Fig. 8.
Time-dependent changes of Ctgf and Adamts1 mRNA expression in granulosa cells of Ctgf cKO and Pgr mutant mice during the preovulatory period. Real-time qPCR analysis of granulosa cell mRNA from 6- to 8-month-old adult control and Ctgf cKO (Pgrcre/+) female mice and from 3-wk-old Pgr+/− and Pgr−/− female mice treated with PMSG for 46 h before the stimulation of ovulation by hCG. Granulosa cells were collected at 0 h (only 46 h PMSG), 6 h, and 11 h after administration of hCG from ovaries of five independent mice. Total RNA was isolated and converted to cDNA. Ctgf, Adamts1, and other progesterone-regulated genes-mRNA levels were analyzed over time. A, Ctgf is induced at 6 h after hCG stimulation, followed by a continuing declines at 6 h and 11 h after hCG stimulation in preovulatory granulosa cells from control mice. In contrast, the expression of Ctgf is significantly down-regulated in Ctgf cKO (Pgrcre/+) female mice compared with the control expression. B, Adamts1 is induced in a time-dependent manner after hCG stimulation in preovulatory granulosa cells from control mice. However, the expression of Adamts1 is significantly down-regulated in Ctgf cKO (Pgrcre/+) female mice, according with the reduction of Ctgf expression. C–E, The expression of Edn2, Il6, and Prkg2 was induced after hCG stimulation and unchanged between control and Ctgf cKO (Pgrcre/+) female mice. The control values were set to equal 1. *, P < 0.05; **, P < 0.01; ***, P < 0.001 (vs. control female mice). F, PGR regulates the induction of Ctgf mRNA during ovulation. Pgr+/− and Pgr−/ − female mice (age 3 wk) were treated with PMSG for 46 h before the stimulation of ovulation by hCG. Granulosa cells were collected at 0 h (only 46 h PMSG) and 6 h after administration of hCG from Pgr+/− and Pgr−/− ovaries (n = 3–5). Total RNA was isolated and real-time PCR was used to measure Ctgf mRNA levels. The expression of Ctgf mRNA in Pgr+/− granulosa cells at 0 h was set to equal 1. **, P < 0.01 (vs. Pgr+/− female mice). RQ, relative quantity.