STUDIES ON THE MORPHOLOGICAL DIFFERENTIATION OF A MAMMALIAN RETINAL GANGLION CELL LINE

NIETO PAULA S; GUIDO MARIO E

Buenos Aires- Argentina

Congreso; International Congress on Eye Research (ICER); 2006

International Society for Eye Research (ISER)

Recently, Dr. Agarwal’s group establishes a clonal retinal cell line from virally transformed dispersed postnatal rat retinal cells, named RGC-5 which has certain characteristics of retinal ganglion cells (RGCs) based on expression of specific markers such as Thy-1, Brn-3c; Neuritin and NMDA receptor, and on sensitivity to glutamate excitotoxicity and neurotrophin withdrawal (Krishnamoorthy et al., 2001). Thus, these RGC-5 cells appear to be very useful to study various aspects of RGC biology and physiology. However, it still remains uncertain some aspects of these cells regarding their mitotic state and the non-neuronal morphology of them. The principal aim of this work was to investigate the capacity of RGC-5 to differentiate by leaving cell division and inducing a neuronal morphology with the typical neurite formation. To do this, we investigated the response of this cell line to different effectors such as staurosporine (SS), BDNF, NGF and retinoic acid. We found that SS, a protein kinase inhibitor; induce cell differentiation at a lower range of concentration than those previously reported by Frassetto et al. (2006). Concentrations ranging from 10 to 25 ng/mL during 30-60 minutes are sufficient to induce cell arrest and neurite outgrowth and cells survived for more than 24 hours post SS administration. Furthermore, pulses of SS during different times following addition of 10% bovine fetal serum (BFS) to the culture medium showed that cells dedifferentiate and dye by apoptosis in a dose dependent manner according to the nuclear fluorescent staining with DAPI. In addition, treatment with BDNF (80 ng/mL) or retinoic acid (20 ug/mL) produced a slight effect in cell morphology of RGC-5 cells whereas NGF (100 ng/mL) did not show a significant effect.