INTRACELLULAR CA2+ CONCENTRATION AND PHI DURING METABOLIC INHIBITION

To study the changes in intracellular Ca2+ concentration ([Ca2+]i) and pH (pH(i)) during metabolic inhibition, rat ventricular myocytes were dual loaded with the acetoxymethyl esters of fura-2 (fura-2/AM) and 2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF/AM) and perfused with 2 mM sodium cyanide (NaCN). The percent of rod-shaped cells was 30% of the control after 30 min NaCN in the absence of glucose. [Ca2+]i increased from 82 +/- 8 to 151 +/- 25 (SE) nM (P < 0.05) when cells were shortened, and to 421 +/- 106 nM (P < 0.05) when cells were rounded. There was a positive relationship between pCa(i) and pH(i) (r = 0.425, P < 0.01). When 50 mM glucose was added during NaCN, there were no significant changes in [Ca2+]i and the percent of rod-shaped cells after 30 min. The pH(i) of rod-shaped cells, however, decreased to 6.95 +/- 0.03 (P < 0.01). Although the acidic solution (70% O2-30% CO2) decreased pH(i) to 6.90 +/- 0.05 (P < 0.01), there were no changes in the cell shape or [Ca2+]i. The addition of NaCN for 30 min decreased the rod-shaped cells to 18% of the control. Mild acidosis did not cause the changes in cell shape or [Ca2+]i. There was also no protection of the NaCN-induced changes in cell shape by intracellular acidosis. It is likely that the changes in cell shape during metabolic inhibition were related to the depletion of metabolic energy and the increase in [Ca2+]i.