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Conditional Synaptic Vesicle Markers for Drosophila.

Jessica L Williams | Harold K Shearin | R Steven Stowers
G3 (Bethesda, Md.) | 2019

The release of neurotransmitters from synaptic vesicles (SVs) at pre-synaptic release sites is the principle means by which information transfer between neurons occurs. Knowledge of the location of SVs within a neuron can thus provide valuable clues about the location of neurotransmitter release within a neuron and the downstream neurons to which a given neuron is connected, important information for understanding how neural circuits generate behavior. Here the development and characterization of four conditional tagged SV markers for Drosophila melanogaster is presented. This characterization includes evaluation of conditionality, specificity for SV localization, and sensitivity of detection in diverse neuron subtypes. These four SV markers are genome-edited variants of the synaptic vesicle-specific protein Rab3. They depend on either the B2 or FLP recombinases for conditionality, and incorporate GFP or mCherry fluorescent proteins, or FLAG or HA epitope tags, for detection.

Pubmed ID: 30635441

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Addgene (tool)

RRID:SCR_002037

Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.

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New England Biolabs (tool)

RRID:SCR_013517

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w[*]; P{w[+mC]=UAS-mCD8.ChRFP}3 (organism)

RRID:BDSC_27392

Drosophila melanogaster with name w[*]; P{w[+mC]=UAS-mCD8.ChRFP}3 from BDSC.

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w[*]; P{w[+mC]=UAS-FLP.D}JD2 (organism)

RRID:BDSC_4540

Drosophila melanogaster with name w[*]; P{w[+mC]=UAS-FLP.D}JD2 from BDSC.

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w[*]; Rab3[rup] (organism)

RRID:BDSC_78045

Drosophila melanogaster with name w[*]; Rab3[rup] from BDSC.

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w[*]; P{w[+mC]=UAS-syt.eGFP}3 (organism)

RRID:BDSC_6926

Drosophila melanogaster with name w[*]; P{w[+mC]=UAS-syt.eGFP}3 from BDSC.

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w[*]; P{w[+mC]=UAS-nSyb.eGFP}3 (organism)

RRID:BDSC_6922

Drosophila melanogaster with name w[*]; P{w[+mC]=UAS-nSyb.eGFP}3 from BDSC.

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w[1118]; TI{TI}VAChT[FRT-STOP-FRT.HA]/TM6B, Tb[1] (organism)

RRID:BDSC_76021

Drosophila melanogaster with name w[1118]; TI{TI}VAChT[FRT-STOP-FRT.HA]/TM6B, Tb[1] from BDSC.

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