Abstract
Two kinds of single channels, carrying inward currents even above the Na and Cl ion equilibrium potentials, were observed in outside-out patches from Aplysia neurones bathed in K-free internal and external solutions. The channel carrying the larger elementary current has been studied in detail. When the internal solution contained mainly CsCl, this channel usually inactivated during the first minutes following isolation of the membrane patch. However, when the internal solution contained NaCl instead of CsCl, the channel remained functional during several hours, thus allowing the present study. Na-Tris, NaCl-mannitol and Ca-Ba external substitution experiments showed that the channel studied is much more permeable to divalent cations than to sodium ions. Mono-exponential open-time distributions obtained under identical conditions from different membrane patches indicated either slow (in the order of 100 ms at 0 mV) or rapid (a few milliseconds at 0 mV) mean open-times. Biphasic open-time distributions could be obtained from other membrane patches under the same conditions. These results suggest the existence of two different gating modes. Both the open-time distribution and the closed-time distribution are voltage sensitive: membrane depolarization activates the channel by lengthening the openings and shortening the closures. The threshold of activation if any, is very low and the inactivation, if present, is never complete. Ca-Ba, Ca-Sr and Ca-Mg external substitution experiments showed that the elementary current amplitude is not very sensitive to the nature of the external divalent cation. The elementary current can be slightly larger when carried by Ba ions rather than by Ca ions, but is nearly identical whether carried by Ca, Sr or even Mg ions, which leads to the elementary conductance sequence: Ba greater than or equal to Ca = Sr congruent to Mg. In contrast, the mean open-time of the channel is very sensitive to the nature of the external permeant ion. The longest mean open-time is observed in the presence of Ca ions, and the mean open-time sequence is: Ca greater than Sr greater than Ba greater than Mg. The closed-time distribution is also affected by the nature of the external divalent cation. The above results show that the nature of the permeant ion affects the kinetic properties of the channel much more than its elementary current.(ABSTRACT TRUNCATED AT 400 WORDS)