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  Analysis of brain synapse activity and the role of endophilin in its maintenance


   Laboratory of Molecular Biology

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  Dr H McMahon  No more applications being accepted  Competition Funded PhD Project (Students Worldwide)

About the Project

www.endocytosis.org

Endophilin-1 is brain specific and highly concentrated in synapses where it is proposed to be involved in clathrin-mediated endocytosis. We however have recently shown in fibroblast that endophilin-2 marks a non-clathrin dependent pathway of endocytosis, where it is responsible for the internalization of many G-protein coupled receptors and growth-factor receptors. By extension one might propose that the neuronal homologue will have a similar function in synaptic vesicle retrieval.

We culture hippocampal neurons, and have a microscopy setup along with appropriate software to measure and analyse exocytosis of synaptic vesicles in response to electrical depolarization, and their subsequent endocytosis. We also have viral knockdown of protein in place and would now like to analyse the role of proteins like endophilin and other BAR-domain containing proteins (that are enriched in nerve terminals) in vesicle dynamics.

This project will help us understand how the brain works to maintain synaptic function under conditions of light versus intense activity and following various drug treatments, and in animal models of neurodegeneration and synaptic dysfunction. Proteins that cause specific disruption of activity will be further analysed in non-neuronal cell culture models and in vitro, for binding partners and for their mechanisms of action.

Funding Notes

Please see the LMB PhD website for further details:
http://www2.mrc-lmb.cam.ac.uk/students/international-phd-programme/funding/

References

Boucrot, E., …and McMahon, H.T. (2015). Endophilin marks and controls a clathrin-independent endocytic pathway. Nature, 517(7535), 460–465. http://doi.org/10.1038/nature14067

Renard, H.-F., Simunovic, M., Lemière, J., Boucrot, E., Garcia-Castillo, M. D., Arumugam, S., et al. (2015). Endophilin-A2 functions in membrane scission in clathrin-independent endocytosis. Nature, 517(7535), 493–496. http://doi.org/10.1038/nature14064

Boucrot, E., … (McMahon, H. T., & Kozlov, M. M.) (2012). Membrane fission is promoted by insertion of amphipathic helices and is restricted by crescent BAR domains. Cell, 149(1), 124–136. http://doi.org/10.1016/j.cell.2012.01.047

Llobet, A., Gallop, J. L., Burden, J. J. E., Camdere, G., Chandra, P., Vallis, Y., et al. (2011). Endophilin drives the fast mode of vesicle retrieval in a ribbon synapse. The Journal of Neuroscience, 31(23), 8512–8519. http://doi.org/10.1523/JNEUROSCI.6223-09.2011

McMahon, H. T., & Boucrot, E. (2011). Molecular mechanism and physiological functions of clathrin-mediated endocytosis. Nature Reviews. Molecular Cell Biology, 12(8), 517–533. http://doi.org/10.1038/nrm3151

Jao, C. C., Hegde, B. G., Gallop, J. L., Hegde, P. B., McMahon, H. T., Haworth, I. S., & Langen, R. (2010). Roles of amphipathic helices and the bin/amphiphysin/rvs (BAR) domain of endophilin in membrane curvature generation. The Journal of Biological Chemistry, 285(26), 20164–20170. http://doi.org/10.1074/jbc.M110.127811

Gallop, J. L., Butler, P. J. G., & McMahon, H. T. (2005). Endophilin and CtBP/BARS are not acyl transferases in endocytosis or Golgi fission. Nature, 438(7068), 675–678. http://doi.org/10.1038/nature04136

Peter, B. J., …. McMahon, H. T. (2004). BAR domains as sensors of membrane curvature: the amphiphysin BAR structure. Science (New York, N.Y.), 303(5657), 495–499. http://doi.org/10.1126/science.1092586

All these papers can be found on our website: endocytosis.org