Physiological and pathological regulation of calcium channel trafficking and function (360G-Wellcome-206279_Z_17_Z)

Voltage-gated CaV2 calcium-channels are essential for presynaptic neurotransmitter release. Knowledge of factors governing the regulation in neurons of N-type (CaV2.2) calcium-channel trafficking and properties is key to understanding their pathological role in neuropathic pain. Both CaV2.2 and the auxiliary subunit alpha2delta-1, which is up-regulated in neuropathic pain models, represent validated pain therapeutic targets. My overarching research aim is to address fundamental questions regarding N-type calcium-channel trafficking and function. For this, we will exploit our key development of a knock-in mouse containing HA-tagged CaV2.2. Firstly, we will analyse expression of CaV2.2 in sensory neurons and spinal cord, and its dysregulation following neuropathic insult. We will examine the role of alpha2delta-1 by making double transgenics with alpha2delta-1 knock-out mice. Secondly, we will dissect the interdependent dual roles of alpha2delta subunits in permitting voltage-dependent activation of CaV2.2 channels and promoting their trafficking, concentrating on the essential requirement for proteolytic maturation of alpha2delta. Finally, we will address the identity and subcellular localisation of the protease involved in alpha2delta processing, since it represents a potential novel therapeutic target in chronic pain. This study will parse out the multiple roles of alpha2delta-subunits in N-type calcium-channel function, and elucidate the molecular mechanism of the alpha2delta-ligand gabapentinoid drugs.