A GAL4-Driver Line Resource for Drosophila Neurobiology

Cell Reports, Journal Year: 2012, Volume and Issue: 2(4), P. 991 - 1001

Published: Oct. 1, 2012

We established a collection of 7,000 transgenic lines Drosophila melanogaster. Expression GAL4 in each line is controlled by different, defined fragment genomic DNA that serves as transcriptional enhancer. used confocal microscopy dissected nervous systems to determine the expression patterns driven adult brain and ventral nerve cord. present image data on 6,650 lines. Using both manual machine-assisted annotation, we describe most useful illustrate utility these for identifying novel neuronal cell types, revealing asymmetry, describing nature extent shape stereotypy. The allow exogenous genes distinct, small subsets system. set fragments, driving documented pattern, will facilitate generation additional constructs manipulating function.

Language: Английский

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Mushroom body output neurons encode valence and guide memory-based action selection in Drosophila

eLife, Journal Year: 2014, Volume and Issue: 3

Published: Dec. 22, 2014

Animals discriminate stimuli, learn their predictive value and use this knowledge to modify behavior. In Drosophila, the mushroom body (MB) plays a key role in these processes. Sensory stimuli are sparsely represented by ∼2000 Kenyon cells, which converge onto 34 output neurons (MBONs) of 21 types. We studied MBONs several associative learning tasks sleep regulation, revealing extent information flow is segregated into distinct channels suggesting possible roles for multi-layered MBON network. also show that optogenetic activation can, depending on cell type, induce repulsion or attraction flies. The behavioral effects perturbation combinatorial, ensemble collectively represents valence. propose local, stimulus-specific dopaminergic modulation selectively alters balance within network those stimuli. Our results suggest valence encoded biases memory-based action selection.

Language: Английский

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Optimized tools for multicolor stochastic labeling reveal diverse stereotyped cell arrangements in the fly visual system

Proceedings of the National Academy of Sciences, Journal Year: 2015, Volume and Issue: 112(22)

Published: May 11, 2015

Significance Nervous systems contain vast numbers of neurons with diverse shapes and complex spatial relationships. We describe new genetic tools for the efficient visualization by light microscopy individual their relative positions in Drosophila . The application these methods to visual system revealed an unexpected diversity cell-type–specific arrangements neuronal processes within a single brain region. This wide range stereotyped cell provides distinct circuit elements processing information implies existence surprisingly large number programs that produce during development.

Language: Английский

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A Framework for Studying Emotions across Species

Cell, Journal Year: 2014, Volume and Issue: 157(1), P. 187 - 200

Published: March 1, 2014

Language: Английский

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Drosophilatools and assays for the study of human diseases

Disease Models & Mechanisms, Journal Year: 2016, Volume and Issue: 9(3), P. 235 - 244

Published: March 1, 2016

ABSTRACT Many of the internal organ systems Drosophila melanogaster are functionally analogous to those in vertebrates, including humans. Although humans and flies differ greatly terms their gross morphological cellular features, many molecular mechanisms that govern development drive physiological processes conserved between both organisms. The differences deceiving have led researchers undervalue study invertebrate organs unraveling pathogenic diseases. In this review accompanying poster, we highlight parallels fly human validate use pathogenesis underlying We discuss assays been developed function specific genes central nervous system, heart, liver kidney, provide examples these address questions related These us with simple yet powerful tools associated disease-causing genes.

Language: Английский

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A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila

eLife, Journal Year: 2015, Volume and Issue: 4

Published: March 31, 2015

Here, we document a collection of ∼7434 MiMIC (Minos Mediated Integration Cassette) insertions which 2854 are inserted in coding introns. They allowed us to create library 400 GFP-tagged genes. We show that 72% internally tagged proteins functional, and more than 90% can be imaged unfixed tissues. Moreover, the mRNAs knocked down by RNAi against GFP (iGFPi), efficiently deGradFP technology. The phenotypes associated with RNA protein knockdown typically correspond severe loss function or null mutant phenotypes. Finally, demonstrate reversible, spatial, temporal larvae adult flies. This new strategy strains allows unprecedented vivo manipulations flies for many These strategies will likely extend vertebrates.

Language: Английский

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Plug-and-Play Genetic Access to Drosophila Cell Types using Exchangeable Exon Cassettes

Cell Reports, Journal Year: 2015, Volume and Issue: 10(8), P. 1410 - 1421

Published: Feb. 26, 2015

Genetically encoded effectors are important tools for probing cellular function in living animals, but improved methods directing their expression to specific cell types required. Here, we introduce a simple, versatile method achieving cell-type-specific of transgenes that leverages the untapped potential "coding introns" (i.e., introns between coding exons). Our couples transgene native gene expressed cells interest using intronically inserted "plug-and-play" cassettes (called "Trojan exons") carry splice acceptor site followed by sequences T2A peptide and an effector transgene. We demonstrate efficacy this approach Drosophila lines containing suitable MiMIC (Minos-mediated integration cassette) transposons palette Trojan exons capable expressing range commonly used transcription factors. also exchangeable, MiMIC-like exon construct can be targeted Crispr/Cas system.

Language: Английский

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A versatile platform for creating a comprehensive UAS-ORFeome library in Drosophila

Development, Journal Year: 2013, Volume and Issue: 140(11), P. 2434 - 2442

Published: May 2, 2013

Overexpression screens are used to explore gene functions in Drosophila, but this strategy suffers from the lack of comprehensive and systematic fly strain collections efficient methods for generating such collections. Here, we present a that could be efficiently generate large numbers transgenic Drosophila strains, collection 1149 UAS-ORF lines were created with site-specific ΦC31 integrase method. For collection, set 655 genes cloned as two variants, either an open reading frame (ORF) native stop codon or C-terminal 3xHA tag. To streamline procedure generation, demonstrate utility injecting pools plasmids into embryos, each plasmid containing randomised sequence (barcode) serves unique identifier and, subsequently, strains. We also developed swapping technique facilitates rapid exchange promoters epitope tags vivo, expanding versatility ORF collection. The work described here basis library Gal4/UAS-regulated transgenes.

Language: Английский

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Optogenetic control of Drosophila using a red-shifted channelrhodopsin reveals experience-dependent influences on courtship

Nature Methods, Journal Year: 2013, Volume and Issue: 11(3), P. 325 - 332

Published: Dec. 22, 2013

Language: Английский

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A gene-specific T2A-GAL4 library for Drosophila

eLife, Journal Year: 2018, Volume and Issue: 7

Published: March 22, 2018

We generated a library of ~1000 Drosophila stocks in which we inserted construct the intron genes allowing expression GAL4 under control endogenous promoters while arresting transcription with polyadenylation signal 3' GAL4. This allows numerous applications. First, ~90% insertions essential cause severe loss-of-function phenotype, an effective way to mutagenize genes. Interestingly, 12/14 chromosomes engineered through CRISPR do not carry second-site lethal mutations. Second, 26/36 (70%) tested are rescued single UAS-cDNA construct. Third, phenotypes associated many can be reverted by excision UAS-flippase. Fourth, driven UAS-GFP/RFP reports tissue and cell-type specificity gene high sensitivity. report hundreds previously reported. Finally, cassettes replaced GFP or any DNA. These comprise powerful resource for assessing function.

Language: Английский

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