Alteration of synaptosomal plasma membrane cholesterol content: membrane physical properties and cation transport...

The level of nerve membrane cholesterol was altered by in vitro incubation of rat brain synaptosomal plasma membrane with liposomes having varying cholesterol contents. The normal plasma membrane cholesterol/phospholipid ratio of 0.3-0.4 (mol/mol) could be decreased by about one-half or increased more than 100%. Fluorescence polarization measurements were made using the probe 1,6-diphenyl-1,3,5-hexatriene. At temperatures below 35 percent C, lowering membrane cholesterol led to increased apparent microviscosity, while raising cholesterol content produced little change. However, at 45 percent C a continuous direct relationship existed between experimental membrane cholesterol/phospholipid ratio (ranging from 0.18 to 0.73) and apparent microviscosity. Under standard liposome-synaptosomal plasma membrane exchange conditions, 80% of the initial specific [(3)H]saxitoxin binding activity to the voltage-dependent sodium channel and at least 95% of the (Mg2+,K+)-p-nitrophenylphosphatase activity were preserved. Our results indicate that neither the characteristics of toxin binding nor the kinetics of this enzyme activity is dependent upon membrane cholesterol content.