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> Stages de M2 > Liste des stages proposés pour l’année 2017-2018 > Dégradation de l’AKAP4 et régulation de la mobilité du spermatozoïde

Dégradation de l’AKAP4 et régulation de la mobilité du spermatozoïde

proposé par Nicolas SERGEANT, Inserm UMRS1172 1, place de Verdun 59045 Lille cedex

Projet de stage : Sperm motility is driven by a precise coordination between extracellular and intracellular localisation of transduction signal. A-kinase anchored protein kinases (AKAP) are a family of protein enabling the localization of PKA transduction signal inside different cell-types. AKAP4 is specific to sperm and localize to the principal piece of the flagellum thus enabling the coupling between sperm environment, Calcium / PKC and cAMP / PKA signalling, flagellum motion and egg fertilization. We recently showed that human AKAP4 was proteolysed in the human sperm following a yet undetermined mechanism. This proteolysis process is correlated with motility changes in human sperm. In order, to better understand the mechanism leading to the proteolysis of AKAP4 and therefore regulating AKAP4 activities, we aim to determine the mechanism leading to the degradation of AKAP4. Our hypothesis is that oxidative stress, calcium or AMPc signalling or other environmental factors triggers AKAP4 proteolysis. Using expertise in biochemistry, cell biology as well as newly developed specific monoclonal against N- or C-terminal of AKAP4, signalling pathways regulating Calcium homeostasis or PKA signalling as well as oxydative stress will be investigated in human sperm. We should point out that through their longterm collaboration UMRS1172 and EA4308 have all the biochemical technologies, expertise and facilities for the development of this project.

Techniques mises en œuvre par le stagiaire : Spermogramme, isolation des spermatozoïdes par gradient de densité, culture cellulaire, traitement pharmacologique, électrophorèse SDS-PAGE, western-blot, microscopie à fluorescence.

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