Isolation of and Calcium Kinetics in Cochlear Inner Hair Cells of the Guinea Pig

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Abstract
Single inner hair cells of the guinea pig cochlea were isolated using enzymatic and mechanical techniques. The intracellular free calcium ion concentrations [( Ca2+]i) of the isolated inner hair cells were determined using the Ca2+ sensitive dye fura-2 and digital imaging microscopy. In the presence of 1 micron ionomycin, a Ca2+ ionophore, there was an irreversible increase in [Ca2+]i. The 150 mM KCl stimulation, which induces a depolarization, resulted in a temporary increase in [Ca2+]i. This increase in [Ca2+]i was not observed under conditions of depolarization, in Ca(2+)-free medium. These observations are interpreted to mean that the [Ca2+]i during membrane depolarization mainly originates from an influx of extracellular Ca2+ into the cytoplasm.
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