Sodium dependence of the nerve growth factor--regulated hexose uptake in chick embryo ganglionic cells

Embryonic dorsal root ganglionic cells, when incubated in vitro in the absence of nerve growth factor (NGF) undergo a general metabolic degeneration which is preceded by certain changes in permeation properties. Previous studies demonstrated that NGF can rapidly modulate permeation properties which regulate the availability to the cell of an important energy source, glucose. Hexose uptake was determined by measuring the ability of the cells to accumulate [3H]labeled 2-deoxy-D-glucose. The work reported here shows that the NGF-dependent portion (about one-third) of the total specific hexose uptake was also dependent on the presence of Na+, with the apparent uptake constant (Kt) for deoxyglucose varying inversely with an external Na+ concentration of 70-140 mM; Vmax was unaffected in this range. Preincubation of ganglionic cells with 10 mM ouabain for 15-60 min, followed by a pulse with [3H]-deoxyglucose, also resulted in 50-95% reduction of the NGF-sensitive uptake. A similar pretreatment of cells with veratridine gave a 25-50% reduction in uptake. The NGF-controlled hexose uptake was also energy dependent, being diminished 50-95% after a 30-90 min preincubation with 2 mM 2,4-dinitrophenol. Uptake activities for other substrates (alpha-aminoisobutyric acid, uridine) which exhibited NGF regulation were likewise Na+-sensitive. These results indicate that availability of major energy substrates to NGF-dependent dorsal root ganglionic neurons is controlled by sodium gradients across their membranes. It is conceivable that NGF provides for maintenance and development of its target neurons by acting on such sodium gradients and, consequently, regulating the intake of essential nutrients.