF to culture medium resulted in a significant dose-dependent decrease in the amount of hormone bound to the tissue at a given concentration. FF added at the higher concentration (1.5 mg of protein) decreased both the extent of binding and the total binding capacity. Specific antisera to estradiol and progesterone also decreased steroid-binding activity. The capacity of FF to displace steroid binding was found to be greatest for progesterone at concentrations between 300 and 1,500 pmol/ml. These results show that FF contains an inhibitor of steroid-binding to luteal tissue and that this inhibition is species-specific, competitive in nature, and correlates with the concentration of FF present in the culture medium.I saw A Feast for Crows, and even though I
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