4 YEAR PhD IN NEUROSCIENCE
Department of Neuroscience, Physiology & Pharmacology
Voltage-dependent calcium channels
- The voltage gated calcium channels consist of a pore-forming α1 subunit with b and a2δ auxiliary subunits.
- In my lab we work on several main areas including the role of the calcium channel α2δ subunits and their relation to disease.
- The α2δ-1 and α2δ-2 proteins are therapeutic targets for the widely used anti-epileptic drug gabapentin, which is also effective in various neuropathic pain conditions. Furthermore α2δ-1 is up-regulated in damaged sensory neurons in experimental models of neuropathic pain.
- The projects relate to this area, which is funded by a Wellcome Trust Senior Investigator award.
Three month project placement for Wellcome Neuroscience PhD students.
Example Project: Investigation of synaptic expression of calcium channel subunits in neurons. Hypothesis Voltage-gated calcium channel a2δ subunits traffic N type calcium channels (CaV2.2) to synapses Aims We have made CaV2.2 with a tag in an external loop that enables the proteins to be labelled directly with a fluorophore. We aim to express this in neurons together with other proteins, including other calcium channel subunits and fluorescently labelled synaptic markers. We will use a number of types of neuron, for example hippocampal neurons or dorsal root ganglion neurons Methods The methods used will involve molecular biology, confocal imaging and analysis. You will also learn to culture neurons for expression studies by and transfect by nucleofection. If the project were extended to a PhD you would perform live cell imaging studies in neurons and examine the effect of gabapentin on the ability of α2δ subunits to traffic calcium channels.
Hoppa M, Lana B, Margas W, Dolphin AC, Ryan TA (2012) a2δ couples calcium channels to neurotransmitter release sites to control release probability. Nature 486:122-125.
A, Kadurin I, Alvarez-Laviada A, Douglas L, Bauer CS, Nieto-Rostro M, Pratt, WS
and Dolphin AC (2010) The α2δ subunits of voltage-gated calcium channels form
GPI-anchored proteins, a post-translational modification essential for
function. Proc Natl Acad Sci U S A.
Bauer CS, Nieto-Rostro M, Rahmin W, Tran-Van-Minh A, Ferron L, Douglas L, Kadurin, I, Sri Ranjan Y, Fernandez-Alacid L, Millar NS, Dickenson AH, Lujan R and Dolphin AC (2009).. Journal of Neuroscience 29:4076–4088
Hendrich J, Tran Van Minh A, Heblich F, Nieto-Rostro M, Watschinger K, Striessnig J, Wratten J, Davies A and Dolphin AC (2008) Pharmacological disruption of calcium channel trafficking by the α2δ ligand gabapentin. Proc Natl Acad Sci U S A. 105: 3628–3633.
Field,M.J., Cox,P.J., Stott,E., Melrose,H., Offord,J., Su,T.Z., Bramwell,S., Corradini,L., England,S., Winks,J., Kinloch,R.A., Hendrich,J., Dolphin,A.C., Webb,T., and Williams,D (2006) Identification of the α2δ-1 subunit of voltage-dependent calcium channels as a novel molecular target for pain mediating the analgesic actions of pregabalin. Proc. Natl. Acad. Sci. (USA) 103, 17537-17542
Canti C, Nieto-Rostro M, Foucault I, Heblich F, Wratten J, Richards MW, Hendrich J, Douglas L, Page KM, Davies A, Dolphin AC (2005) The metal-ion-dependent adhesion site in the Von Willebrand factor-A domain of α2δ subunits is key to trafficking voltage-gated Ca2+ channels. Proc Natl Acad Sci U S A. 102: 11230-11235
Viard P, Butcher A, Halet G, Davies A, Nürnberg B, Heblich F and Dolphin AC (2004) PI3K promotes voltage-dependent calcium channel trafficking to the plasma membrane. Nature Neuroscience 7: 939-946