Abstract
Abstract: :
Purpose: Embryonic stem cells can be induced to differentiate into many specialized cell types under the adequate cell culture conditions. The objective of this study is to establish a comparison between mouse embryo and mouse Embryonic Stem Cells (ESC) retinal genes. We analysed neuroectodermal genes, neural retinal precursor genes, transcription factors and protein genes related to photoreceptors. Methods: For in vivo analysis the OF–1 wild–type mouse strain was selected. Eyes from E9.5 to E18.5 embryos, from post–natal mice and from adults were used. ES–D3 embryonic stem cells were grown as embryoid bodies under spontaneous differentiation conditions for 30 days. RNA from both, the mouse eyes and the embryoid bodies at different days on culture, were obtained and analysed by RT–PCR. Pax6, otx2, neurofilament, notch1, Chx10, TrB2, Crx, Nrl, rhodopsin kinase, IRBP, Pdeb, rod arrestin, rod and cone transducin, rhodopsin, s–opsin, peripherin among other genes were studied. Results: We obtained the expression patterns of these genes for the two different sequences: mouse and embryoid bodies development. The mouse genetic sequence served us to have a positive control about the expression of the examined genes in order to establish the comparison of what happens in vivo and in vitro with the ESC. The embryoid bodies sequences helped us to know which of the examined genes are expressed and which are not in the embryoid bodies. Moreover, we found similarities and differences in the expression of the particular analysed patterns. Conclusions: Our preliminary results reveal the existence of retinal markers expression in the ES–D3 embryoid bodies development with a spontaneous differentiation. These observations suggest that ES–D3 cells can serve as a model for understanding the mechanism that regulates the specification of retinal neurons and as a potential source for retinal progenitors.
Keywords: retinal development • retinal degenerations: cell biology