Stimulus-dependent labeling of cultured ganglionic cell with [14C]2-deoxyglucose

The authors prepared cultures of dissociated cells from the ciliary (CG) and dorsal root ganglion (DRG) of 10-12-day-old chick embryos, and applied [14C]2-deoxyglucose ([14C]2-DG) to the cultured cells to examine the effects of stimulation on the labeling with [14C]2-DG at the single cell level. Electrical current stimulation increased [14C]2-DG uptake in CG and DRG neurons. The increase depended on frequency of the stimulation. These effects were potentiated by the application of tetraethylammonium, but suppressed by tetrodotoxin. Externally applied potassium ions increased the [14C]2-DG uptake in the CG cell, depending logarithmically on the concentration of applied KCl. The concentration-dependent increase agreed with potassium effect on the equilibrium potential. For CG cells, acetylcholine (ACh), glutamate, γ-aminobutyric acid (GABA) and glycine induced remarkable increases of the [14C]2-DG uptake, while dopamine did not induce any change. For DRG cells, GABA and glycine facilitated the [14C]2-DG uptake, while ACh, glutamate and dopamine did not have any significant effects on it. These facilitatory actions of neurotransmitters on the [14C]2-DG uptake are mostly consistent with the excitatory effects of the substrates on both CG and DRG cells in culture. The results suggest that the [14C]2-DG uptake in single cells is intimately correlated with action potential generation and change in the resting potential. (Auth.)