Localization of phosphorylated SMAD proteins in granulosa cells, oocytes and oviduct of female mice.

SMAD signaling is essential for follicular development. The distribution of activated (phosphorylated) SMADs during folliculogenesis has not been described in detail. The present results indicate that oocytes from preantral and antral follicles contain the mRNA for the receptor regulated Smads (-1, -2, -3, -5 and -9), Smad4 (co-SMAD), and Smad6 and Smad7 (inhibitory SMADs). Levels of Smad5, Smad6 and Smad7 mRNA decreased in fully-grown oocytes compared to growing oocytes. Immunostaining for pSMAD1/5/9, pSMAD2 and pSMAD3 was evident in oocytes from primordial, primary, secondary and antral follicles. We also observed substantial staining for pSMADs in intact fully-grown oocytes from antral follicles. In granulosa cells, immunostaining for both pSMAD2/3 and pSMAD1/5/9 was apparent in preantral granulosa cells from primordial to secondary follicles. In antral follicles, immunostaining for pSMADs became more intense in the cumulus cells surrounding the oocyte. Immunostaining for pSMAD2 and pSMAD1/5/9 was also apparent in the epithelium lining of the oviduct. Clearly, oocytes, preantral granulosa cells, cumulus cells and the epithelium of the oviduct are major targets of SMAD-mediated pathways in female reproductive tissues. The finding that pSMAD pathways are active in oocytes raises new questions regarding the role of TGF-beta superfamily members in directly promoting oocyte development. The extent to which defects in pSMAD signaling in oocytes or the oviduct contribute to infertility in humans or animals remains an open question.