Characterization of sodium currents in mammalian sensory neurons cultured in serum-free defined medium with and without nerve growth factor

Abstract

The influence of nerve growth factor (NGF) on Na currents of rat dorsal root ganglia (DRG) was studied in neurons obtained from newborns and cultured for 2-30 hr in serum-free defined medium (SFM). Cell survival for the period studied was 78-87% both with and without NGF. Na currents were detected in all cells cultured for 6-9 hr. They were also detected after 2 hr in culture in 21.5% of the cells cultured without NGF (-NGF cells), and in 91.5% of the cells cultured with NGF (+NGF cells). Current density of the -NGF cells was 2.3 and 2 pA/microns 2 after growth for 2 and 6-9 hr, respectively, compared to 3.0 and 3.9 pA/microns 2 for the +NGF cells. The +NGF cells were separated into fast (F), Intermediate (I) and slow (S) cells, based on the Na current they expressed, while -NGF cells were all of the I type. F, I and S currents differed in their voltage-dependent inactivation (Vh50 = -79, -28 and -20 mV), kinetics of inactivation (tau h = 0.55, 1.3 and 7.75 msec), and TTX sensitivity (Ki = 60, 550 and 1100 nM). All currents were depressed by [Ca]0 with a KdCa of 22, 17 and 8 mM for F, I and S currents, respectively. Current density of F and S currents was 5.5 and 5 pA/micron 2 for the I current. The concentration-dependent curve of I current vs. TTX indicated that I current has two sites: one with F-like and another with S-like Ki for TTX. Hybridization of F and S currents yielded I-like currents. Thus, the major effect of NGF on Na currents in SFM is the acceleration of Na current acquisition and diversity, reflected in an increase of either the S or F type in a cell.