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Lei Gu, Joseph Caprioli, Natik Piri; Temporal and spatial expression patterns of Rbfox1 and Rbfox2 during mouse retinal development. Invest. Ophthalmol. Vis. Sci. 2018;59(9):5494.
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© ARVO (1962-2015); The Authors (2016-present)
Rbfox proteins regulate alternative splicing, mRNA stability and translation. Here, we have characterized expression of Rbfox1 and Rbfox2 in adult and developing retinas and identified Rbfox1-regulated genes that may be important for normal retinal function.
E12, E15, P0, P5, P10, P12, P14, P15 and P21 retinal sections were analyzed by immunohistochemistry with antibodies specific for Rbfox1 and Rbfox2. Retinal flatmount double immunofluorescence was used to quantify Rbfox1 and Rbfox2 positive cells. The retinal transcriptome was analyzed with RNA sequencing.
In adult retinas, Rbfox1 and Rbfox2 were expressed in all types of retinal ganglion cells (RGCs) as well as in subsets of amacrine cells (ACs) within the inner nuclear and ganglion cell layers. Rbfox2 expression was also observed in horizontal cells (HCs). In developing retinas at E12 and E15, Rbfox1 was localized to the cytoplasm of differentiating RGCs and ACs. Rbfox1 expression shifted from cytoplasmic to predominantly nuclear starting at around P0; nuclear localization was more prominent at P5. By P10, Rbfox1 distribution in RGCs and ACs was predominantly nuclear and remained as such in fully differentiated retina. The shift in Rbfox1 subcellular distribution can be correlated with the stage II spontaneous retinal waves of excitation, which in mice takes place during first 1–2 postnatal weeks. Neuronal depolarization has been shown to regulate Rbfox1 alternative splicing, resulting in the increased expression of the splicing-active nuclear isoform. Rbfox1 strong dendritic expression was observed between P0 and P5 in the IPL; by P10 it was reduced to the background level. Rbfox2 expression in RGCs, ACs and HCs showed no dynamic subcellular changes and remained nuclear throughout retinal development. Transcriptome analysis of Rbfox1 knockout (KO) retinas identified several genes, including Vamp1, Vamp2, Snap25, Trak2, and Slc1A7 that are involved in establishing neuronal circuits and synaptic transmission.
Comprehensive analysis of Rbfox1 and Rbfox2 expression in adult and developing retinas together with RNAseq data and our earlier observation on depth perception deficiency in Rbfox1 KOs suggest important roles of these proteins in regulation of splicing and expression of genes involved in the differentiation and normal function of RGCs and ACs.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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