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The effects of ruthenium red (RR) on the cellular functions (intracellular Ca2+ handling and contraction) and permeation of the dye through the cell membrane were investigated in intact ferret papillary muscles. The intracellular Ca2+ concentration ([Ca2+]i), measured using aequorin, was simultaneously recorded with tension. The permeation of the dye through the cell membrane was studied with electronmicroscopy. The preparation was continuously stimulated at 0.2 Hz and treated with 50 microM RR at 30 degrees C. [Ca2+]i was increased by electrical stimulation (0.07 and 2 Hz) and rapid cooling (from 30 to 4 degrees C) (RC). In electrical stimulation, RR time-dependently decreased the peak light of aequorin without a significant change in the time course at 30 degrees C. However, in RC, treatment with RR for about 100 min significantly prolonged the decay time of the light signal and increased the peak light. The peak tension in RC was decreased after treatment with RR for a longer time. The pCa-tension relation of skinned preparations was significantly shifted to the right by 50 microM RR. In the RR (50 microM)-treated specimens, mitochondrial outer membranes were darkly stained if OsO4 was used for fixation. Even though the specimen treated with 500 microM RR was fixed without OsO4 and electron staining, the matrices of mitochondria became electron dense. We concluded that RR could penetrate into intact mammalian cardiac myocytes, and that RR inhibits the release of Ca2+ from the sarcoplasmic reticulum in electrical stimulation, inhibits mitochondrial Ca2+ uptake, and decreases the Ca2+ sensitivity of the myofilaments.

Effects of Ruthenium Red on the Cellular Functions and Ultrastructure in Intact Ferret Ventricular Muscles.