FIGURE 3.

Forskolin, 8-bromo-cAMP, and estradiol effects on the TSHR transcript in the ovary. To evaluate the cAMP effects on TSHR expression, granulosa cells from 26-day-old rats were treated with graded doses of forskolin or 2 mm 8-bromo-cAMP (Br-cAMP) for 12 h (A). C, control. To test the estradiol effect, early antral follicles isolated from immature rats were incubated in the presence of 10⁻⁸ m estradiol for different intervals (B), and granulosa cells from 26-day-old rats were cultured in the presence of graded doses of estradiol for 12 h (C). After reverse transcription, the TSHR transcript levels were determined by real-time PCR and normalized using β-actin levels. Data were obtained from triplicate experiments and are shown as the means ± S.D. *, p < 0.05.