Enduring changes in Purkinje cell electrophysiology following transient exposure to AMPA: correlates to dark cell...

Purkinje cells (PCs) are selectively vulnerable to alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-mediated delayed toxicity that is manifested as dark cell degeneration (DCD) rather than necrosis. The purpose of the present study was to utilize electrophysiologic changes induced by AMPA to gain mechanistic insights into its cytotoxic actions. The whole-cell configuration of the patch clamp technique was used to record spontaneous electrical activity and ionic currents of Purkinje neurons from cerebellar slices using an experimental paradigm known to produce DCD in response to AMPA. Initial electrophysiologic responses to AMPA consisted of a large transient depolarization and inward current that declined by 75% 20 min into the 30-min exposure to 30 microM AMPA. Cellular responses temporarily continued towards basal levels following removal of AMPA. A sustained membrane depolarization (and underlying persistent inward current), an abundance of apparent excitatory synaptic events, and loss of electro- and chemoresponsiveness were observed 60-75 min into the expression phase (following AMPA removal). These events correspond temporally to the development of DCD in Purkinje cells and may represent an electrophysiological signature of AMPA receptor-mediated delayed neurotoxic events. Antagonists of the AMPA receptor present concomitantly with AMPA are known not to affect DCD and failed to alter the electrophysiologic changes. The secondary depolarization and loss of electroresponsiveness were prevented by antagonists present after removal of AMPA, at a time when DCD also is prevented. Electrical clamping of the PC membrane to equivalent depolarized membrane potentials (V(m)s) obtained with AMPA failed to elicit any long lasting alterations in PC physiology. Collectively, morphological and electrophysiological data indicate that induction of DCD is not strongly dependent on ionotropic mechanisms elicited by AMPA receptors, but that expression of DCD does possess an ionotropic element.