Expression of Rab3A GTPase and other synaptic proteins is induced in differentiated NT2N neurons.
Postmitotic NT2N cells, which are derived from human NT2 teratocarcinoma cells by treatment with retinoic acid (RA) and mitotic inhibitors, are viewed as a good in vitro model for mature neurons of the human central nervous system. Although NT2N cells exhibit many morphological and biochemical characteristics of neurons, the expression of key protein components involved in regulated exocytosis have not been firmly established. Here we show by immunoblot analysis that mature morphologically differentiated NT2N cells contain readily detectable quantities of the synaptic vesicle-associated proteins, synaptobrevin, synapsin, and synaptophysin. They also express the presynaptic plasma membrane protein, SNAP-25, and a Rab GTPase implicated in the control of Ca(2+)-dependent exocytosis, Rab3A. These proteins were not detected in untreated NT2 cells or cells exposed to RA for only 6 d. The induction of an array of proteins known to be involved in the docking and fusion of synaptic vesicles with the plasma membrane provides further support for the validity of NT2N cells as a model for human cortical neurons and suggests that these cells may be useful for in vitro molecular studies of the Ca(2+)-regulated exocytic pathway in nerve terminals.
Hood Research Program, Weis Center for Research, Pennsylvania State University College of Medicine, Danville 17822-2616, USA.
K M
KM
Sheridan
W A
WA
Maltese
D014408:Biomarkers, Tumor; D002454:Cell Differentiation; D020558:GTP Phosphohydrolases / Q000096:biosynthesis*; D019204:GTP-Binding Proteins / Q000096:biosynthesis*; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis; D009419:Nerve Tissue Proteins / Q000096:biosynthesis*; D009474:Neurons / Q000378:metabolism*; D050683:R-SNARE Proteins; D016704:Synapsins / Q000096:biosynthesis; D013572:Synaptic Vesicles / Q000378:metabolism; D016708:Synaptophysin / Q000096:biosynthesis; D050825:Synaptosomal-Associated Protein 25; D018243:Teratocarcinoma / Q000378:metabolism; D013736:Testicular Neoplasms / Q000378:metabolism; D014407:Tumor Cells, Cultured; D020692:rab3 GTP-Binding Proteins
D014408:Biomarkers, Tumor; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D050683:R-SNARE Proteins; C494208:SNAP25 protein, human; D016704:Synapsins; D016708:Synaptophysin; D050825:Synaptosomal-Associated Protein 25; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins; D020692:rab3 GTP-Binding Proteins