Different cells subpopulations in the RGC-5 cell line: morphology and calcium responses to ATP

NIETO, P.S; ACOSTA RODRÍGUEZ V. A.; GUIDO MARIO E

Buzios, Rio de Janeiro-Brasil

Congreso; I congreso IBRO/LARC de Neurociencia para Latinoamérica, Caribe y Península Ibérica NeuroLatam; 2008

Sociedad Argentina de Invesigación en Neurosciencias, Sociedade Brasileira de Neurociências e Comportamento, Sociedad Chilena de Neurociencia,Sociedad de Neurociencia del Uruguay

RGC-5 cells are a new cell line derived from postnatal rat retinal ganglion cells (Krishnamoorthy R. et al., 2001, Mol. Brain Res). Although the RGC-5 cells are derived from a single clone; we investigate whether these cells constitute a morphologically homogeneous population. A least tree morphologically different populations of non differentiated RGC-5 in culture were identified by flow cytometry and immunocitochemistry. RGC-5 cells show differential calcium responses to ATP stimulation, some of them, exhibit a high amplitude positive response; other cells display a low amplitude positive response, while a third population shows a null response. These differences could be a direct consequence of heterogeneous expression of different ATP receptors in these cells. Surprisingly, RGC-5 cells do not respond to glutamate stimulation (100 and 200 uM). Staurosporine administration (25ng/mL) induces morphological differentiation that correlates with a change in the flow cytometry pattern at 24 h post staurosporine administration. Interestingly, after 48 h of staurosporine treatment, cells died by apoptosis. The results strongly support the idea that this cell line is not a homogeneous population and this observation may result an important feature to be considered for future studies and for analyzing biochemical responses in these cells.