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Large-scale study of phosphoproteins involved in long-term potentiation in the rat dentate gyrus in vivo
Authors:Chardonnet Solenne  Le Marechal Pierre  Cheval Hélène  Le Caer Jean-Pierre  Decottignies Paulette  Laprevote Olivier  Laroche Serge  Davis Sabrina
Affiliation:Laboratoire de Neurobiologie de l'Apprentissage, de la Mémoire et de la Communication, UniversitéParis-Sud, Orsay, France;
CNRS, UMR 8620, Orsay, France;
Institut de Biochimie et Biophysique Moléculaire et Cellulaire, Chimie des protéines, Bâtiment 430, UniversitéParis-Sud, 91405 Orsay Cedex, France;
CNRS, UMR 8619, Orsay, France;
ICSN, UPR2301, CNRS, Gif-sur-Yvette, France
Abstract:The mechanisms underlying the induction of synaptic plasticity and the formation of long-term memory involve activation of cell-signalling cascades and protein modifications such as phosphorylation and dephosphorylation. Based on a protein candidate strategy, studies have identified several protein kinases and their substrates, which show an altered phosphorylation state during the early phases of long-term potentiation (LTP), yet only a limited number of synaptic phosphoproteins are known to be implicated in LTP. To identify new phosphoproteins associated with LTP, we have undertaken a proteomic study of phosphoproteins at different time points following the induction of LTP in the dentate gyrus in vivo (0, 15 and 90 min). For each time point, proteins from the dentate gyrus were separated by two-dimensional gel electrophoresis and stained with Pro−Q® Diamond, a fluorescent stain specific for phosphoproteins. Fourteen proteins whose phosphorylation state varied significantly following LTP were identified using matrix-assisted laser desorption ionization/time of flight mass spectrometry and electrospray ionization-Orbitrap tandem mass spectrometry (MS/MS). They are involved in various cellular functions implicated in synaptic plasticity, such as intracellular signalling, axonal growth, exocytosis, protein synthesis and metabolism. Our results highlight new proteins whose phosphorylation or dephosphorylation is associated with LTP induction or maintenance. Further studies focusing on the regulation of specific phosphorylation sites will lead to greater understanding of the individual implications of these proteins in LTP as well as of their molecular interactions.
Keywords:hippocampus    long-term potentiation    phosphorylation    proteomics    synaptic plasticity
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