Contributions of Ca_V1.3 Channels to Ca²⁺ Current and Ca²⁺-Activated BK Current in the Suprachiasmatic Nucleus

Daily regulation of Ca²⁺ - and voltage-activated BK K⁺ channel activity is required for action potential rhythmicity in the suprachiasmatic nucleus (SCN) of the hypothalamus, the brain's circadian clock. In SCN neurons, BK activation is dependent upon multiple types of Ca²⁺ channels in a circadian manner. Daytime BK current predominantly requires Ca²⁺ influx through L-type Ca²⁺ channels (LTCCs), a time when BK channels are closely coupled with their Ca²⁺ source. Here we show that daytime BK current is resistant to the Ca²⁺ chelator BAPTA. However, at night when LTCCs contribute little to BK activation, BK current decreases by a third in BAPTA compared to control EGTA conditions. In phase with this time-of-day specific effect on BK current activation, LTCC current is larger during the day. The specific Ca²⁺ channel subtypes underlying the LTCC current in SCN, as well as the subtypes contributing the Ca²⁺ influx relevant for BK current activation, have not been identified. SCN neurons express two LTCC subtypes, Ca_V1.2 and Ca_V1.3. While a role for Ca_V1.2 channels has been identified during the night, Ca_V1.3 channel modulation has also been suggested to contribute to daytime SCN action potential activity, as well as subthreshold Ca²⁺ oscillations. Here we characterize the role of Ca_V1.3 channels in LTCC and BK current activation in SCN neurons using a global deletion of CACNA1D in mouse (Ca_V1.3 KO). Ca_V1.3 KO SCN neurons had a 50% reduction in the daytime LTCC current, but not total Ca²⁺ current, with no difference in Ca²⁺ current levels at night. During the day, Ca_V1.3 KO neurons exhibited oscillations in membrane potential, and most neurons, although not all, also had BK currents. Changes in BK current activation were only detectable at the highest voltage tested. These data show that while Ca_V1.3 channels contribute to the daytime Ca²⁺ current, this does not translate into a major effect on the daytime BK current. These data suggest that BK current activation does not absolutely require Ca_V1.3 channels and may therefore also depend on other LTCC subtypes, such as Ca_V1.2.