Abstract
Purpose :
Sox2 is a neuronal stem cell-associated transcription factor, regulates neural development in the vertebrates, and maintains neural stem cell identity in the central nervous system. This study is to identify a functional role for Sox2 during bipolar cell differentiation in the developmental mouse retina.
Methods :
Injection and electroporation in P0 CD1 mouse retina. 1 μl of shRNA to knockdown Sox2 (pSico-shSOX2+IRES+GFP) (3 μg/μl) or vehicle only was injected into the subretinal space, using a Microliter Syringe (Hamilton). Immediately following injection, electric pulses (100 V; five 50 ms pulses with 950 ms pause intervals) were applied through tweezer-type electrodes using the pulse generator ECM 830 (BTX). Scotopic ERG was performed to check the retina neuron function at P12, P30 and P60. IOP was also monitored every month. Transfected retinas were collected for analysis; Immunohistochemistry cell staining with type-specific markers were performed to identify retinal neurons number change. RNA-seq was performed to identify RNA changes of the retina when sox2 is inhibited.
Results :
At P12, P30, P60, inhibition of Sox2 leads to a significant unregulated bipolar cells number and a decrease number of amacrine cells in the retinas. However, very surprisingly, the scotopic ERG b-wave amplitude significantly decreased in sox2i-treated retinas, indicate the gained bipolar cells either has no function, or the overall retina bipolar cells function decreased as well. IOP has no change during the whole process. RNA-seq analysis of developmental stage-specific retinal transcriptomes showed that Vsx2, Isl1, Otx2 etc. which were transcription factors of bipolar cells had a high level of expression in P12.
Conclusions :
Sox2 control the RBCs differentiation and modulate function of RBCs in the mouse developmental stage.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.