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Biophys J, October 1999, p. 1945-1959, Vol. 77, No. 4
*Department of Neurobiology and Behavior and #Howard Hughes Medical Institute, State University of New York at Stony Brook, Stony Brook, New York 11794 USA
Cut-open recordings from Xenopus oocytes
expressing either nerve (PN1) or skeletal muscle (SkM1) Na+
channel 
subunits revealed slow inactivation onset and recovery kinetics of inward current. In contrast, recordings using the macropatch configuration resulted in an immediate negative shift in the
voltage-dependence of inactivation and activation, as well as
time-dependent shifts in kinetics when compared to cut-open recordings.
Specifically, a slow transition from predominantly slow onset and
recovery to exclusively fast onset and fast recovery from inactivation
occurred. The shift to fast inactivation was accelerated by patch
excision and by agents that disrupted microtubule formation.
Application of positive pressure to cell-attached macropatch electrodes
prevented the shift in kinetics, while negative pressure led to an
abrupt shift to fast inactivation. Simultaneous electrophysiological recording and video imaging of the cell-attached patch membrane revealed that the pressure-induced shift to fast inactivation coincided
with rupture of sites of membrane attachment to cytoskeleton. These
findings raise the possibility that the negative shift in voltage-dependence and the fast kinetics observed normally for endogenous Na+ channels involve mechanical destabilization.
Our observation that the 
1 subunit causes similar changes in
function of the Na+ channel subunit suggests that 1
may act through interaction with cytoskeleton.
Biophys J, October 1999, p. 1945-1959, Vol. 77, No. 4
© 1999 by the Biophysical Society 0006-3495/99/10/1945/15 $2.00
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