CEFYBO   02669
CENTRO DE ESTUDIOS FARMACOLOGICOS Y BOTANICOS
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
NMDA induces pluripotent genes in retinal Müller cells in vitro.
Autor/es:
NATALIA BELTARMONE; MAGALÍ MASPERO; CLAUDIA CAPURRO; FRANCISCO SANHUEZA; DAFNE M SILBERMAN
Reunión:
Simposio; 3rd symposium on physiology and pathology of neuroglia; 2020
Resumen:
Some subpopulations of Müller glial cells express markers of pluripotency suggesting that they might serve as a source of retinal neurons following injury. By using an in vitro model of neurotoxic damage, we aimed to identify the subpopulation/s of Müller cells with reprogramming and regenerating capacity.The retinal derived glial cell line MIO-M1 was cultured with NMDA and the regeneration capacity was assayed by determining protein and expression levels of several pluripotent genes (Oct4, Sox2, Sox9, Nanog, Ascl1, Lin28b). All proteins analyzed showed increased levels when treated with NMDA 100 μM. SOX9, OCT3/4 and LIN28b also showed increased levels at NMDA 50 M while NANOG showed decreased levels, both with 50 and 500 μM, when compared with untreated cells. Expression levels of all genes were analyzed by qPCR and showed an increase when exposed to NMDA 100 μM. Results for SOX9 were confirmed by immunofluorescence and need to be validated for the other proteins. According to these results, we chose SOX9 as a pluripotency marker to sort the cells and perform a transcriptomic study by single cell RNA-sequencing analysis. The first attempt made with the subpopulation of sorted cells yielded very poor-quality RNA according to the standards required for this kind of analysis. We are working on the optimization of the procedure to obtain quantifiable samples.We show preliminary data that suggest that Müller cells subjected to neurotoxic damage in vitro can induce the expression of pluripotent genes that in turn would be able to mount a regenerative response.