title: Hypothermia and rewarming injury in hippocampal neurons involve intracellular Ca2+ and glutamate excitotoxicity.
abstract: This study examines the causes of hypothermia and rewarming injury in CA1, CA3, and dentate neurons in rat hippocampal slice cultures. Neuronal death, assessed with propidium iodide or Sytox fluorescence, Fluoro-Jade labeling, and Cresyl Violet staining, depended on the severity and duration of hypothermia. More than 6 h at temperatures less than 12 °C followed by rewarming to 37 °C (profound hypothermia and rewarming, PH/RW) caused swelling and death in large number of neurons in CA1, CA3, and dentate. During PH, [ATP] decreased and [Ca(2+)](I) and extracellular [glutamate] increased, with neuron rupture and nuclear condensation following RW. The data support the hypothesis that neuronal death from PH/RW is excitotoxic, due to ATP loss, glutamate receptor activation and Ca(2+) influx. We found that antagonism of N-methyl-D-aspartate (NMDA) receptors, but not 2-amino-3-(5-methyl-3-oxo-1,2- oxazol-4-yl) propanoic acid or metabotropic glutamate receptors, decreased neuron death and prevented increases in [Ca(2+)](I) caused by PH/RW. Chelating extracellular Ca(2+) decreased PH/RW injury, but inhibiting L- and T-type voltage-gated Ca(2+) channels, K+ channels, Ca(2+) release from the endoplasmic reticulum, and reverse Na(+)/Ca(2+) exchange did not affect the Ca(2+) changes or cell death. We conclude that the mechanism of PH/RW neuronal injury in hippocampal slices primarily involves intracellular Ca(2+) accumulation mediated by NMDA receptors that activates necrotic, but not apoptotic processes.
authors.affiliation: Department of Anesthesia and Perioperative Care, Parnassus Avenue, University of California, San Francisco, CA 94143-0542, USA.
authors.forename: D E
authors.initials: DE
authors.lastname: Warren
authors.forename: P E
authors.initials: PE
authors.lastname: Bickler
authors.forename: J P
authors.initials: JP
authors.lastname: Clark
authors.forename: M
authors.initials: M
authors.lastname: Gregersen
authors.forename: H
authors.initials: H
authors.lastname: Brosnan
authors.forename: W
authors.initials: W
authors.lastname: McKleroy
authors.forename: P
authors.initials: P
authors.lastname: Gabatto
mesh_terms: D000818:Animals; D001831:Body Temperature / Q000502:physiology; D020013:Calcium Signaling / Q000502:physiology*; D018698:Glutamic Acid / Q000502:physiology*; D006624:Hippocampus / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology*; D007036:Hypothermia, Induced / Q000009:adverse effects*; D007424:Intracellular Fluid / Q000378:metabolism / Q000502:physiology*; D009410:Nerve Degeneration / Q000209:etiology / Q000378:metabolism / Q000503:physiopathology*; D009474:Neurons / Q000473:pathology / Q000502:physiology; D009498:Neurotoxins / Q000494:pharmacology; D009924:Organ Culture Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D009498:Neurotoxins; D018698:Glutamic Acid