Effects of in vitro hypoxia and lowered pH on potassium fluxes and energy metabolism in rat brain synaptosomes.
Synaptosomes isolated on isosmotic Ficoll density gradients are an effective model for some aspects of neuronal function. They maintain metabolic energy levels ([ATP]/[ADP] [Pi]) and transplasma membrane electrical potentials very similar to those of neurons in the intact brain. The concentration of K+ in the external medium (K+-sensitive electrode), O2 uptake, and cytochrome c reduction (550 nm minus 540 nm) were simultaneously monitored in synaptosomal suspensions. Oxidative metabolism is the primary source of intrasynaptosomal ATP and at pH 7.4 anaerobiosis results in K+ leakage at 4.5 +/- 0.8 nmol/min/mg protein with glucose as substrate and 10.7 +/- 1.9 nmol/min/mg protein with lactate plus pyruvate (10:1) as substrate. Reintroduction of oxygen initiates complete (ouabain-sensitive) reuptake of K+ at initial rates of 35.4 +/- 3.2 nmol/min/mg protein and 18 +/- 1.7 nmol K+/min/mg protein, respectively. The rates of K+ leakage and reuptake fall when the pH is lowered from 7.4 to 6.0 but recover fully if the pH is raised to the original value. The rates of K+ release and uptake decrease when the Na+ concentration in the medium is decreased and increase when the Ca2+ concentration is decreased. The intrasynaptosomal [K+] under aerobic conditions was 77.3 +/- 3 mM and the calculated K+ diffusion potential was -72 mV. Anaerobic incubation of the synaptosomes from up to 20 min and at pH values from 7.4 to 6.0 did not produce irreversible impairment of any of the measured variables. These results suggest that permanent loss of brain function following prolonged hypoxia and ischemia is not due to irreversible damage to the synapses with respect to these parameters but rather to impairment of some other neuronal functions.
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000693:Anaerobiosis; D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000648:ultrastructure; D002118:Calcium / Q000378:metabolism; D004734:Energy Metabolism; D006863:Hydrogen-Ion Concentration; D008297:Male; D010042:Ouabain / Q000494:pharmacology; D010100:Oxygen / Q000378:metabolism*; D010424:Pentobarbital; D011188:Potassium / Q000378:metabolism*; D051381:Rats; D012964:Sodium / Q000378:metabolism; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism*; D014701:Veratridine / Q000494:pharmacology
D010042:Ouabain; D000244:Adenosine Diphosphate; D014701:Veratridine; D000255:Adenosine Triphosphate; D012964:Sodium; D010424:Pentobarbital; D011188:Potassium; D010100:Oxygen; D002118:Calcium