Regulation of transforming growth factor alpha on glutamate transpoter of retinal M(u)ller cells in mice / 中华眼底病杂志

ABSTRACT

Objective To observe the regulation effect of transforming growth factor alpha(TGFα) on expression of glutamate transporter(GLAST)and ingestion activity of retinal M(u)ller cells in mice.Methods To take the retinal tissue of Kunming mouse at postnatal 7～10 day.and then cultured M(u)ller cells according to literature.The 3～4 generation cultured cells of the same primary cell were divided into two groups at random:①TGFα group:maintained in different concentrations of TGFα as 50,75,125 and 150 ng/ml,3 holes in each concentration;②Control group:cultured by Eagle culture medium which improved from Dulbeccon and contained 20%fetal calf serum.The influence of different concentrations TGFα on GLAST activity in M(u)ller cells were observed by L-3H-glutamate uptake detection;the expression of GLAST mRNA in M(u)ller cells was determined by RT-PCR;the expression of GLAST protein was detected with immunocytochemical staining.Results With the increase of TGFα concentration.both L-3H-glutamate uptake and GLAST mRNA expression were increased.The L-3H-glutamate accumulation had got to the maximum uptake at concentration of 125 ng/ml,which was 266% of that in control group,meanwhile,the expressions of GLAST mRNA also got to the maximum as 4 times of control group.Immunocytochemical staining indicated that the effect of 125ng/ml TGFα on expression of GLAST protein was higher than that in the control group,the differences between two groups were statistically significant(P＜0.05).Conclusion TGF-α can increase GLAST activity through up-regulating the expression of GLAST mRNA and protein.