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TMEM16A/anoctamin1 (ANO1), a calcium (Ca 2+ )-activated chloride (Cl - ) channel, has many functions in various excitable cells and modulates excitability in both Ca 2+ - and voltage-gating modes. However, its gating characteristics and role in primary neural cells remain unclear. Here, we characterized its Ca 2+ - and voltage-dependent components in rod bipolar cells using dissociated and slice preparations of the mouse retina. The I-V curves of Ca 2+ -dependent ANO1 tail current and voltage-gated Ca 2+ channel (VGCC) are similar; as ANO1 is blocked by VGCC inhibitors, ANO1 may be gated by Ca 2+ influx through VGCC. The voltage-dependent component of ANO1 has outward rectifying and sustained characteristics and is clearly isolated by the inhibitory effect of Cl - reduction and T16Ainh-A01, a selective ANO1 inhibitor, in high EGTA, a Ca 2+ chelator. The voltage-dependent component disappears due to VGCC inhibition, suggesting that Ca 2+ is the essential trigger for ANO1. In perforated current-clamping method, the application of T16Ainh-A01 and reduction of Cl - extended excitation periods in rod bipolar cells, revealing that ANO1 induces repolarization during excitation. Overall, ANO1 opens by VGCC activation during physiological excitation of the rod bipolar cell and has a voltage-dependent component. These two gating-modes concurrently provide the intrinsic characteristics of the membrane potential in rod bipolar cells.

Calcium- and Voltage-Dependent Dual Gating ANO1 is an Intrinsic Determinant of Repolarization in Rod Bipolar Cells of the Mouse Retina