Effects of glial cell line-derived neurotrophic factor on the proliferation and differentiation of PLZF and c-Kit of spermatogonial stem cells of rats in vitro

Abstract

Objective To investigate the effects of glial cell line-derived neurotrophic factor(GDNF)on the gene transcriptions and expressions of promyelocytic leukaemia zinc finger(PLZF)and c-Kit in spermatogonial stem cells(SSC).Methods SSC was cultured in DMEM plus different concentrations of GDNF(0,10,50,100 ng/ml).Quantitative reverse transcription PCR was used to detect the mRNA of PLZF and c-Kit.The expressions of PLZF and c-Kit protein were detected by Western blot assay.Results With the control group and 10ng/ml of GDNF group cell growth has no obvious influence,but with 50 ng/ml of GDNF group and 100 ng/ml of GDNF group cell growth was enhanced significantly.In the control group,the PLZF mRNA was 0.28±0.13 and c-Kit mRNA was 0.65±0.21.In the 10 ng/ml of GDNF group,PLZF mRNA was 0.27±0.14 and c-Kit tuRNA was 0.62±0.19.Compared with the control group,10 ng/ml of GDNF group had not influence on PLZF and c-Kit mRNA.In the 50 ng/ml of GDNF group PLZF mRNA was 0.64±0.28 and c-Kit mRNA was 0.34±0.15.Compared with the control group,50 ng/ml of GDNF enhanced the transcription of PLZF mRNA(P＜0.05),decreased the transeription of c-Kit mRNA (P＜0.05).In the 100 ng/ml of GDNF gmup,PLZF mRNA was 0.68±0.27 and c-Kit mRNA was 0.28±0.18.In the 100 ng/ml of GDNF group,there was no difference compared with 50 ng/ml of GDNF.In the control group,PLZF protein was 0.34±0.13 and c-Kit protein was 0.72±0.27.In 10 ng/ml of GDNF group,PLZF protein was 0.38±0.18 and c-Kit protein was 0.69±0.26.Compared with the control group,10 ng/ml of GDNF had no influence on PLZF and c-Kit protein.In 50 ng/ml of GDNF group,PLZF protein was 0.68±0.26 and c-Kit protein was 0.35±0.15.50 ng/ml of GDNF enhanced the expression of PLZF protein(P＜0.05),decreased the expression of c-Kit protein(P＜0.05).In 100 ng/ml of GDNF group PLZF protein was 0.70±0.27 and c-Kit protein Was 0.32±0.11,100 ng/ml of GDNF had no influence on PLZF and c-Kit protein compared with 50 ng/ml of GDNF.Conclusions Higher concentration of GDNF could enhance the transcription and expression of PLZF,and decrease the transcription and expression of c-Kit in SSC.GDNF has possible role to regulate and control proliferation and differentiation of SSC. Key words: Spermatogonia; Glial cell line-derived neurotrophic factor; Promyelocytic leukaemia zinc finger; Proto-oncogene proteins; c-kit