Cholesterol depletion does not alter the capacitance or Ca handling of the surface or t‐tubule membranes in mouse ventricular myocytes

Cholesterol is a key component of the cell plasma membrane. It has been suggested that the t‐tubule membrane of cardiac ventricular myocytes is enriched in cholesterol and that this plays a role in determining t‐tubule structure and function. We have used methyl‐β‐cyclodextrin (MβCD) to deplete cholesterol in intact and detubulated mouse ventricular myocytes to investigate the contribution of cholesterol to t‐tubule structure, membrane capacitance, and the distribution of Ca flux pathways. Depletion of membrane cholesterol was confirmed using filipin; however, di‐8‐ANEPPS staining showed no differences in t‐tubule structure following MβCD treatment. MβCD treatment had no significant effect on the capacitance:volume relationship of intact myocytes or on the decrease in capacitance:volume caused by detubulation. Similarly, Ca influx and efflux were not altered by MβCD treatment and were reduced by a similar amount following detubulation in untreated and MβCD‐treated cells. These data show that cholesterol depletion has similar effects on the surface and t‐tubule membranes and suggest that cholesterol plays no acute role in determining t‐tubule structure and function. We show that cholesterol depletion using MβCD has no effect on t‐tubule structure or function and does not alter surface or t‐tubular membrane capacitance. These data suggest that the functional differences observed between the surface and t‐tubular membranes are not due to differences in cholesterol content.