Intracellular calcium during photodynamic permeabilization of cardiomyocytes

The purpose of this study was to determine whether an early increase in [Ca2+]i preceding generalized lysis of cardiomyocytes occurred during photodynamic permeabilization. A method was developed which facilitated the simultaneous measurement, in real time, of permeabilization of the sarcolemma to Ca2+ and Mn2+ during photodynamic action. Quin-2 loaded cells were illuminated in the presence of erythrosin B and the change in the fluorescence emission of the calcium-quin-2 complex was used to measure the rate and extent of change in [Ca2+]i. The same system was used in the presence of extracellular Mn2+ to determine how quickly the cardiomyocytes became permeable to either Mn2+ or quin-2. Calcium ions were observed to enter the myocytes prior to permeabilization of the sarcolemma to either Mn2+ or quin-2, and thus before membrane lysis. Lysis of cardiomyocytes did not appear to be dependent upon increases in [Ca2+]i. Controls were performed to rule out fluorescent artifacts. Reperfusion injury and photodynamic therapy involve both the production of free radicals and an early increase in [Ca2+]i. This study demonstrates a direct correlation between the production of reactive oxygen species and prelytic increases in [Ca2+]i in neonatal cardiomyocytes and demonstrates that this phenomenon may be common to many cell types.