TTX-sensitive (TTX-S) and TTX-resistant (TTX-R) Na+ currents are expressed in high densities (2–8 channels/microns2) in astrocytes cultured from neonatal rat spinal cord. The two Na+ current types differ up to 1000-fold in their TTX sensitivity and additionally have different steady-state activation (g-V) and inactivation (h infinity) curves. Expression of TTX-S and TTX-R Na+ currents is confined to morphologically distinguishable subtypes of astrocytes, allowing characterization of the two types of Na+ currents in isolation: stellate cells express TTX-S Na+ currents and flat pancake cells express TTX-R Na+ currents. Activation of protein kinase C (PKC) by phorbol 12-myristate 13-acetate (PMA) exhibited different effects on TTX-S and TTX-R Na+ currents. PMA reduced peak TTX-S Na+ currents by 25– 60%; in contrast, PMA potentiated peak TTX-R Na+ currents by 60–150%. These effects developed within minutes, and were typically not reversible. PMA effects were voltage dependent, and shifted steady- state Na+ current activation of TTX-R and TTX-S currents by 6 and 18 mV, respectively, but without affecting their steady-state current inactivation (h infinity). PMA treatment also changed Na+ current kinetics. TTX-R current activation (tau m) was faster and current inactivation (tau h) changed from a single- to a bi-exponential after PMA exposure, suggesting that PKC phosphorylation may have activated formerly quiescent Na+ channels. In contrast, TTX-S current activation (tau m) was unchanged, and current inactivation (tau h), on average, decreased by 50% following PMA exposure.(ABSTRACT TRUNCATED AT 250 WORDS)

1. Astrocytes cultured from rat spinal cord express voltage-activated Na+ channels in high densities (up to 8 channels per microns2). Stellate astrocytes express Na+ currents at all times in vitro. In pancake astrocytes, Na+ channel expression shows a distinct temporal pattern, an absence of channel expression at 1-3 days in vitro (DIV), and peak Na+ channel density at 7-8 DIV. 2. Coculture of spinal cord astrocytes with dorsal root ganglion (DRG) neurons substantially reduces the expression of voltage-activated Na+ channels in both spinal cord astrocyte types. In pancake spinal cord astrocytes, both the percentage of cells expressing Na+ channels and the channel density in Na+ channel-expressing cells are markedly reduced. In stellate spinal cord astrocytes, the percentage of Na+ channel-expressing cells is unchanged, but the Na+ channel density per cell is markedly reduced in coculture. 3. Culturing spinal cord astrocytes in neuron-conditioned media reduces Na+ channel expression in both spinal cord astrocyte types to levels intermediate between coculture and control, suggesting that, at least in part, neuronal effects on Na+ channel expression are mediated by a soluble factor secreted into the media by neurons. 4. As with the expression of voltage-activated Na+ channels, the expression of voltage-activated K+ channels is reduced in both spinal cord astrocyte types cocultured with DRG neurons. The effect is not mimicked by culturing cells in neuron-conditioned media, suggesting that effects on K+ channel expression are mediated by a less stable and more readily degradable factor. 5. Coculture with DRG neurons or culture in neuron-conditioned media do not alter the biophysical properties of voltage-activated Na+ currents in pancake spinal cord astrocytes. Thus steady-state activation, steady-state inactivation, and the time constants of activation and inactivation are virtually unchanged under the various culture conditions.

Compound action potential (CAP) conduction and Na+ channel content were studied in optic nerves from control and myelin-deficient (md) rats. Action potential propagation was approximately five times slower in the md rat, but the action potentials propagated securely and had frequency-following and refractory properties equivalent to control myelinated axons. Tritium-labelled saxitoxin ([3H]-STX) binding in md optic nerve was approximately 30% greater, per wet mass of tissue, than in the control optic nerve. However, calculations of channel density per axon based on previously published anatomical data from md and control optic nerves (Dentinger et al. 1985) show an equivalent number of sodium channels per axon, with an average density of 10 channels micron-2 in md and 11 channels micron-2 in control optic nerve axons. The amplitude of the CAP in both control and md optic nerves was significantly attenuated by 50 nM TTX, precluding the possibility that TTX-insensitive channels are responsible for the action potential in myelinated or amyelinated axons. In addition, the amplitudes of voltage-activated Na+ currents in type I and type II astrocytes cultured from control and md optic nerves were similar, suggesting that the glial component of Na+ channels is not abnormal in the optic nerve of the md rat. These results suggest that myelination (or its absence) may not directly regulate the number of axonal Na+ channels.