Recovery of cultured rat neonatal myocytes from hypercontracture after chemical hypoxia.
Cardiac myocytes exposed to anoxia hypercontract into a blebbed, rounded mass. Such hypercontraction is usually considered a manifestation of irreversible injury. Here, we studied functional recovery, long-term viability and ATP levels of cultured neonatal rat cardiac myocytes after metabolic inhibition with cyanide and 2-deoxyglucose, a model of 'chemical hypoxia' which mimics the ATP depletion and reductive stress of hypoxia. After addition of inhibitors, 5-day cultured myocytes ceased spontaneous contractions within 1-2 min, blebbed and hypercontracted after 35 min, and lost viability after 100 min as assessed by nuclear labelling with propidium iodide. 11-day cultured myocytes exhibited a similar progression of injury. When the metabolic inhibitors were removed, spontaneous contractions resumed after an average of 11 min in non-hypercontracted myocytes. In hypercontracted myocytes, weak contractions resumed after 35 min, regardless of the length of time spent in the hypercontracted state, but restoration of strong synchronous contractions took hours. Recovering cells remained viable and exhibited spontaneous contractions through 24 hours of observation, whereas contractility never returned in propidium iodide-labelled cells. ATP levels decreased rapidly after chemical hypoxia and were partially restored upon washout of inhibitors. ATP recovery was similar in non-hypercontracted and hypercontracted cells. Thus, contractile dysfunction during recovery in hypercontracted cells was not due to lack of regeneration of ATP. In conclusion, hypercontracture is not a manifestation of irreversible injury. Contractile dysfunction following recovery from hypercontracture observed here may represent 'in vitro stunning'.
Bond
J M
JM
Department of Cell Biology & Anatomy, University of North Carolina, Chapel Hill 27599.
Herman
B
B
Lemasters
J J
JJ
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn / Q000378:metabolism*; D015687:Cell Hypoxia / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D003847:Deoxyglucose / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D009119:Muscle Contraction / Q000502:physiology; D009132:Muscles / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D012966:Sodium Cyanide / Q000494:pharmacology
D000255:Adenosine Triphosphate; D003847:Deoxyglucose; D012966:Sodium Cyanide