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In rat ventricular myocytes chemically skinned with saponin, a local rise of [Ca2+] was achieved by two-photon excitation photolysis (TPP) of the caged Ca2+ compound 1-(2-nitro-4,5-dimethoxyphenyl)-N,N,N',N'-tetrakis[(oxycarbonyl)methyl]-1,2-ethanediamine (DM-nitrophen). Confocal Ca2+ images, by use of fluo-3, were simultaneously collected. TPP of DM-nitrophen induced Ca2+ waves propagating over the myocyte, and the local rise of [Ca2+] at the site of photolysis sustained for 50-60 ms. These TPP-induced Ca2+ events were completely suppressed by ryanodine (10 microM), suggesting that Ca2+ release resulting from TPP of DM-nitrophen triggered regenerative Ca2+ release from the neighboring sarcoplasmic reticulum. The present techniques should be useful to investigate the interaction of elementary Ca2+ events, the process leading to global Ca2+ movements, in cardiac myocytes and other types of cells.

Calcium Waves in Skinned Cardiac Myocytes Evoked by Two-Photon Excitation Photolysis of Caged Calcium.