New clues to identify proteins correlated with Attractin

Summary In our previous study, the age-dependent testis vacuolisation and sperm dysfunction were found in Attractin (Atrn)-deficient mice, Atrnmg-3J, which is a null or nearly null allele. To explore the potential mechanism involved in these pathological changes, Attractin knock-down in mouse Sertoli cells TM4 (psiAtrn-TM4) was successfully established by stable RNA interference. The TM4 transfected by psiRNA-hH1 (psiRNA-TM4) was the control group, in which the expression of Atrn was not affected. The proteomic changes among the psiAtrn-TM4, primary cultures of Sertoli cells of Atrnmg-3J (mu-Sc) and control cells (psiRNA-TM4) were compared by two-dimensional gel electrophoresis. Fifteen differentially expressed protein spots of those cells were identified by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry and the NCBI proteins database. Except the decreased expression of superoxide dismutase (SOD), there were several novel proteins associated with Atrn function, including downregulated ATP synthase, peroxiredoxin 2 and upregulated caspase 6, ketohexokinase, etc., in psiAtrn-TM4 and mu-Sc. These data suggest that these differentially expressed proteins may be associated with the function of Atrn in Sertoli cells, thus providing a new clue to interpret the mechanism of testis degeneration in Atrn mutants.