| ObjectiveIn early studies, we have shown that the expression of proto-oncogene c-erbB2 was found during primordial follicles growth and using RNA interference method confirmed c-erbB2 played an important role in this progress. In this experiment, we investigated the upstream or downstream relationship of proto-oncogene c-erbB2 with PKC and MAPK signaling pathway in the initiation of primordial follicle growth in rats.Methods1. RT-PCR and western blot analysis were performed to measure the expressions of c-erbB2mRNA as well as ErbB2,MAPK and PKC protein after the neonatal rat ovaries were transfected with c-erbB2siRNA.2. RT-PCR and western blot analysis were performed to measure the expressions of c-erbB2mRNA and ErbB2 after neonatal rat ovaries in vitro were cultured with PD98059 and calphostin for 8d.3. The morphous and quantity of every stage follicles of ovaries cultured for 8d were observed by histological section stained with hematoxylin after ovaries were transfected with c-erbB2siRNA and cultured with PD98059 and calphostin.Results1. After siRNA transfected, the levels of c-erbB2mRNA expressed in cultured ovaries were decreased significantly(P<0.05)by RT-PCR and the levels of ErbB2,MAPK and PKC protein also were decreased remarkedly by western blot analysis(P<0.05).2. After PD98059 and calphostin effect, the levels of c-erbB2mRNA expressed in cultured ovaries did not change significantly (P>0.05)and the levels of ErbB2,MAPK and PKC protein also have no change significantly(P>0.05).3. After siRNA transfected, the initiation of primordial follicles were inhibited remarkably (P<0.01); After PD98059 and calphostin effect, the initiation of primordial follicles delayed obviously (P<0.05). Conclusion1. Proto-oncogene c-erbB2 play an important role on the initiation of primordial follicles growth through MAPK and PKC signal pathway.2. Proto-oncogene c-erbB2 maybe the upstream of PKC and MAPK signalling pathway in the regulation of primordial follicles growth.
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