Cardiac t-tubules undergo significant remodeling in various pathological and experimental conditions, which can be associated with mechanical or osmotic stress. In particular, it has been shown that removal of hyposmotic stress can lead to sealing of t-tubules. However, the mechanisms underlying the sealing process remain essentially unknown. In this study we use dextran trapping assay to demonstrate that in adult mouse cardiomyocytes t-tubular sealing can also be induced by hyperosmotic challenge, and that both hypo- and hyperosmotic sealing display a clear threshold behavior requiring ≈100 mOsm minimal stress. Importantly, during both hypo- and hyperosmotic challenges the sealing of t-tubules occurs only during the shrinking phase. Analysis of the time course of t-tubular remodeling following removal of hyposmotic stress shows that t-tubules become sealed essentially instantly, well before any significant reduction in cell size can be observed. Overall, the data support the hypothesis that the critical event in the process of t-tubular sealing during osmotic challenges is detachment (peeling) of the membrane from the underlying cytoskeleton due to suprathreshold stress.
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