Journal of Neurophysiology, Vol 69, Issue 3 819-831, Copyright © 1993 by APS

ARTICLES

Ion channels in spinal cord astrocytes in vitro. III. Modulation of channel expression by coculture with neurons and neuron-conditioned medium

C. L. Thio, S. G. Waxman and H. Sontheimer
Department of Neurology, Yale University School of Medicine, New Haven 06510.

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.

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