The presynaptic modulation of glutamate release and the membrane dysfunction induced by in vitro ischemia in rat hippocampal CA1 neurons

Abstract

Superfusion with an oxygen and glucose deprived medium (in vitro ischemia) of rat hippocampal CA1 pyramidal neurons in tissue slices produced a rapid depolarization within 5 min and thereafter showed no functional recovery (irreversible membrane dysfunction), even if oxygen and glucose were reintroduced. We previously suggested that such a rapid depolarization is triggered by the accumulation of extracellular glutamate (Glu). As a result, we examined the effects of either the activation or inhibition of presynaptic receptors, which modulate Glu release from the nerve terminal, on the potential change produced by in vitro ischemia. The adenosine A 1 receptor antagonist, 8-cyclopenthyl theophylline, A 2a receptor antagonist, ZM241385, and A 2b receptor antagonist, alloxazine, did not significantly alter either the latency or the maximal slope of the rapid depolarization. In addition, the GABA B receptor antagonist, 2-hydroxysaclofen, or the metabotropic Glu receptor type 4 antagonist, alpha-methylserine-O-phosphate, did not change either the latency or the maximal slope. The adenosine A 1 receptor agonist, 2-chloro-N 6-cyclopentyladenosine, A 2a receptor agonist, CGS2168, or A 2b receptor agonist, 5′-(N-ethylcarboxamido)-adenosine, did not affect these parameters either. None of these drugs restored the membrane potential to the pre-exposure level after the reintroduction of oxygen and glucose. Simultaneous intracellular recordings from CA1 and CA3 pyramidal neurons in the same slices revealed the membrane of the CA3 neurons to be hyperpolarized when a rapid depolarization occurred in the CA1 neurons. These results suggest that presynaptic Glu release does not accelerate during the generation of the rapid depolarization induced by in vitro ischemia.