Importance of glycolytically derived ATP for Na+ loading via Na+/H+ exchange during metabolic inhibition in guinea pig ventricular myocytes.
The increase in the intracellular Na+ concentration ([Na+](i)) during myocardial ischaemia is crucial for ischaemia/reperfusion cell injury, and the cardiac subtype of the Na+/H+ exchanger (NHE-1) has been shown to be a major pathway for Na(+) loading. While the importance of glycolytically derived ATP for the optimal functioning of membrane transporters and channels has been suggested, whether NHE-1 is actually activated during myocardial ischaemia remains controversial. Here we examined whether the activity of NHE-1 is predominantly dependent on intracellular ATP generated by glycolysis, and whether the additional inhibition of glycolysis can affect the increase in ([Na+](i)) during the inhibition of oxidative phosphorylation in intact guinea pig ventricular myocytes. The selective inhibition of glycolysis by 2-deoxyglucose prevented the recovery of intracellular pH and the transient increase in ([Na+](i)) following intracellular acidosis induced by a NH(4)Cl pre-pulse. During severe metabolic inhibition (SMI; induced by amobarbital and carbonyl cyanide m-chlorophenylhydrazone in a glucose-free perfusate), most myocytes changed from rod-shaped to contracted forms by approximately 15 min. ([Na+](i)) increased linearly until rigor contracture occurred, but after rigor contracture the rate of increase was blunted. The increase in ([Na+](i)) during SMI was suppressed significantly by an inhibitor of NHE-1, hexamethylene amiloride. The increase in the intracellular Mg(2+) concentration, which can reciprocally indicate depletion of intracellular ATP, was small during the initial 10 min of SMI, but became larger from just a few minutes before rigor contracture. In the presence of 2-deoxyglucose, the time to rigor during SMI was shortened, but the increase in ([Na+](i)) before rigor contracture was not significant, and was much less than that in the absence of 2-deoxyglucose. It is concluded that ATP generated by glycolysis is essential to activate NHE-1, and that the dependence of NHE-1 on glycolysis might affect the increase in ([Na+](i)) observed during myocardial ischaemia.
Satoh
H
H
Division of Cardiology, Internal Medicine III, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu 431-3192, Japan. satoh36@hama-med.ac.jp
Sugiyama
S
S
Nomura
N
N
Terada
H
H
Hayashi
H
H
D000255:Adenosine Triphosphate / Q000502:physiology*; D000818:Animals; D018929:Cell Culture Techniques; D005260:Female; D006019:Glycolysis; D006168:Guinea Pigs; D006863:Hydrogen-Ion Concentration; D017202:Myocardial Ischemia / Q000378:metabolism* / Q000473:pathology; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D012964:Sodium / Q000378:metabolism*; D017923:Sodium-Hydrogen Exchangers / Q000502:physiology*
D017923:Sodium-Hydrogen Exchangers; D000255:Adenosine Triphosphate; D012964:Sodium