Genetic Reagents for Making Split-GAL4 Lines in <i>Drosophila</i>

Dionne, Heather; Hibbard, Karen L; Cavallaro, Amanda; Kao, Jui-Chun; Rubin, Gerald M.

doi:10.1534/genetics.118.300682

Cited by 193 publications

(124 citation statements)

References 22 publications

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“…To construct split-GAL4 lines for potential LC6 target neurons we first selected candidate AD and DBD hemi drivers by visually searching images of GAL4 driver expression patterns (Dionne et al, 2018; Jenett et al, 2012b; Tirian and Dickson, 2017). Typically, we tested several candidate split-GAL4 combinations for each target cell type.…”

Section: Methodsmentioning

confidence: 99%

“…Typically, we tested several candidate split-GAL4 combinations for each target cell type. The AD and DBD hemi drivers we used are from published collections (Dionne et al, 2018; Tirian and Dickson, 2017). A detailed description of split-GAL4 hemi drivers is also available online (https://bdsc.indiana.edu/stocks/gal4/split_intro.html) and the cell-type specific split-GAL4 lines generated in this study together with selected images of their anatomy will be made available at https://www.janelia.org/split-GAL4.…”

Section: Methodsmentioning

confidence: 99%

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Connectivity establishes spatial readout of visual looming in a glomerulus lacking retinotopy

Morimoto

Nern

Zhao

et al. 2020

Preprint

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Visual systems can exploit spatial correlations in the visual scene by using retinotopy, the organizing principle by which neighboring cells encode neighboring spatial locations. However, retinotopy is often lost, such as when visual pathways are integrated with other sensory modalities. How is spatial information processed in the absence of retinotopy? Here, we focused on visual looming responsive LC6 cells in Drosophila, a population whose dendrites collectively tile the visual field, but whose axons form a single glomerulus—a structure lacking retinotopic organization—in the central brain. We identified multiple glomerulus neurons and found that they respond to looming in different portions of the visual field, unexpectedly preserving spatial information. Through EM reconstruction of all LC6 synaptic inputs to the glomerulus, we found that LC6 and downstream cell types form circuits within the glomerulus that establish spatial readout of visual features and contralateral suppression—mechanisms that transform visual information for behavioral control.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Connectivity establishes spatial readout of visual looming in a glomerulus lacking retinotopy

Morimoto

Nern

Zhao

et al. 2020

Preprint

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“…The 4562 Generation 1 GAL4 stocks included in this Phase 1 release (Table S1) were from Jenett, et al (2012) and Tirian & Dickson (2017). We focused on driver lines with available AD or DBD hemidrivers (Tirian & Dickson, 2017;Dionne, et al, 2018). ' R57C10-Flp MCFO' was R57C10-Flp2::PEST in attp18;brp::Snap / CyO;pJFRC201-10XUAS-FRT>STOP>FRT-myr::smGFP-HA in VK00005, pJFRC240-10XUAS-FRT>STOP>FRT-myr::smGFP-V5-THS-10XUAS-FRT>STOP>FRT-myr::smGFP-FLAG in Meissner, et al, 2020 Gen1 MCFO Phase 1 release Page of 10 16…”

Section: Fly Stocksmentioning

confidence: 99%

“…The 4562 listed stocks were crossed to the MCFO reporter and imaged. Available stocks with a corresponding AD or DBD split-GAL4 half (Dionne et al 2018;Tirian and Dickson 2017) were included.…”

Section: Supplementmentioning

confidence: 99%

A searchable image resource ofDrosophilaGAL4-driver expression patterns with single neuron resolution

Meissner

Dorman²,

Nern³

et al. 2020

Preprint

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Precise, repeatable genetic access to specific neurons via the GAL4/UAS system and related methods is a key advantage of Drosophila neuroscience. Neuronal targeting is typically documented using light microscopy of full GAL4 expression patterns, which mostly lack the single-cell resolution required for reliable cell type identification. Here we use stochastic GAL4 labeling with the MultiColor FlpOut approach to generate cellular resolution confocal images at large scale. We are releasing aligned images of 27,000 such adult central nervous systems.An anticipated use of this resource is to bridge the gap between electron microscopyidentified neurons and light microscopy-based intersectional genetic approaches such as the split-GAL4 system. Identifying the individual neurons that make up each GAL4 expression pattern improves the prediction of which GAL4 enhancer fragments best combine via split-GAL4 to target neurons of interest. To this end we have developed the NeuronBridge search tool, which matches these light microscope neuronal images to neurons in the recently published FlyEM hemibrain. This work thus provides a resource and search tool that will significantly enhance both the efficiency and efficacy of split-GAL4 targeting of EM-identified neurons and further advance Drosophila neuroscience.Meissner, et al., 2020Gen1 MCFO Phase 1 release

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“…Cleanly and reliably restricting the expression of the transcriptional activator to molecularly defined cell types in the appropriate time windows is essential to interpret the results of an experiment. Promoter fusions and other methods that utilize regulatory genomic fragments have been used to great effect to generate cell type-specific expression patterns in D. melanogaster, exemplified by large-scale collections generated at institutions like the HHMI Janelia Research Campus that aim to generate reagents that target every neuronal cell type in the fly brain (Dionne et al, 2018;Jenett et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

How to turn an organism into a model organism in 10 ‘easy’ steps

Matthews

Vosshall

2020

Journal of Experimental Biology

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Many of the major biological discoveries of the 20th century were made using just six species: Escherichia coli bacteria, Saccharomyces cerevisiae and Schizosaccharomyces pombe yeast, Caenorhabditis elegans nematodes, Drosophila melanogaster flies and Mus musculus mice. Our molecular understanding of the cell division cycle, embryonic development, biological clocks and metabolism were all obtained through genetic analysis using these species. Yet the 'big 6' did not start out as genetic model organisms (hereafter 'model organisms'), so how did they mature into such powerful systems? First, these model organisms are abundant human commensals: they are the bacteria in our gut, the yeast in our beer and bread, the nematodes in our compost pile, the flies in our kitchen and the mice in our walls. Because of this, they are cheaply, easily and rapidly bred in the laboratory and in addition were amenable to genetic analysis. How and why should we add additional species to this roster? We argue that specialist species will reveal new secrets in important areas of biology and that with modern technological innovations like next-generation sequencing and CRISPR-Cas9 genome editing, the time is ripe to move beyond the big 6. In this review, we chart a 10-step path to this goal, using our own experience with the Aedes aegypti mosquito, which we built into a model organism for neurobiology in one decade. Insights into the biology of this deadly disease vector require that we work with the mosquito itself rather than modeling its biology in another species.

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Genetic Reagents for Making Split-GAL4 Lines in Drosophila

Cited by 193 publications

References 22 publications

Connectivity establishes spatial readout of visual looming in a glomerulus lacking retinotopy

Connectivity establishes spatial readout of visual looming in a glomerulus lacking retinotopy

A searchable image resource ofDrosophilaGAL4-driver expression patterns with single neuron resolution

How to turn an organism into a model organism in 10 ‘easy’ steps

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