Alternative splicing of auxiliary β2-subunits stabilizes Cav2.3 Ca²⁺ channel activity in continuously active midbrain dopamine neurons

Abstract

In dopaminergic (DA) substantia nigra (SN) neurons Cav2.3 R-type Ca²⁺-currents contribute to somatodendritic Ca²⁺-oscillations. These may contribute to the selective degeneration of these neurons in Parkinson’s disease (PD) since Cav2.3-knockout is neuroprotective in a PD mouse model. However, the typical Cav2.3 gating would predict complete channel inactivation during SN DA neuronal firing. Here we show that in tsA-201-cells the membrane-anchored β2-splice variants β2a and β2e stabilize Cav2.3 gating properties allowing sustained Cav2.3 availability during simulated pacemaking and enhanced Ca²⁺-currents during bursts. We confirmed the expression of β2a and β2e-subunits in the SN and identified SN DA neurons. Patch-clamp recordings of SN DA neurons in mouse brain slices revealed R-type Ca²⁺-currents similar to β2a- or β2e-stabilized Cav2.3-currents and recordings in cultured murine DA neurons confirmed their activity during pacemaking. Taken together, our data support an important (patho)physiological role of β-subunit alternative splicing for Cav2.3 Ca²⁺-signaling in highly vulnerable SN DA neurons.