Master 2 and doctorate proposition

Topic

More than 50% of the human spastic paraplegias (HSPs) depend on mutations of four endoplasmic reticulum (ER) resident proteins. Amongst them, are those specific to spastin, a protein with the ability to sever microtubules and therefore to regulate their dynamics. The localization and function of this protein suggest its primary role in determining the shape and distribution of ER and therefore its functions, but to date information is lacking to support these conclusions. Thanks to the different SPAST/SPG4 cell models available in the laboratory we will tackle the question regarding whether the defect in spastin expression or expression of mutated forms of this protein induce ER defects at morphological and functional level. Moreover we will investigate new potential molecular interactions of spastin that may help to understand the physio-pathological role of this protein. The results obtained will be valuable in the identification of possible cellular and molecular targets so as to come up with an effective therapeutic strategy for SPG4 linked-HSP and likely neurodegenerative diseases in which the ER is affected.

Objectives

Investigate the role of spastin in the morphology and dynamics of the ER
Evaluate the level of ER-stress.
Analyze the role of spastin in the homeostasis of calcium concentration.
Validate the interaction of new molecular partners (Y2HS) of spastin in mammalian cells.

Mains methods

  • Fibroblasts and cortical neurons derived from SPG4-KO mice and patients affected by SPAST mutation.
  • Immunofluorescence, time lapse video-imaging, FRAP, calcium measurements.
  • Western-blot, Co-immunoprecipitation.

For any application, send a letter of motivation and a CV to the following email address: andrea.burgo@univ-evry.fr