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Content archived on 2024-06-10

The interaction between nmda-receptor activation and the polymerization statae of actin in cultured hippocampal neurones

Objective



The level at which signal-transduction is integrated to give Long-term-potentiation (LTP) is unclear, however, recent evidence implicates the cytoskeleton as important. The characteristic ATP-, calcium-dependent rundown of the NMDA-receptor (which is centrally involved in LTP) is reversed by treatments thought to specifically protect the filamentous state of the cytoskeletal protein actin (F-actin) from depolymerization into its monomeric components (G-actin). Since F-actin depolymerization is regulated by calcium-dependent proteases it is thought that the receptor rundown could be through NMDA-receptor activated cytosolic calcium increases. Currently, there are no studies demonstrating a direct effect of NMDA-receptor stimulation on the state of F-actin. This project will directly determine whether such a link exists, and whether treatments thought to alter the polymerisation state of actin are effective. We will use cultured neurones of the rat hippocampus, and the site-selective probes: phalloidin for F-(polymerized), and DNAaseI for G-(depolymerized) actin. Fluorescent conjugates of these probes will allow visualisation of F-, and G-actin simultaneously in Axed, and living neuronal cells. The distribution and intensity of staining will be monitored in single cells using confocal microscopy after systematic treatments.

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Coordinator

Institut National de la Santé et de la Recherche Médicale
EU contribution
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Address
123,Boulevard de Port-Royal
75674 Paris
France

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Participants (1)

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