Induced Hippocampal Neuron Protection in an Optimized Gerbil Ischemia Model: Insult Thresholds for Tolerance Induction and Altered Gene Expression Defined by Ischemic Depolarization

Abstract

Preconditioning of hippocampal CA1 neurons was evaluated in a gerbil model of transient global ischemia using extracellular recording of DC potential shifts characteristic of ischemic depolarization to precisely define the duration of both priming and test insults. Brief ischemia resulting in depolarizations of 2.5 to 3.5 minutes consistently induced maximal tolerance (95% protection) against subsequent challenges 2 days later with an approximate doubling of the insult duration required for complete CA1 neuron loss from 6 to 12 minutes depolarization when evaluated 1 week after the test insult. Significant protection persisted at 2 months survival, although the apparent injury threshold regressed to approximately 8 minutes, indicating delayed progression of injury after longer test insults. In situ hybridization was used to evaluate depolarization thresholds for induction of mRNAs encoding the 70 kDa heat shock/stress protein, hsp72, as well as several immediate-early genes (c-fos, c-jun, junB, and junD). Immediate-early genes were prominently expressed after short insults inducing tolerance, whereas appreciable hsp72 induction only occurred after insults approaching the threshold for neuron injury. These results establish an ischemic preconditioning model with the predictability needed for mechanistic studies and demonstrate that prior transcriptional activation of the postischemic heat shock response is not required for expression of delayed tolerance.