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Cardiovascular Research 1992 26(2):179-185; doi:10.1093/cvr/26.2.179
© 1992 by European Society of Cardiology
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Copyright © 1992, European Society of Cardiology

Effects of isoprenaline and ouabain on cytosolic calcium and cell motion in single rat cardiomyocytes

Kazuhiko Tamura, Shohshin Yoshida, Takeshi Iwai and Ichiro Watanabe

Research Laboratories, Chugai Pharmaceutical Co Ltd, 135 Komakado 1 chome, Gotemba-shi, Shizuoka 412, Japan: K Tamura, S Yoshida, T Iwai, I Wantabe.

Correspondence to Dr Tamura.

Objective: The aim was to develop a novel system to measure changes in cytosolic free calcium concentration ([Ca2+]i) and cell motion simultaneously in a single beating myocyte by digital imaging microscopy. Methods: Indo-1 was selected as a fluorescent calcium indicator. The particular feature of our digital imaging microscopy is that, in order to avoid cell movement artifacts, two indo-1 fluorescence images of different wavelengths are obtained at the same time at 16.7 ms intervals using two SIT cameras. Isolated ventricular myocytes were used, prepared from hearts of male Sprague-Dawley rats, weighing 350 to 400 g. Results: In continuously beating cells contracted by electrical field stimulation, we can visualise changes in [Ca2+]i and cell shape during contraction with pseudocolour images of the fluorescence ratios, and it was found that the onset of the calcium transient preceded that of the shortening of cell length. Both isoprenaline and ouabain enhanced the magnitudes of the calcium transient and cell contraction. Furthermore, isoprenaline shortened the half decay time of the calcium transient. Conclusions: Our data suggest that the newly developed system is useful for measuring rapid changes in [Ca2+]i in relation to cell motion, and is particularly suitable for measuring the calcium transients in a beating cardiomyocyte.

KEYWORDS digital image processing; intracellular Ca2+; concentration; indo-1; cardiac myocytes; isoprenaline; ouabain


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