Neural excitability, synaptic transmission and neuron-glia interactions
Author: Maximiliano R. Hurtado Paso | Email: maxihurtado777@gmail.com
Maximiliano R. Hurtado Paso1°, Verónica Bisagno2°, Francisco J. Urbano,
1° IFIBYNE-CONICET, University of Buenos Aires, Argentina
2° School of Biomedical Sciences, Austral University, Buenos Aires, Argentina
Our group has previously used N2a mouse neuroblastoma cell line to study voltage-gated channel expression, signaling pathways, neurite growth and histone deacetylases mediated (HDACs) neuronal differentiation. N2a cells are neural precursors that can change their morphology under a low fetal bovine serum cell culture condition; extending their cytoplasm forming neurites in bipolar or multipolar shapes. Understanding the mechanisms underlying N2a cells ability to differentiate into neurons is important to develop treatments to prevent human neuroblastoma spreading. N2a cells were cultured for 4 days in vitro (d.i.v.) in a DMEM-0.5% FBS condition and treated with a cholinergic agonist carbachol (CAR, 50 uM) or dbcAMP (50 uM). Some experiments combined CAR/dbcAMP with a low concentration (50 uM) of MS275/MC1568 class I/IIa HDACs inhibitors respectively, or DMSO. Whole cell patch-clamp recordings and neurite length/type characterization among treatments were performed to study N2a differentiated neuron-like characteristics. Our results showed that N2a cells were able to adopt a neuron-like differentiation phenotype after 4 d.i.v. of DMSO or CAR/dbcAMP. Density of voltage-gated sodium and potassium currents was enhanced after CAR treatment (One way ANOVA). These results suggest that N2a cells can be differentiated in the presence of cholinergic agonists adopting a similar neuron-like morphology previously described for dbcAMP treatments.