Calcium and increase excitability promote tolerance against anoxia in hippocampal slices.
We have previously demonstrated that anoxic preconditioning (APC) protects against a subsequent otherwise 'lethal' anoxic insult in hippocampal slices. Tested here are two hypotheses: (a) APC requires calcium to improve electrical recovery in hippocampal slices; and (b) mild excitation promotes preconditioning neuroprotection. Control hippocampal slices were given a single 'test' anoxic insult followed by reoxygenation. Experimental slices were preconditioned by three short anoxic insults of 1 min separated by 10 min of reoxygenation. At 30 min after the third 'conditioning' insult, slices underwent a 'test' anoxic insult [1 min of anoxic depolarization (AD)], and then slices were reoxygenated. Evoked potentials (EPs) were recorded throughout the experiment. In other slices, APC was emulated by inducing spreading depression (as determined by a negative DC shift) with KCL or by inducing increased neuronal excitability with the excitatory agent 8-cyclopentyl-1, 3-dipropylxanthine (DPCPX) (an adenosine A1 receptor blocker). 'Test' anoxic insults lasted 2 min of AD in these groups. To determine the role of calcium during APC, extracellular CaCl2 was decreased to 0.5 mM but only during the APC episodes ('test' anoxia, 1 min of AD). EP amplitudes recovered significantly better after anoxia in preconditioned slices, and in KCl- and DPCPX-treated slices (147.2+/-33.3, n=8, **p<0.01, 71.7+/-13.5, n=7, **p<0.01, and 117.8+/-37.3, n=5, ***p<0.001, respectively) compared to controls. Decreases in extracellular CaCl2 during APC blocked the recovery of EPs after 'test' anoxia (80.6+/-23.0, n=8). These data confirm that increases in excitability can emulate APC. These data also demonstrate that calcium influx during preconditioning is required for the induction of tolerance during APC.
Pérez-Pinzón
M A
MA
Department of Neurology D4-5, University of Miami School of Medicine, P.O. Box 016960, Miami, FL 33101, USA. mperez@neuron.med.miami.edu
Born
J G
JG
Centeno
J M
JM
D000222:Adaptation, Physiological / Q000502:physiology*; D000818:Animals; D002118:Calcium / Q000502:physiology*; D003213:Conditioning, Psychological / Q000502:physiology; D013181:Cortical Spreading Depression / Q000502:physiology; D004594:Electrophysiology; D005071:Evoked Potentials / Q000187:drug effects / Q000502:physiology; D006624:Hippocampus / Q000187:drug effects / Q000503:physiopathology*; D000860:Hypoxia / Q000503:physiopathology*; D066298:In Vitro Techniques; D008297:Male; D011189:Potassium Chloride / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D014970:Xanthines / Q000494:pharmacology
D014970:Xanthines; D011189:Potassium Chloride; C051360:1,3-dipropyl-8-cyclopentylxanthine; D002118:Calcium