Influence mechanism of glial cell line-derived neurotrophic factor on the proliferation of spermatogonial stem cells

Abstract

Objective To investigate the molecular mechanisms of glial cell derived neurotrophic factor in promoting proliferation of spermatogonial stem cell. Methods RNAi expression vectors, targeted at GDNF, were constructed and transfected into SSCs from 5 to 7 days old mice.The SSCs with highest effectiveness of GDNF interfere was set as study group.And the SSCs without GDNF interfere was considered as control group. The ELISA method was used to compare the proliferative rate between study group and control group.Flow cytometry, RT-PCR were used to detect the expression of GDNF, RTKs, Fyn and FAK′s mRNA, and the apoptosis of SSCs. Results From 1 to 4 days after transinfection, the absorbable A value in study group was 0.45±0.02, 0.68±0.03, 1.12±0.03, 2.24±0.04, respectively.Meanwhile, the same item in control group was 0.46±0.03、0.73±0.02、1.32±0.05、1.15±0.06, respectively (P<0.05). There were significant different between experiment groups (25.43±1.91)% and control group (5.61±0.16)% in the apoptosis rates of SSCs (P<0.05). Significant differences were noted between experimental group and control group(P<0.05). The mRNA expression rates of GDNF was (12.32±1.22)% in study group and (54.25±1.34)% in control group (P<0.01). The mRNA expression rates of RTKs and Fyn and FAK in study group and control group were (16.24±1.35)% vs (45.35±1.37)%, (18.32±1.34)% vs (38.37±1.55)%, (20.04±1.65)% vs (43.27±1.28)%, respectively (P<0.05). Conclusions The glial cell line derived neurotrophic factor was important in course of SSCs′ proliferation, which may up-regulating the expression of RTKs, Fyn and FAK. Key words: Spermatogonial stem cells; Glial cell line derived neurotrophic factor; Molecular mechanism