Resting Ca2+ influx does not contribute to anoxia-induced cell death in adult rat cardiac myocytes.
Calcium has been proposed as a primary influence on cell death during ischemic episodes in myocardial cells. One component of calcium entry into a cell is resting calcium influx. This basal movement of calcium is blocked by 100 micromol/L gadolinium chloride (GdCl3) in cardiac myocytes. Therefore, GdCl3 should be cardioprotective under anoxic conditions. To test this, cardiac myocytes isolated from adult male rats were subjected to anoxia (100% N2) in the presence or absence of 100 micromol/L GdCl3 in one of 2 ways. In the first method, cells were suspended in media and rendered anoxic for 0, 30, and 60 min, after which cell morphology and viability were scored. After 60 min of anoxia, rod-shaped cells accounted for 46% +/- 4% of total cells (viability 81%); 10 min of reoxygenation markedly reduced rod-shaped cells to 27% (viability 72%). GdCl3 in the medium did not protect the cells (anoxic rods 49%, reoxygenated rods 30%, viability 77%). In the second method, cells were attached to a laminin substrate, rendered anoxic, and then videotaped for up to 6 h. In this system, cells maintained their shape for some time after the onset of anoxia, and then began to 'die' (i.e., to take on either a rigor form or hypercontracted form) at a measurable rate. Time to onset of 'death' (t0), time to 50% and 100% 'death' (t50 and t100), and rate of 'death' were used to measure anoxic damage. Without GdCl3, cells on average began to die 115 +/- 32 min after the onset of anoxia (t0); they died at an average rate of 0.046 cells/min. t50 was achieved in 149 +/- 42 min, t100 in 183 +/- 54 min. Addition of 100 micromol/L GdCl3 did not affect any of these parameters. We concluded that GdCl3 was not cardioprotective for anoxic myocytes and that cell damage generated by anoxia could not be attributed to resting calcium influx.
Mont
Meghan R
MR
Department of Physiology, A.T. Still University of Health Sciences, 800 West Jefferson Street, Kirksville College of Osteopathic Medicine, Kirksville, MO 63501, USA.
Carlson
C George
CG
Geisbuhler
Timothy P
TP
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020746:Calcium Channels, L-Type / Q000502:physiology; D016923:Cell Death; D015687:Cell Hypoxia; D005682:Gadolinium / Q000494:pharmacology; D008297:Male; D008345:Manganese / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism* / Q000473:pathology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013779:Tetrodotoxin / Q000494:pharmacology
D020746:Calcium Channels, L-Type; D008345:Manganese; D013779:Tetrodotoxin; D005682:Gadolinium; D002118:Calcium