Abstract
Retinal degenerative diseases, due to the lack of regeneration systems and self-renewable cells, often lead to visual impairment. Pax6 is a pleiotropic transcription factor and its expression level determines self-renewal status or differentiation of retinal cells. Here, we investigated the fate of simultaneous induction of retinal ganglion cell death and Pax6 overexpression in retro-differentiation of retinal cells and their commitment to re-enter into the cell cycle. Induction of acute retinal ganglion cell death and generation of mouse experimental model was performed by N-methyl D-aspartic acid (NMDA) injection. Recombinant AAV2 virus harboring PAX6 cDNA and reporter gene was injected into untreated and model mouse eyes. Histological analyses, including IHC and retinal flatmounts immunostaining were performed. The number of Ki67+ cells was clearly increased in model mice, presumably due to NMDA treatment and regardless of Pax6 over-expression. Unlike previous studies, Ki67+ cells were found in GCL layer and interestingly ONL cells expressed Sox2 stemness marker after NMDA cytotoxicity. The potential of retinal cells for robust Ki67 expression, after injury, and expression of Sox2, confirmed their intrinsic plasticity and made a vivid prospect for retinal regenerative medicine.
Highlights
In recent years, successful reprogramming of adult cells by ancestral transcription factors was a milestone in cellular biology[10,11,12,13,14]
It has been shown that after retinal injuries in mice, some Pax[6] positive cells migrate from inner nuclear layer (INL) to outer nuclear layer (ONL); this migration does not lead to proliferation and differentiation dislike some lower vertebrates[18]
N-methyl D-aspartic acid (NMDA)-induced RGC damage is a reliable method to generate experimental model. Generation of this model characterizes by rapid RGCs loss followed by gradual reduction in ganglion cell complex thickness (GCC: retinal nerve fiber layer, ganglion cell layer, and inner plexiform layer)
Summary
Successful reprogramming of adult cells by ancestral transcription factors was a milestone in cellular biology[10,11,12,13,14]. The main goal is perturbing gene regulatory network (GRN) to achieve intended phenotype. In these strategies transcription factors are from the effective and convenient choices. Morphogenesis of the eye and is crucial for the development of the central nervous system (CNS), nose, pancreas, and pituitary gland[15,16]. This factor is one of the indicators of RPCs. Its expression level is a determinative factor in self-renewal status or differentiation of retinal cells[17]. We investigated perturbing GRN in inner retina layers with PAX6 overexpression, mediated by adeno-associated virus serotype-2 (AAV-2), and induction of retinal ganglion cell death to analyze expression of cell cycle marker in the mouse retina
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