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cytoskeletal dyamics
Currently viewing: 24 September - 4 October 2007 | Gif-sur-Yvette | FR

Preliminary programme

The course will begin on Monday, 24 September in the morning and last until Thursday, 4 October in the afternoon. Ten working days will be dedicated to the course and Sunday, 30 September will be a day off.

Each day will start with an introductory tutorial seminar. Most of the day will be devoted to practical teaching and will end with a lecture given by an instructor. After dinner will be devoted to free discussions. Computers will be available for participants and lecturers to share additional data, movies, simulations and to entertain discussions. Free space will be available for posters if volunteered by participants. For a brief overview of the program, applicants can refer to the course titles indicated in the list of instructors.

This course will cover the following main topics:

  1. Actin/tubulin assembly dynamics in solution in vitro:
    kinetic and steady state measurements of actin assembly by fluorescence spectroscopy and light scattering will be used for the functional characterisation of regulatory proteins, with examples like G-actin sequestering proteins, barbed end cappers, leaky cappers, profilin function, filament branching enzymes and nucleators.

  2. Visual assay for individual polymer dynamics in vitro:
    real time imaging of individual actin filament or microtubule assembly by fluorescence microscopy (TIRF or evanescent wave microscopy) will be used to observe simultaneously in double fluorescence:
    1. actin filaments branching and the localisation of the branching agent Arp2/3
    2. assembly of actin filaments and the processive motor formin
    3. disassembly of microtubules and a processive motor like a kinesin.

  3. Reconstituted biomimetic systems:
    real-time imaging of actin-based propulsive movement of N-WASP or formin-coated beads or liposomes, reconstitution of actin assembly from integrin-adhesion complexes, liposome constriction by endocytic proteins will be observed in phase contrast and (single or double) fluorescence microscopy. Image and movie analysis.

  4. Interface between membrane and cytoskeleton, measurement of the force produced by polymer growth against a membrane, in transport processes: force produced by a growing microtubule, growth of tubules mediated by dynamin, or by kinesin along microtubules; real-time imaging of the diffusion of machineries initiating branched filament networks in a functionalised liposome using FRAP.

  5. Observation of motile processes in a living cell:
    Dictyostelium discoideum will be used to monitor chemotactic movement, filopodia formation; fission yeast will be used for live cell microscopy analysis of mitosis and cytokinetic ring assembly; use of Confocal microscopy, FRAP measurements of turnover of actin filaments.

  6. Modelling of cytoskeletal dynamics in cellular processes:
    Computer simulations of the dynamics of cytoskeletal polymers based on classical mechanics and calculation of forces present in the molecular assemblies.

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