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School of Biomedical Sciences, University of Leeds, Leeds LS2 9NQ, United Kingdom
Kawai M, Hussain M, and Orchard CH. Am J Heart Circ Physiol 277: H603-H609, 1999 developed a technique to detubulate rat ventricular myocytes using formamide and showed that detubulation results in a decrease in cell capacitance, Ca2+ current density, and Ca2+ transient amplitude. We have investigated the mechanism of this detubulation and possible direct effects of formamide. Staining ventricular cells with di-8-ANEPPS showed that the t tubule membranes remain inside the cell after detubulation; trapping of FITC-labeled dextran within the t tubules showed that detubulation occurs during formamide washout and that the t tubules appear to reseal within the cell. Detubulation had no effect on the microtubule network but resulted in loss of synchronous Ca2+ release on electrical stimulation. In contrast, formamide treatment of atrial cells did not significantly change cell capacitance, Ca2+ current amplitude, action potential configuration, the Ca2+ transient or the response of the Ca2+ transient to isoprenaline. We conclude that formamide washout induces detubulation of single rat ventricular myocytes, leaving the t tubules within the cell, but without direct effects on cell proteins that might alter cell function.
transverse tubules; rat ventricular cell; di-8-ANEPPS; rat atrial cell; Ca2+ current; Ca2+ transient
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