Light sheet fluorescence microscopy

Nature Reviews Methods Primers, Год журнала: 2021, Номер 1(1)

Опубликована: Ноя. 3, 2021

Язык: Английский

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Neurochemical Organization of the Drosophila Brain Visualized by Endogenously Tagged Neurotransmitter Receptors

Cell Reports, Год журнала: 2020, Номер 30(1), С. 284 - 297.e5

Опубликована: Янв. 1, 2020

Neurotransmitters often have multiple receptors that induce distinct responses in receiving cells. Expression and localization of neurotransmitter individual neurons are therefore critical for understanding the operation neural circuits. Here we describe a comprehensive library reporter strains which convertible T2A-GAL4 cassette is inserted into endogenous receptor genes Drosophila. Using this library, profile expression 75 brain. Cluster analysis reveals neurochemical segmentation brain, distinguishing higher brain centers from rest. By recombinase-mediated exchange, convert split-GFP Tango to visualize subcellular activation dopamine specific cell types. This striking differences their localization, may underlie cellular different behavioral contexts. Our resources thus provide versatile toolkit dissecting organization function systems fly

Язык: Английский

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

Journal of Experimental Biology, Год журнала: 2020, Номер 223(Suppl_1)

Опубликована: Фев. 1, 2020

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

Язык: Английский

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Cell types and neuronal circuitry underlying female aggression in Drosophila

eLife, Год журнала: 2020, Номер 9

Опубликована: Ноя. 3, 2020

Aggressive social interactions are used to compete for limited resources and regulated by complex sensory cues the organism's internal state. While both sexes exhibit aggression, its neuronal underpinnings understudied in females. Here, we identify a population of sexually dimorphic aIPg neurons adult Drosophila melanogaster central brain whose optogenetic activation increased, genetic inactivation reduced, female aggression. Analysis GAL4 lines identified an unbiased screen increased chasing behavior revealed involvement another neuron, pC1d, implicated pC1d as core nodes regulating Connectomic analysis demonstrated that interconnected suggest may exert part their effect gating flow visual information descending neurons. Our work reveals important regulatory components circuitry underlies aggressive provides tools manipulation.

Язык: Английский

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Single-cell type analysis of wing premotor circuits in the ventral nerve cord ofDrosophila melanogaster

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Июнь 1, 2023

To perform most behaviors, animals must send commands from higher-order processing centers in the brain to premotor circuits that reside ganglia distinct brain, such as mammalian spinal cord or insect ventral nerve cord. How these are functionally organized generate great diversity of animal behavior remains unclear. An important first step unraveling organization is identify their constituent cell types and create tools monitor manipulate with high specificity assess function. This possible tractable fly. a toolkit, we used combinatorial genetic technique (split-GAL4) 195 sparse driver lines targeting 198 individual These included wing haltere motoneurons, modulatory neurons, interneurons. Using combination behavioral, developmental, anatomical analyses, systematically characterized targeted our collection. Taken together, resources results presented here form powerful toolkit for future investigations neural connectivity while linking them behavioral outputs.

Язык: Английский

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An expanded toolkit for gene tagging based on MiMIC and scarless CRISPR tagging in Drosophila

eLife, Год журнала: 2018, Номер 7

Опубликована: Авг. 9, 2018

We generated two new genetic tools to efficiently tag genes in

Язык: Английский

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DrosophilaVoltage-Gated Sodium Channels Are Only Expressed in Active Neurons and Are Localized to Distal Axonal Initial Segment-like Domains

Journal of Neuroscience, Год журнала: 2020, Номер 40(42), С. 7999 - 8024

Опубликована: Сен. 14, 2020

In multipolar vertebrate neurons, action potentials (APs) initiate close to the soma, at axonal initial segment. Invertebrate neurons are typically unipolar with dendrites integrating directly into axon. Where APs initiated in axons of invertebrate is unclear. Voltage-gated sodium (Na V ) channels a functional hallmark segment vertebrates. We used an intronic Minos -Mediated Integration Cassette determine endogenous gene expression and subcellular localization sole Na channel both male female Drosophila , para . Despite being only fly, we show that 23 ± 1% embryonic larval CNS express while adult broadly expressed. generated single-cell transcriptomic atlas whole third instar brain identify expressing it positively correlates markers differentiated, actively firing neurons. Therefore, may be capable -dependent APs. then Para enriched segment, distal site dendritic integration axon, which named (DAS). The DAS present multiple neuron classes CNS. Whole cell patch clamp electrophysiological recordings fly consistent interpretation v originate DAS. Identification will enable more accurate invertebrates. SIGNIFICANCE STATEMENT potential (AP) initiation invertebrates unknown. tagged voltage-gated identified located where impinge on likely AP initiation. Understanding improves our ability model neuronal activity data. Additionally, expressed but adults. Single-cell RNA sequencing shows active, differentiated markers. able fire

Язык: Английский

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Of what use is connectomics? A personal perspective on theDrosophilaconnectome

Journal of Experimental Biology, Год журнала: 2018, Номер 221(10)

Опубликована: Май 15, 2018

ABSTRACT The brain is a network of neurons and its biological output behaviour. This an exciting age, with growing acknowledgement that the comprehensive compilation synaptic circuits densely reconstructed in brains model species now both technologically feasible scientifically enabling possibility neurobiology, much as 30 years ago genomics was molecular biology genetics. Implemented by huge advances electron microscope technology, especially focused ion beam-scanning (FIB-SEM) milling (see Glossary), image capture alignment, computer-aided reconstruction neuron morphologies, enormous progress has been made last decade detailed knowledge actual formed real neurons, various regions fly Drosophila. It useful to distinguish pathways are major, 100 or more presynaptic contacts, from those minor, fewer than about 10; most neurites postsynaptic, all sites have multiple postsynaptic dendrites. Work on Drosophila spearheaded these because cell numbers manageable, classes morphologically discrete genetically identifiable, many confirmed reporters. Recent destined within next few reveal complete connectome adult fly, paralleling larval offer same prospect possibly even shorter time frame. final amendment validation segmented bodies human proof-readers remains time-consuming step, however. value that, targeting specific transgenes either silence activate identified circuits, then identifying resulting behavioural outcome, we can determine causal mechanism for behaviour loss gain. More importantly, reveals hitherto unsuspected pathways, leading us seek novel behaviours these. Circuit information will eventually be required understand how differences between underlie behaviour, herald yet advanced connectomic strategies vertebrate brain, eventual understanding cognitive disorders having basis. Connectomes also help identify common different thus evolutionary progression candidate pathways.

Язык: Английский

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A spatially dynamic network underlies the generation of inspiratory behaviors

Proceedings of the National Academy of Sciences, Год журнала: 2019, Номер 116(15), С. 7493 - 7502

Опубликована: Март 27, 2019

Significance Breathing is a vital rhythmic behavior that originates from neural networks within the brainstem. It hypothesized breathing rhythm generated by spatially distinct localized to discrete kernels or compartments. Here, we provide evidence functional boundaries of these compartments expand and contract dynamically based on behavioral physiological demands. The ability change in size may allow be very reliable, yet flexible enough accommodate large repertoire mammalian behaviors must integrated with breathing.

Язык: Английский

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Anatomy and Neural Pathways Modulating Distinct Locomotor Behaviors in Drosophila Larva

Biology, Год журнала: 2021, Номер 10(2), С. 90 - 90

Опубликована: Янв. 25, 2021

The control of movements is a fundamental feature shared by all animals. At the most basic level, simple are generated coordinated neural activity and muscle contraction patterns that controlled central nervous system. How behavioral responses to various sensory inputs processed integrated downstream network produce flexible adaptive behaviors remains an intense area investigation in many laboratories. Due recent advances experimental techniques, pathways underlying animal have now been elucidated. For example, while role motor neurons locomotion has studied great detail, roles interneurons both noxious environments only recently realized. However, genetic transmitter identities these unclear. In this review, we provide overview circuitry required Drosophila larvae successful movements. By improving our understanding locomotor model systems such as Drosophila, will better how circuits organisms with different bodies brains lead distinct types at organism level. physiological components also provides directions understand higher organisms.

Язык: Английский

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