Visualization of synaptic specializations in live C. elegans with synaptic vesicle protein-GFP fusions.
Synaptic specializations are difficult to visualize at the light microscope level in living preparations. To circumvent this problem, synaptic vesicle proteins were fused to green fluorescent protein (GFP) and expressed in C. elegans neurons. C. elegans synaptobrevin-GFP and synaptogyrin-GFP fusion proteins were observed to target to synaptic sites. This localization allowed the visualization of synaptic specializations in living animals with light microscopy. Restricted expression of synaptobrevin-GFP fusions in subsets of neurons enables the visualization of individual presynaptic varicosities. The cell biology underlying the sorting of synaptic vesicle proteins, trafficking of vesicles to terminals, and the development of presynaptic specializations is now more amenable to forward genetic analysis using these synaptic markers.
Nonet
M L
ML
Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, MO 63110, USA. nonetm@thalamus.wustl.edu
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans; D018014:Gene Transfer Techniques; D017238:Genes, Helminth; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D016133:Polymerase Chain Reaction / Q000379:methods; D050683:R-SNARE Proteins; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D013569:Synapses / Q000502:physiology* / Q000648:ultrastructure*; D013572:Synaptic Vesicles / Q000502:physiology / Q000648:ultrastructure; D062805:Synaptogyrins
D008164:Luminescent Proteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D050683:R-SNARE Proteins; D011993:Recombinant Fusion Proteins; D062805:Synaptogyrins; D049452:Green Fluorescent Proteins