Regulation of fos and jun Immediate-Early Genes by Redox or Metabolic Stress in Cardiac Myocytes

We have previously demonstrated coordinate inductions of c-fos, c-jun, jun B, and jun D in cardiac myocytes exposed to hypoxia for 2 to 4 hours. Induction of these transcripts occurred before any significant loss of intracellular ATP. In the present study, the origin of the signal(s) that regulates immediate-early gene induction was investigated by comparing the effects of hypoxia with those of the metabolic inhibitors cyanide, deoxyglucose and cyanide combined, and iodoacetic acid. Cyanide, an inhibitor of oxidative metabolism, closely mimicked the metabolic effects of hypoxia, with elimination of oxygen consumption, increased lactate production, and minimal decline in ATP levels under both conditions. Compared with hypoxia, cyanide mediated small transient inductions of fos and jun transcripts that followed a different time course. The combination of cyanide and deoxyglucose resulted in inhibition of lactate production as well as respiration, and ATP dropped rapidly to 20% of control levels. The loss of intracellular ATP was followed by fourfold inductions of c-fos and c-jun with minor changes in jun B and jun D transcript levels. Similarly, iodoacetic acid caused a major (90%) loss of ATP and irreversible cell damage as measured by leakage of creatine phosphokinase enzyme and loss of membrane arachidonic acid; ATP loss was followed by fivefold to sevenfold inductions of c-fos, c-jun, and jun B transcripts. We conclude that the hypoxic stress response in neonatal myocytes, which occurs before ATP depletion, cannot be fully accounted for by inhibition of oxidative metabolism or by factors related to metabolic switching. In contrast, the stress response associated with blockage of both aerobic and anaerobic energy metabolism coincides with and may be related to the loss of cellular ATP and structural cell damage.