The present studies investigated the hypothesis that TGFbeta plays a role in mediating LH/hCG-induced maturation, ovulation and/or luteinization of follicles in the pig. In Experiment 1, the temporal and spatial gene expression patterns of TGFbeta signaling components were examined in pig follicles which had been induced to ovulate and luteinize in vivo by hCG treatment, or by the LH-surge. Pre-pubertal pigs were injected with PG-600 followed by hCG, and ovaries were collected surgically at 0, 1, 12, 24 and 48h post-hCG. Post-ovulatory follicles were also collected from cycling gilts on Day 4 (D4) of the estrous cycle. Pre- and post-ovulatory follicles were used for the measurement of mRNA (PCR) and protein (Western blots) abundance and for protein localization by immunohistochemistry (IHC). Steady state amounts of mRNA for TGFbeta3 and TGFbetaR2 were increased (P<0.05, as compared to 0h) at 12h and on D4, respectively, while TGFbeta2 protein showed a tendency to increase on D4. TGFbeta signaling components did not change significantly. By IHC, the localization of TGFbeta components was as follows: pre-ovulatory follicles; TGFbeta1 - granulosa cells (GC), TGFbeta2 - theca cells (TC), TGFbetaR1 and 2 - GC and TC: post-ovulatory follicles; TGFbeta1 and 2 and TGFbetaR1 and 2 - luteinizing TC and GC. In Experiment 2, TGFbeta1 (1-100ng/ml) alone had no significant effect on progesterone (P4) secretion by pig GC in culture. Furthermore, while LH+IGF-1 (positive control) stimulated P4 approximately 10-fold, TGFbeta at 10 and 100ng/ml added in combination with LH+IGF-1, had no effect on P4 accumulation. In conclusion, data from the present study on temporal and spatial patterns of expression of the TGFbeta-system suggest that TGFbeta may play a role in the overall process of luteinization, but it appears not to influence steroidogenesis in luteinizing pig follicles.
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