Osmoregulatory alterations in myo-inositol uptake by bovine lens epithelial cells. III. Effects of cycloheximide and colchicine on Na⁺-myo-inositol cotransporter activity under hypertonic conditions, inhibition of a plasma membrane osmotic stress protein

Cultured bovine lens epithelial cells adapt to hypertonic sodium stress via an increase in Na⁺-myo-inositol cotransporter activity and accumulate myo-inositol. At least 12 hr of hypertonic exposure was necessary to enhance myo-inositol accumulation; and thereafter, uptake activity continued to increase throughout the duration of a 72-hr exposure period. Switching from hypertonic to isotonic medium for 24 hr reversed the otherwise elevated accumulation activity. The protein synthesis inhibitor, cycloheximide, did not affect myo-inositol uptake in isotonic medium but markedly decreased myo-inositol uptake in hypertonic medium. Cells exposed to hypertonic conditions and the microtubule disrupter, colchicine, similarly showed marked impairment of the otherwise enhanced myo-inositol uptake. These studies indicated that hypertonicity-induced elevation of Na⁺-myo-inositol cotransporter activity in cultured bovine lens epithelial cells is not solely attributed to the increased sodium gradient alone, but rather involves increased de novo synthesis of the Na⁺-myo-inositol cotransporter protein(s).