Abstract
Purpose: :
To identify the differential expressed proteins induced by stroboscopic light in form-deprived C57 mice retina via functional proteomics approach.
Methods: :
Forty-eight mice, aged 27-29days, were randomly divided into three groups: only form-deprived group (FD+/SL-), form-deprivation & stroboscopic illumination group (FD+/SL-) and control group (each N=16). The right eyes of mice in FD+/SL- group and FD+/SL+ group were attached with diffusers to impose form-deprivation, while the contralateral eyes were opened to clear vision as self-control. The mice in FD+/SL- group and control group were kept on normal room light, however, the mice in FD+/SL+ group were exposed to 15HZ stroboscopic light only. At the end of the two-week treatment period, the retinas were prepared for two-dimension gel electrophoresis (2D) and the gels were stained by Coomassie Brilliant Blue R-250. The ImageMaster 2D Platinum software was used to analysis the protein spots in gels. The differential expressed protein spots were identified by MALDI-TOF Mass Spectrometry (MS). Western blot was finally reverified the results of 2D.
Results: :
(1) the results of 2D-MS: five proteins, later successfully identified as alpha-A-crystalline, crystalline beta A1, alpha-A-crystalline, crystalline beta A2 and crystalline beta A1 by MS respectively, were found down-regulated in FD+/SL+ retinas compared with FD+/SL- retinas. (2) the results of western blot: the expression level of crystalline alpha-A in FD+/SL+ retinas is significantly lower than that in FD+/SL- retinas, and even lower than that in C retinas; surprisingly, the expression level of crystalline alpha-A in FD+/SL+ retinas and FD+/SL- retinas is also significantly lower than their contralateter self-control retinas.
Conclusions: :
Under the conditions of our experiments, stroboscopic illumination and form-deprivation for two weeks could induce significantly differential expression of some members of crystalline family in mice retina. Recent researches provide evidence that some members in crystalline family act as hot shock proteins and some possess autophosphorylation activity. Therefore, we could expect that some members in crystalline family might involve in ocular growth regulation. Future research is needed for identify the ocular growth regulation network including crystalline family.This work was supported by a grant from the National Natural Science Foundation of China (30572007, 30772389) and SRFDP (20050558075).
Keywords: myopia • retina • proteomics