Inhibition of alpha 2/alpha 3 sodium pump isoforms potentiates glutamate neurotoxicity.
Excessive stimulation of neurons by glutamic acid initiates a destructive cascade of ion fluxes, cellular swelling, and death. Homeostatic mechanisms which rectify these disturbances depend largely upon transmembrane ion gradients maintained by Na+,K(+)-ATPase (NaP). We proposed that the neurotoxicity of glutamate is enhanced when the NaP capacity is exceeded, and therefore, that the degree of neuronal death varies inversely with endogenous NaP activity. To test this concept, we directly reduced NaP activity in cultured rat telencephalic cells using either the specific inhibitor ouabain, or dcAMP, and assessed whether these treatments increased glutamate-induced neuronal death. Since rodent NaP catalytic subunits possess both low (alpha 1) and high (alpha 2/alpha 3) affinity for ouabain, we were able to inhibit selectively the alpha 2 (principally glial) and alpha 3 (neuronal) catalytic subunits without affecting the alpha 1 isoform. Brief exposures (5-60 min) to high ouabain concentrations (1-10 mM), which blocks the activity of all three catalytic subunits, killed differentiated neurons but spared glia. In contrast, differential inhibition of the alpha 2/alpha 3 isoforms (by 1 microM ouabain) was not of itself toxic, but produced a supersensitivity to glutamate. [3H]Ouabain binding studies confirmed that the glutamate neurotoxicity observed varied inversely with the degree of NaP inhibition. Further, this relationship was not absolutely dependent upon ouabain, since reductions in alpha 2/alpha 3 pump activity induced by dcAMP also amplified glutamate toxicity. We conclude that inhibition of neuronal NaP with high affinity for ouabain is not lethal to unstimulated cells, but markedly increases susceptibility to glutamate excitotoxicity.(ABSTRACT TRUNCATED AT 250 WORDS)
Neuroendocrine Program, Yale School of Medicine, New Haven, CT 06510.
M L
ML
Brines
R J
RJ
Robbins
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004357:Drug Synergism; D005971:Glutamates / Q000633:toxicity*; D018698:Glutamic Acid; D007527:Isoenzymes / Q000037:antagonists & inhibitors*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009474:Neurons / Q000187:drug effects*; D010042:Ouabain / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012414:Rubidium Radioisotopes; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors* / Q000502:physiology*; D013687:Telencephalon / Q000166:cytology / Q000187:drug effects
D005971:Glutamates; D007527:Isoenzymes; D012414:Rubidium Radioisotopes; D018698:Glutamic Acid; D010042:Ouabain; D007770:L-Lactate Dehydrogenase; D000254:Sodium-Potassium-Exchanging ATPase