Visualization of synaptic specializations in live C. elegans with synaptic vesicle protein-GFP fusions

Michael L. Nonet

doi:10.1016/S0165-0270(99)00031-X

Visualization of synaptic specializations in live C. elegans with synaptic vesicle protein-GFP fusions

Michael L. Nonet

Research output: Contribution to journal › Article › peer-review

189 Scopus citations

Abstract

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. Copyright (C) 1999 Elsevier Science B.V.

Original language	English
Pages (from-to)	33-40
Number of pages	8
Journal	Journal of Neuroscience Methods
Volume	89
Issue number	1
DOIs	https://doi.org/10.1016/S0165-0270(99)00031-X
State	Published - Jul 1 1999

Keywords

AP180
Fluorescence microscopy
Green fluorescent protein
In vivo imaging
Synapse
Synaptic vesicle
Synaptobrevin
Synaptogyrin

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10.1016/S0165-0270(99)00031-X

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title = "Visualization of synaptic specializations in live C. elegans with synaptic vesicle protein-GFP fusions",

abstract = "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. Copyright (C) 1999 Elsevier Science B.V.",

keywords = "AP180, Fluorescence microscopy, Green fluorescent protein, In vivo imaging, Synapse, Synaptic vesicle, Synaptobrevin, Synaptogyrin",

author = "Nonet, {Michael L.}",

note = "Funding Information: I thank A. Fire, S. White-Harrison, D. Dixon, J. Ahnn, G. Seydoux and S. Xu for the GFP vectors pPD114.42, pPD95.75, pPD95.79 and pPD52.102, Aixa Alfonso for pFB11, David Miller for providing the unc-4 promoter, Anna Borodovski for pAB8, Owais Saifee for pTX41, Anneliese M. Schaefer for examining L1 jsIs37 animals, Liping Wei for aiding in integrating transgenes, and Yishi Jin, Maya Kunkel, David Miller, Aguan Wei and members of my laboratory for critical comments. This work was supported by the Beckman, Searle and McKnight foundations.",

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doi = "10.1016/S0165-0270(99)00031-X",

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AU - Nonet, Michael L.

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N2 - 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. Copyright (C) 1999 Elsevier Science B.V.

AB - 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. Copyright (C) 1999 Elsevier Science B.V.

KW - AP180

KW - Fluorescence microscopy

KW - Green fluorescent protein

KW - In vivo imaging

KW - Synapse

KW - Synaptic vesicle

KW - Synaptobrevin

KW - Synaptogyrin

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EP - 40

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

IS - 1

ER -

Visualization of synaptic specializations in live C. elegans with synaptic vesicle protein-GFP fusions

Abstract

Keywords

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