Mechanically gated channel activity in cytoskeleton-deficient plasma membrane blebs and vesicles from Xenopus oocytes

1. A novel technique involving hypertonic stress causes membrane 'blebbing' of the Xenopus oocyte and the shedding of plasma membrane vesicles (PMVs). 2. Confocal fluorescence microscopy, immunocytochemistry and electron micro scopy indicate that blebs and PMVs lack cortical cytoskeleton and are defic ient in cytoskeleton proteins and devoid of microvilli. 3. Patch recordings from PMVs consistently reveal mechanically gated (MG) c hannel activity. The MG channels display the same single-channel conductanc e as control recordings but differ in terms of reduced mechanosensitivity a nd adaptation to sustained stimulation. 4. Whole PMV recordings show rapid and reversible activation of mechanosens itive currents in response to pressure pulses. The maximal currents activat ed in PMVs are consistent with MG channel activity recorded in patches. 5. The discrepancy between MG channel activity recorded in whole PMVs and o ocytes most probably reflects their different membrane geometry and ability to develop activating bilayer tensions. 6. We propose that membrane blebbing, which is known to occur under specifi c physiological and pathological conditions (e.g. mitosis and apoptosis), m ay increase mechanosensitivity independently of the intrinsic properties of membrane proteins.