title: Microtubule-dependent recruitment of Staufen-green fluorescent protein into large RNA-containing granules and subsequent dendritic transport in living hippocampal neurons.
abstract: Dendritic mRNA transport and local translation at individual potentiated synapses may represent an elegant way to form synaptic memory. Recently, we characterized Staufen, a double-stranded RNA-binding protein, in rat hippocampal neurons and showed its presence in large RNA-containing granules, which colocalize with microtubules in dendrites. In this paper, we transiently transfect hippocampal neurons with human Staufen-green fluorescent protein (GFP) and find fluorescent granules in the somatodendritic domain of these cells. Human Stau-GFP granules show the same cellular distribution and size and also contain RNA, as already shown for the endogenous Stau particles. In time-lapse videomicroscopy, we show the bidirectional movement of these Staufen-GFP-labeled granules from the cell body into dendrites and vice versa. The average speed of these particles was 6.4 microm/min with a maximum velocity of 24. 3 microm/min. Moreover, we demonstrate that the observed assembly into granules and their subsequent dendritic movement is microtubule dependent. Taken together, we have characterized a novel, nonvesicular, microtubule-dependent transport pathway involving RNA-containing granules with Staufen as a core component. This is the first demonstration in living neurons of movement of an essential protein constituent of the mRNA transport machinery.
authors.affiliation: European Molecular Biology Laboratory, Cell Biology Programme, 69117 Heidelberg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Köhrmann
authors.forename: M
authors.initials: M
authors.lastname: Luo
authors.forename: C
authors.initials: C
authors.lastname: Kaether
authors.forename: L
authors.initials: L
authors.lastname: DesGroseillers
authors.forename: C G
authors.initials: CG
authors.lastname: Dotti
authors.forename: M A
authors.initials: MA
authors.lastname: Kiebler
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins; D003712:Dendrites / Q000187:drug effects / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D006624:Hippocampus / Q000166:cytology* / Q000196:embryology; D006801:Humans; D007700:Kinetics; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism*; D009068:Movement / Q000187:drug effects; D015739:Nocodazole / Q000494:pharmacology; D016011:Normal Distribution; D015388:Organelles / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D003598:Cytoskeletal Proteins; D008164:Luminescent Proteins; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C492185:STAU1 protein, human; C492186:Stau1 protein, rat; C117884:staufen protein, mammalian; D049452:Green Fluorescent Proteins; D012313:RNA; D015739:Nocodazole