Rapid adaptation of single mechanosensitive channels in Xenopus oocytes.
Mechanosensitive (MS) channels are expressed in a wide range of cell types and have been implicated in diverse functions, including osmoregulation and mechanoreception. The majority of previous studies on single MS channels have been carried out on nonsensory cells and have dealt with the steady-state properties of the channel. Here we measure the dynamic or nonstationary properties of the MS channel in Xenopus laevis oocytes. MS channels open transiently in response to a step change in suction applied to the membrane patch. This adaptive behavior occurs because of a reduction in open channel probability rather than a decrease in channel conductance. Double-step suction protocols indicate that adapted MS channels can be reactivated by application of stronger stimulation, consistent with a change in gating sensitivity rather than channel inactivation. Adaptation is highly voltage dependent, being most evident at resting or hyperpolarized potentials and absent at strongly positive potentials. Neither adaptation nor its voltage sensitivity requires the presence of extracellular Ca2+. Adaptation is fragile, dependent on patch history, and can be irreversibly abolished by moderate suction applied to the patch while MS channel activity is retained. Further suction can abolish MS channel activity without compromising the seal. We propose that the selective loss of adaptation and MS channel activity is due to different stages of membrane-cytoskeleton decoupling caused by the mechanical stresses associated with patch clamp recording.
Hamill
O P
OP
Section of Neurobiology and Behavior, Cornell University, Ithaca, NY 14853.
McBride
D W
DW
D000064:Acclimatization; D000818:Animals; D004533:Egtazic Acid / Q000494:pharmacology; D005260:Female; D015640:Ion Channel Gating; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology*; D008465:Mechanoreceptors / Q000187:drug effects / Q000502:physiology*; D008564:Membrane Potentials / Q000187:drug effects; D009865:Oocytes / Q000502:physiology*; D014982:Xenopus laevis
D007473:Ion Channels; D004533:Egtazic Acid