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Localizing Protein in 3D Neural Stem Cell Culture: a Hybrid Visualization Methodology
Published on: December 19, 2010
Subcellular localization of neural-specific NPDC-1 protein
1Laboratoire Biologie Moléculaire et Différenciation, Unité de Génétique Oncologique, CNRS-URA 8125, Institut Gustave Roussy, Villejuif, France. clevrard@free.fr
Abstract:
NPDC-1 is a gene specifically expressed in neural cells when they stop to divide and begin to differentiate. Immunocytochemical study analysis of differentiated PC12 cells transfected with NPDC-tag vectors showed that NPDC-1 is transported in vesicles from the Golgi apparatus to the cell membrane and is then likely internalized into endosomes. The protein colocalized, at least partially, with synaptic vesicle proteins: synaptophysin, synaptobrevin 2, and Rab3 GEP (Rab3 GTP/GDP exchange protein). Moreover, subcellular fractionation of rat brain showed that crude synaptic membrane and crude synaptic vesicle fractions were enriched in NPDC-1. Although NPDC-1 bound Rab3 GEP in vitro, it seems unlikely to be involved in Ca2+-dependent exocytosis and, thus, in synaptic vesicle trafficking.
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