Reopening of single L-type Ca2+ channels in mouse cerebellar granule cells: dependence on voltage and ion concentration.

1. We recorded the activity of single L-type Ca2+ channels from cell-attached patches on mouse cerebellar granule cells. The experiments investigated the mechanism of channel reopening at negative membrane potentials following a strong depolarization. 2. L-type channels that reopened following a strong depolarization showed a wide distribution of single-channel conductances, which ranged from 16 to 28 pS in the presence of 90 mM Ba2+. 3. The distribution of the latencies before reopening was fitted as the sum of two exponential components with time constants tau f approximately 1 and tau s approximately 12 ms at -70 mV. Hyperpolarization reduced the time constant of the slower component approximately e-fold per 43 mV, but had no effect on the faster component. 4. Raising the concentration of external Ba2+ reduced the time constant of the slower component of the reopening latency without altering the fast component. The time constant of the slow component was approximately 27 ms in 10 mM Ba2+ and decreased to 12 ms in 90 mM Ba2+ at -70 mV. The relation between the time constant and external Ba2+ saturated with an apparent KD of approximately 20 mM. 5. The distribution of reopening times was best fitted as the sum of two exponential components with time constants tau f approximately 0.5 ms and tau s approximately 4.5 ms at -70 mV. The conditional latencies before reopening into either the short or long open state were indistinguishable. 6. The results are consistent with the idea that a positively charged blocker occludes the pore during depolarization and channels reopen as the blocker dissociates following repolarization to negative potentials.