Single-Channel Recordings from Two Types of Amiloride-Sensitive Epithelial Na⁺ Channels

Abstract

We report here the first evidence in intact epithelial cells of unit conductance events from amiloride-sen-sitive Na⁺ channels. The events were observed when patch-clamp recordings were made from the apical surface of cultured epithelial kidney cells (A6). Two types of channels were observed: one with a high selectivity to Na⁺ and one with relatively low selectivity. The characteristics of the low-selectivity channel are as follows: single-channel conductance ranged between 7 and 10 pS (mean = 8.4 ± 1.3), the current-voltage (I–V) relationship displayed little if any nonlinearity over a range of ± 80 mV (with respect to the patch pipette) and the channel Na⁺ /K⁺ selectivity was approximately 3–4:1. Amiloride, a cationic blocker of the channel, reduced channel mean open time and increased channel mean closed times as the voltage of the cell interior was made more negative. Amiloride induced channel flickering at increased negative potentials (intracellular potential with respect to the patch) but did not alter the single-channel conductance or the I–V relationship from that observed in control patches. The characteristics of the high-selectivity channel are: a single-channel conductance of 1–3 pS (mean = 2.8 ± 1.2), the current-voltage relationship is markedly nonlinear with a Na⁺ /K⁺ selectivity greater than 20:1. The mean open and closed times for the two types of channels are quite different, the high-selectivity channel being open only about 10% of the time while the low-selectivity channel is open about 30% of the time.

Key Words:

• kidney cells
• patch clamp
• tissue culture
• kinetic models
• amiloride