Tyrphostin AG879 and c-Src inhibitors reduced neurite outgrowth induced by stimulation of Ang II AT2 receptors in SH-SY5Y neuroblastoma cells. XXXIV Reunion Anual Sociedad Argentina de Investigacion en Neurociencias (SAN).

ALVAREZ S.E.; BLANCO H. M; CIUFFO G.M.; BANCHIO C

Carlos Paz-Córdoba

Congreso; XXXIV Reunion Anual Sociedad Argentina de Investigacion en Neurociencias (SAN); 2019

SH-SY5Y is a neuroblastoma cell line used as model of Parkinson disease, Alzheimer and differentiation. The signaling mechanism of neurite outgrowth induced by Ang II AT2 receptors and the interaction with NGF receptors remains unclear. We evaluated neurite outgrowth under differentiation conditions in SH-SY5Y cells, in the presence of different inhibitors: UO126 (MEK inhibitor), LY294002 (PI3K inhibitor) and PP2 (c-Src inhibitor). Only PP2 was able to reduce neurite outgrowth induced by the AT2 receptor´s agonist CGP42112A, supporting the role of c-Src in the signaling pathway, without participation of MAPK or PI3K. Activation of c-Src was confirmed by phosphorylation at residue Y416, a rapid response, followed by Y416 dephosphorylation and Y527 phoshorylation (deactivation of c-Src), suggesting a physiological response. The expression level of Ang II AT2 receptors increased with differentiation, induced by stimulation with CGP42112A, RA or NGF. We evaluated the participation of the TrkA receptor, by using AG879, a specific inhibitor. AG879 clearly inhibited neurite outgrowth, following stimulation with either Ang II or CGP42112A. Taken together, this observation with the effect of CGP42112A in neurite outgrowth induction and increased expression level of AT2 receptors following neurodifferentiation, these results suggest an interaction between both receptors, AT2 and NGF. We further identified the participation of c-Src as a key player in the signaling pathway.