Confocal imaging of protein kinase C isozymes and pHi changesin cardiac cells.

In our experiments on isolated rat cardiac cells we studied the intracellular activation and translocation of protein kinase C (PKC) isozymes and pH changes using confocal microscopy. We detected the delta, kappa and mu isozymes of PKC in cardiac cells stimulated with phorbol ester, alpha-adrenergic agonist phenylephrine and different sigma receptor ligands. The histochemical imaging was performed on a PhioBos 1000 confocal microscope (Sarastro Inc., Ypsilanti, MI) equipped with Zeiss Universal Optics argon laser illumination. The changes of intracellular pH (pHi) following the incubation of the cells with the sigma receptor ligands were studied in the cells preincubated with pH-sensitive fluorescent indicator BCECF and then examined by a confocal scanning fluorescence microscope (ACAS 470 Interactive Laser Cytometer, Meridian). Confocal microscopy was proved superior and irreplaceable in studying the dynamics of intracellular signal transduction.