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Drosophila optogenetics

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https://www.readbyqxmd.com/read/29757269/combining-quantitative-food-intake-assays-and-forcibly-activating-neurons-to-study-appetite-in-drosophila
#1
Lifen Jiang, Yinpeng Zhan, Yan Zhu
Food consumption is under the tight control of the brain, which integrates the physiological status, palatability, and nutritional contents of the food, and issues commands to start or stop feeding. Deciphering the processes underlying the decision-making of timely and moderate feeding carries major implications in our understanding of physiological and psychological disorders related to feeding control. Simple, quantitative, and robust methods are required to measure the food ingestion of animals after experimental manipulation, such as forcibly increasing the activities of certain target neurons...
April 24, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29681474/ejaculation-induced-by-the-activation-of-crz-neurons-is-rewarding-to-drosophila-males
#2
Shir Zer-Krispil, Hila Zak, Lisha Shao, Shir Ben-Shaanan, Lea Tordjman, Assa Bentzur, Anat Shmueli, Galit Shohat-Ophir
The reward system is a collection of circuits that reinforce behaviors necessary for survival [1, 2]. Given the importance of reproduction for survival, actions that promote successful mating induce pleasurable feeling and are positively reinforced [3, 4]. This principle is conserved in Drosophila, where successful copulation is naturally rewarding to male flies, induces long-term appetitive memories [5], increases brain levels of neuropeptide F (NPF, the fly homolog of neuropeptide Y), and prevents ethanol, known otherwise as rewarding to flies [6, 7], from being rewarding [5]...
April 17, 2018: Current Biology: CB
https://www.readbyqxmd.com/read/29630041/tracking-drosophila-larval-behavior-in-response-to-optogenetic-stimulation-of-olfactory-neurons
#3
David A Clark, Donovan Kohler, America Mathis, Eryn Slankster, Samipya Kafle, Seth R Odell, Dennis Mathew
The ability of insects to navigate toward odor sources is based on the activities of their first-order olfactory receptor neurons (ORNs). While a considerable amount of information has been generated regarding ORN responses to odorants, the role of specific ORNs in driving behavioral responses remains poorly understood. Complications in behavior analyses arise due to different volatilities of odorants that activate individual ORNs, multiple ORNs activated by single odorants, and the difficulty in replicating naturally observed temporal variations in olfactory stimuli using conventional odor-delivery methods in the laboratory...
March 21, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29618589/functional-imaging-and-optogenetics-in-drosophila
#4
Julie H Simpson, Loren L Looger
Understanding how activity patterns in specific neural circuits coordinate an animal's behavior remains a key area of neuroscience research. Genetic tools and a brain of tractable complexity make Drosophila a premier model organism for these studies. Here, we review the wealth of reagents available to map and manipulate neuronal activity with light.
April 2018: Genetics
https://www.readbyqxmd.com/read/29581265/circadian-clock-activity-of-cryptochrome-relies-on-tryptophan-mediated-photoreduction
#5
Changfan Lin, Deniz Top, Craig C Manahan, Michael W Young, Brian R Crane
Cryptochromes (CRYs) entrain the circadian clocks of plants and animals to light. Irradiation of the Drosophila cryptochrome (dCRY) causes reduction of an oxidized flavin cofactor by a chain of conserved tryptophan (Trp) residues. However, it is unclear how redox chemistry within the Trp chain couples to dCRY-mediated signaling. Here, we show that substitutions of four key Trp residues to redox-active tyrosine and redox-inactive phenylalanine tune the light sensitivity of dCRY photoreduction, conformational activation, cellular stability, and targeted degradation of the clock protein timeless (TIM)...
March 26, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29572440/a-biochemical-network-controlling-basal-myosin-oscillation
#6
Xiang Qin, Edouard Hannezo, Thomas Mangeat, Chang Liu, Pralay Majumder, Jiaying Liu, Valerie Choesmel-Cadamuro, Jocelyn A McDonald, Yiyao Liu, Bin Yi, Xiaobo Wang
The actomyosin cytoskeleton, a key stress-producing unit in epithelial cells, oscillates spontaneously in a wide variety of systems. Although much of the signal cascade regulating myosin activity has been characterized, the origin of such oscillatory behavior is still unclear. Here, we show that basal myosin II oscillation in Drosophila ovarian epithelium is not controlled by actomyosin cortical tension, but instead relies on a biochemical oscillator involving ROCK and myosin phosphatase. Key to this oscillation is a diffusive ROCK flow, linking junctional Rho1 to medial actomyosin cortex, and dynamically maintained by a self-activation loop reliant on ROCK kinase activity...
March 23, 2018: Nature Communications
https://www.readbyqxmd.com/read/29464198/unraveling-synaptic-gcamp-signals-differential-excitability-and-clearance-mechanisms-underlying-distinct-ca-2-dynamics-in-tonic-and-phasic-excitatory-and-aminergic-modulatory-motor-terminals-in-drosophila
#7
Xiaomin Xing, Chun-Fang Wu
GCaMP is an optogenetic Ca2+ sensor widely used for monitoring neuronal activities but the precise physiological implications of GCaMP signals remain to be further delineated among functionally distinct synapses. The Drosophila neuromuscular junction (NMJ), a powerful genetic system for studying synaptic function and plasticity, consists of tonic and phasic glutamatergic and modulatory aminergic motor terminals of distinct properties. We report a first simultaneous imaging and electric recording study to directly contrast the frequency characteristics of GCaMP signals of the three synapses for physiological implications...
January 2018: ENeuro
https://www.readbyqxmd.com/read/29343813/the-drosophila-gr28bd-product-is-a-non-specific-cation-channel-that-can-be-used-as-a-novel-thermogenetic-tool
#8
Aditi Mishra, Autoosa Salari, Benton R Berigan, Kayla C Miguel, Marzie Amirshenava, Abbey Robinson, Benjamin C Zars, Jenna L Lin, Lorin S Milescu, Mirela Milescu, Troy Zars
Extrinsic control of single neurons and neuronal populations is a powerful approach for understanding how neural circuits function. Adding new thermogenetic tools to existing optogenetic and other forms of intervention will increase the complexity of questions that can be addressed. A good candidate for developing new thermogenetic tools is the Drosophila gustatory receptor family, which has been implicated in high-temperature avoidance behavior. We examined the five members of the Gr28b gene cluster for temperature-dependent properties via three approaches: biophysical characterization in Xenopus oocytes, functional calcium imaging in Drosophila motor neurons, and behavioral assays in adult Drosophila...
January 17, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29284750/cyclic-amp-dependent-plasticity-underlies-rapid-changes-in-odor-coding-associated-with-reward-learning
#9
Thierry Louis, Aaron Stahl, Tamara Boto, Seth M Tomchik
Learning and memory rely on dopamine and downstream cAMP-dependent plasticity across diverse organisms. Despite the central role of cAMP signaling, it is not known how cAMP-dependent plasticity drives coherent changes in neuronal physiology that encode the memory trace, or engram. In Drosophila , the mushroom body (MB) is critically involved in olfactory classical conditioning, and cAMP signaling molecules are necessary and sufficient for normal memory in intrinsic MB neurons. To evaluate the role of cAMP-dependent plasticity in learning, we examined how cAMP manipulations and olfactory classical conditioning modulate olfactory responses in the MB with in vivo imaging...
January 16, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29259096/control-of-microtubule-dynamics-using-an-optogenetic-microtubule-plus-end-f-actin-cross-linker
#10
Rebecca C Adikes, Ryan A Hallett, Brian F Saway, Brian Kuhlman, Kevin C Slep
We developed a novel optogenetic tool, SxIP-improved light-inducible dimer (iLID), to facilitate the reversible recruitment of factors to microtubule (MT) plus ends in an end-binding protein-dependent manner using blue light. We show that SxIP-iLID can track MT plus ends and recruit tgRFP-SspB upon blue light activation. We used this system to investigate the effects of cross-linking MT plus ends and F-actin in Drosophila melanogaster S2 cells to gain insight into spectraplakin function and mechanism. We show that SxIP-iLID can be used to temporally recruit an F-actin binding domain to MT plus ends and cross-link the MT and F-actin networks...
February 5, 2018: Journal of Cell Biology
https://www.readbyqxmd.com/read/29192250/coupling-optogenetics-and-light-sheet-microscopy-a-method-to-study-wnt-signaling-during-embryogenesis
#11
Prameet Kaur, Timothy E Saunders, Nicholas S Tolwinski
Optogenetics allows precise, fast and reversible intervention in biological processes. Light-sheet microscopy allows observation of the full course of Drosophila embryonic development from egg to larva. Bringing the two approaches together allows unparalleled precision into the temporal regulation of signaling pathways and cellular processes in vivo. To develop this method, we investigated the regulation of canonical Wnt signaling during anterior-posterior patterning of the Drosophila embryonic epidermis. Cryptochrome 2 (CRY2) from Arabidopsis Thaliana was fused to mCherry fluorescent protein and Drosophila β-catenin to form an easy to visualize optogenetic switch...
November 30, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29109241/optogenetic-activation-of-the-fruitless-labeled-circuitry-in-drosophila-subobscura-males-induces-mating-motor-acts
#12
Ryoya Tanaka, Tomohiro Higuchi, Soh Kohatsu, Kosei Sato, Daisuke Yamamoto
It remains an enigma how the nervous system of different animal species produces different behaviors. We studied the neural circuitry for mating behavior in Drosophila subobscura , a species that displays unique courtship actions not shared by other members of the genera including the genetic model D. melanogaster , in which the core courtship circuitry has been identified. We disrupted the D. subobscura fruitless ( fru ) gene, a master regulator for the courtship circuitry formation in D. melanogaster , resulting in complete loss of mating behavior...
November 29, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/29097684/anion-conducting-channelrhodopsins-with-tuned-spectra-and-modified-kinetics-engineered-for-optogenetic-manipulation-of-behavior
#13
Jonas Wietek, Silvia Rodriguez-Rozada, Janine Tutas, Federico Tenedini, Christiane Grimm, Thomas G Oertner, Peter Soba, Peter Hegemann, J Simon Wiegert
Genetic engineering of natural light-gated ion channels has proven a powerful way to generate optogenetic tools for a wide variety of applications. In recent years, blue-light activated engineered anion-conducting channelrhodopsins (eACRs) have been developed, improved, and were successfully applied in vivo. We asked whether the approaches used to create eACRs can be transferred to other well-characterized cation-conducting channelrhodopsins (CCRs) to obtain eACRs with a broad spectrum of biophysical properties...
November 2, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29061981/optogenetic-neuronal-silencing-in-drosophila-during-visual-processing
#14
Alex S Mauss, Christian Busch, Alexander Borst
Optogenetic channels and ion pumps have become indispensable tools in neuroscience to manipulate neuronal activity and thus to establish synaptic connectivity and behavioral causality. Inhibitory channels are particularly advantageous to explore signal processing in neural circuits since they permit the functional removal of selected neurons on a trial-by-trial basis. However, applying these tools to study the visual system poses a considerable challenge because the illumination required for their activation usually also stimulates photoreceptors substantially, precluding the simultaneous probing of visual responses...
October 23, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28887878/simultaneous-activation-of-parallel-sensory-pathways-promotes-a-grooming-sequence-in-drosophila
#15
Stefanie Hampel, Claire E McKellar, Julie H Simpson, Andrew M Seeds
A central model that describes how behavioral sequences are produced features a neural architecture that readies different movements simultaneously, and a mechanism where prioritized suppression between the movements determines their sequential performance. We previously described a model whereby suppression drives a Drosophila grooming sequence that is induced by simultaneous activation of different sensory pathways that each elicit a distinct movement (Seeds et al., 2014). Here, we confirm this model using transgenic expression to identify and optogenetically activate sensory neurons that elicit specific grooming movements...
September 9, 2017: ELife
https://www.readbyqxmd.com/read/28887386/fragile-x-mental-retardation-protein-restricts-small-dye-iontophoresis-entry-into-central-neurons
#16
Tyler Kennedy, Kendal Broadie
Fragile X mental retardation protein (FMRP) loss causes Fragile X syndrome (FXS), a major disorder characterized by autism, intellectual disability, hyperactivity, and seizures. FMRP is both an RNA- and channel-binding regulator, with critical roles in neural circuit formation and function. However, it remains unclear how these FMRP activities relate to each other and how dysfunction in their absence underlies FXS neurological symptoms. In testing circuit level defects in the Drosophila FXS model, we discovered a completely unexpected and highly robust neuronal dye iontophoresis phenotype in the well mapped giant fiber (GF) circuit...
October 11, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28874527/dissection-of-the-drosophila-neuropeptide-f-circuit-using-a-high-throughput-two-choice-assay
#17
Lisha Shao, Mathias Saver, Phuong Chung, Qingzhong Ren, Tzumin Lee, Clement F Kent, Ulrike Heberlein
In their classic experiments, Olds and Milner showed that rats learn to lever press to receive an electric stimulus in specific brain regions. This led to the identification of mammalian reward centers. Our interest in defining the neuronal substrates of reward perception in the fruit fly Drosophila melanogaster prompted us to develop a simpler experimental approach wherein flies could implement behavior that induces self-stimulation of specific neurons in their brains. The high-throughput assay employs optogenetic activation of neurons when the fly occupies a specific area of a behavioral chamber, and the flies' preferential occupation of this area reflects their choosing to experience optogenetic stimulation...
September 19, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28835907/behavioral-and-functional-assays-for-investigating-mechanisms-of-noxious-cold-detection-and-multimodal-sensory-processing-in-drosophila-larvae
#18
Atit A Patel, Daniel N Cox
To investigate cellular, molecular and behavioral mechanisms of noxious cold detection, we developed cold plate behavioral assays and quantitative means for evaluating the predominant noxious cold-evoked contraction behavior. To characterize neural activity in response to noxious cold, we implemented a GCaMP6-based calcium imaging assay enabling in vivo studies of intracellular calcium dynamics in intact Drosophila larvae. We identified Drosophila class III multidendritic (md) sensory neurons as multimodal sensors of innocuous mechanical and noxious cold stimuli and to dissect the mechanistic bases of multimodal sensory processing we developed two independent functional assays...
July 5, 2017: Bio-protocol
https://www.readbyqxmd.com/read/28803873/neural-circuitry-that-evokes-escape-behavior-upon-activation-of-nociceptive-sensory-neurons-in-drosophila-larvae
#19
Jiro Yoshino, Rei K Morikawa, Eri Hasegawa, Kazuo Emoto
Noxious stimuli trigger a stereotyped escape response in animals. In Drosophila larvae, class IV dendrite arborization (C4 da) sensory neurons in the peripheral nervous system are responsible for perception of multiple nociceptive modalities, including noxious heat and harsh mechanical stimulation, through distinct receptors [1-9]. Silencing or ablation of C4 da neurons largely eliminates larval responses to noxious stimuli [10-12], whereas optogenetic activation of C4 da neurons is sufficient to provoke corkscrew-like rolling behavior similar to what is observed when larvae receive noxious stimuli, such as high temperature or harsh mechanical stimulation [10-12]...
August 21, 2017: Current Biology: CB
https://www.readbyqxmd.com/read/28756946/fragile-x-mental-retardation-protein-requirements-in-activity-dependent-critical-period-neural-circuit-refinement
#20
Caleb A Doll, Dominic J Vita, Kendal Broadie
Activity-dependent synaptic remodeling occurs during early-use critical periods, when naive juveniles experience sensory input. Fragile X mental retardation protein (FMRP) sculpts synaptic refinement in an activity sensor mechanism based on sensory cues, with FMRP loss causing the most common heritable autism spectrum disorder (ASD), fragile X syndrome (FXS). In the well-mapped Drosophila olfactory circuitry, projection neurons (PNs) relay peripheral sensory information to the central brain mushroom body (MB) learning/memory center...
August 7, 2017: Current Biology: CB
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