Abstract
Excised patches of oocyte membrane containing cx46 hemichannels were used to determine the accessibility of the channels for several uncharged molecules at negative and positive holding potentials. The molecular weights of the test molecules (sugars and polyethylene glycols) ranged from 180 to 666 Daltons, with diameters from 5.8 to 12 A. Activation of the voltage gate (V(j) gate) at positive potentials shifted the accessibility limit for these test molecules to lower sizes indicating that net charge of the test molecule does not determine accessibility. The sugars changed several channel properties: 1), single-channel conductance decreased in an inverse relationship to the size of the test molecule; 2), the apparent open probability of the connexin channels was reduced; 3), the lifetimes of the apparent occupancy states exceeded the expected transit times assuming simple diffusion by orders of magnitude. These results suggest a channel that is not cylindrical and in which test molecules bind or are trapped in the pore or the vestibulum. Furthermore, the effect of sugars and the large difference in transit rates for small ions and larger molecules suggest that they may involve different permeation mechanisms.