Superoxide modifies AMPA receptors and voltage-gated K+ channels of mouse hippocampal neurons.
Lucifer Yellow CH (LY), a fluorescent membrane-impermeable cell-marker dye, has been routinely loaded into cells through recording electrodes to visualize these cells after electrophysiological investigation. Recently we showed that LY produced superoxide when LY was exposed to light at ordinary intensities for microscopy, and that the resultant superoxide retarded the inactivation of voltage-gated Na+ channels even in the dark. Here, we show that superoxide produced by exposure to light prolongs the duration of action potentials, and increases the magnitude of outward rectifier K+ currents and inward rectifier K+ currents in cultured mouse hippocampal neurons. Superoxide also increases the current response of AMPA receptors, but has no effect on that of kainate and NMDA receptors, GABA(A) receptors, high-voltage activated Ca2+ channels of the hippocampal neurons, nor on 5HT3 receptors of N1E-115 cells. These superoxide effects are irreversible. The addition of superoxide dismutase, an enzyme that selectively decomposes superoxide, to LY-loaded recording electrodes reverses the superoxide effects, but addition of heat-inactivated superoxide dismutase fails to reverse the effects. The application of dithiothreitol, a free radical scavenger, to a bathing solution also reverses the superoxide effects. This shows that superoxide rather selectively modifies ion channels. The effects of exogenous and endogenous superoxide on the superoxide-susceptible channels are discussed.
Takeuchi
Keita
K
Graduate school of Life Science and Systems Engineering, Kyushu Institute of Technology, Hibikino 2-4, Kitakyushu-shi 808-0196, Japan.
Yoshii
Kiyonori
K