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Drosophila optogenetics

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https://www.readbyqxmd.com/read/27818173/the-trp-channels-pkd2-nompc-and-trpm-act-in-cold-sensing-neurons-to-mediate-unique-aversive-behaviors-to-noxious-cold-in-drosophila
#1
Heather N Turner, Kevin Armengol, Atit A Patel, Nathaniel J Himmel, Luis Sullivan, Srividya Chandramouli Iyer, Surajit Bhattacharya, Eswar Prasad R Iyer, Christian Landry, Michael J Galko, Daniel N Cox
The basic mechanisms underlying noxious cold perception are not well understood. We developed Drosophila assays for noxious cold responses. Larvae respond to near-freezing temperatures via a mutually exclusive set of singular behaviors-in particular, a full-body contraction (CT). Class III (CIII) multidendritic sensory neurons are specifically activated by cold and optogenetic activation of these neurons elicits CT. Blocking synaptic transmission in CIII neurons inhibits CT. Genetically, the transient receptor potential (TRP) channels Trpm, NompC, and Polycystic kidney disease 2 (Pkd2) are expressed in CIII neurons, where each is required for CT...
December 5, 2016: Current Biology: CB
https://www.readbyqxmd.com/read/27730580/performing-chromophore-assisted-laser-inactivation-in-drosophila-embryos-using-gfp
#2
Anne Pélissier-Monier, Bénédicte Sanson, Bruno Monier
Chromophore-assisted laser inactivation (CALI) is an optogenetic technique in which light-induced release of reactive oxygen species triggers acute inactivation of a protein of interest, with high spatial and temporal resolution. At its simplest, selective protein inactivation can be achieved via the genetic fusion of the protein to a photosensitizer such as EGFP, and using standard optical setups such as laser scanning confocal microscopes. Although use of CALI in Drosophila is relatively recent, this technique can be a powerful complement to developmental genetics, especially in vivo as it allows visualization of the immediate consequences of local protein inactivation when coupled to time-lapse microscopy analysis...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27617442/odorant-responses-and-courtship-behaviors-influenced-by-at4-neurons-in-drosophila
#3
Svetlana Pitts, Elizabeth Pelser, Julian Meeks, Dean Smith
In insects, pheromones function as triggers to elicit complex behavior programs, such as courtship and mating behavior. In most species, the neurons tuned to pheromones are localized in a specific subset of olfactory sensilla located on the antenna called trichoid sensilla. In Drosophila there are two classes of trichoid sensilla, at1 sensilla that contain the dendrites of a single neuron that is specifically tuned to the male-specific pheromone 11-cis vaccenyl acetate (cVA), and at4 sensilla that contain three neurons with relatively poorly defined chemical specificity and function...
2016: PloS One
https://www.readbyqxmd.com/read/27585032/olfactory-behaviors-assayed-by-computer-tracking-of-drosophila-in-a-four-quadrant-olfactometer
#4
Chun-Chieh Lin, Olena Riabinina, Christopher J Potter
A key challenge in neurobiology is to understand how neural circuits function to guide appropriate animal behaviors. Drosophila melanogaster is an excellent model system for such investigations due to its complex behaviors, powerful genetic techniques, and compact nervous system. Laboratory behavioral assays have long been used with Drosophila to simulate properties of the natural environment and study the neural mechanisms underlying the corresponding behaviors (e.g. phototaxis, chemotaxis, sensory learning and memory)(1-3)...
2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27572257/functional-integration-of-a-serotonergic-neuron-in-the-drosophila-antennal-lobe
#5
Xiaonan Zhang, Quentin Gaudry
Serotonin plays a critical role in regulating many behaviors that rely on olfaction and recently there has been great effort in determining how this molecule functions in vivo. However, it remains unknown how serotonergic neurons that innervate the first olfactory relay respond to odor stimulation and how they integrate synaptically into local circuits. We examined the sole pair of serotonergic neurons that innervates the Drosophila antennal lobe (the first olfactory relay) to characterize their physiology, connectivity, and contribution to pheromone processing...
2016: ELife
https://www.readbyqxmd.com/read/27484401/high-brightness-organic-light-emitting-diodes-for-optogenetic-control-of-drosophila-locomotor-behaviour
#6
Andrew Morton, Caroline Murawski, Stefan R Pulver, Malte C Gather
Organic light emitting diodes (OLEDs) are in widespread use in today's mobile phones and are likely to drive the next generation of large area displays and solid-state lighting. Here we show steps towards their utility as a platform technology for biophotonics, by demonstrating devices capable of optically controlling behaviour in live animals. Using devices with a pin OLED architecture, sufficient illumination intensity (0.3 mW.mm(-2)) to activate channelrhodopsins (ChRs) in vivo was reliably achieved at low operating voltages (5 V)...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27479324/circadian-neuron-feedback-controls-the-drosophila-sleep-activity-profile
#7
Fang Guo, Junwei Yu, Hyung Jae Jung, Katharine C Abruzzi, Weifei Luo, Leslie C Griffith, Michael Rosbash
Little is known about the ability of Drosophila circadian neurons to promote sleep. Here we show, using optogenetic manipulation and video recording, that a subset of dorsal clock neurons (DN1s) are potent sleep-promoting cells that release glutamate to directly inhibit key pacemaker neurons. The pacemakers promote morning arousal by activating these DN1s, implying that a late-day feedback circuit drives midday siesta and night-time sleep. To investigate more plastic aspects of the sleep program, we used a calcium assay to monitor and compare the real-time activity of DN1 neurons in freely behaving males and females...
August 18, 2016: Nature
https://www.readbyqxmd.com/read/27445732/artificial-induction-of-associative-olfactory-memory-by-optogenetic-and-thermogenetic-activation-of-olfactory-sensory-neurons-and-octopaminergic-neurons-in-drosophila-larvae
#8
Takato Honda, Chi-Yu Lee, Ken Honjo, Katsuo Furukubo-Tokunaga
The larval brain of Drosophila melanogaster provides an excellent system for the study of the neurocircuitry mechanism of memory. Recent development of neurogenetic techniques in fruit flies enables manipulations of neuronal activities in freely behaving animals. This protocol describes detailed steps for artificial induction of olfactory associative memory in Drosophila larvae. In this protocol, the natural reward signal is substituted by thermogenetic activation of octopaminergic neurons in the brain. In parallel, the odor signal is substituted by optogenetic activation of a specific class of olfactory receptor neurons...
2016: Frontiers in Behavioral Neuroscience
https://www.readbyqxmd.com/read/27441388/dopaminergic-neurons-write-and-update-memories-with-cell-type-specific-rules
#9
Yoshinori Aso, Gerald M Rubin
Associative learning is thought to involve parallel and distributed mechanisms of memory formation and storage. In Drosophila, the mushroom body (MB) is the major site of associative odor memory formation. Previously we described the anatomy of the adult MB and defined 20 types of dopaminergic neurons (DANs) that each innervate distinct MB compartments (Aso et al., 2014a, 2014b). Here we compare the properties of memories formed by optogenetic activation of individual DAN cell types. We found extensive differences in training requirements for memory formation, decay dynamics, storage capacity and flexibility to learn new associations...
2016: ELife
https://www.readbyqxmd.com/read/27373835/behavior-reveals-selective-summation-and-max-pooling-among-olfactory-processing-channels
#10
Joseph S Bell, Rachel I Wilson
The olfactory system is divided into processing channels (glomeruli), each receiving input from a different type of olfactory receptor neuron (ORN). Here we investigated how glomeruli combine to control behavior in freely walking Drosophila. We found that optogenetically activating single ORN types typically produced attraction, although some ORN types produced repulsion. Attraction consisted largely of a behavioral program with the following rules: at fictive odor onset, flies walked upwind, and at fictive odor offset, they reversed...
July 20, 2016: Neuron
https://www.readbyqxmd.com/read/27281494/cyclic-nucleotide-signaling-is-required-during-synaptic-refinement-at-the-drosophila-neuromuscular-junction
#11
Fernando Vonhoff, Haig Keshishian
The removal of miswired synapses is a fundamental prerequisite for normal circuit development, leading to clinical problems when aberrant. However, the underlying activity-dependent molecular mechanisms involved in synaptic pruning remain incompletely resolved. Here we examine the dynamic properties of intracellular calcium oscillations and test a role for cAMP signaling during synaptic refinement in intact Drosophila embryos using optogenetic tools. We provide in vivo evidence at the single gene level that the calcium-dependent adenylyl cyclase rutabaga, the phosphodiesterase dunce, the kinase PKA, and Protein Phosphatase 1 (PP1) all operate within a functional signaling pathway to modulate Sema2a-dependent chemorepulsion...
June 9, 2016: Developmental Neurobiology
https://www.readbyqxmd.com/read/27269519/the-timed-depolarization-of-morning-and-evening-oscillators-phase-shifts-the-circadian-clock-of-drosophila
#12
Saskia Eck, Charlotte Helfrich-Förster, Dirk Rieger
Phase response curves (PRCs) for light or temperature stimuli have been shown to be most valuable in understanding how circadian clocks are entrained to daily environmental cycles. Nowadays, PRC experiments in which clock neurons are manipulated in a temporally restricted manner by thermogenetic or optogenetic tools are also useful to comprehend clock network properties. Here, we temporally depolarized specific clock neurons of Drosophila melanogaster by activating temperature-sensitive dTrpA1 channels to unravel their role in phase shifting the flies' activity rhythm...
October 2016: Journal of Biological Rhythms
https://www.readbyqxmd.com/read/27214889/fim2c-a-multi-colour-multi-purpose-imaging-system-to-manipulate-and-analyse-animal-behaviour
#13
Benjamin Risse, Nils Otto, Dimitri Berh, Xiaoyi Jiang, Matthias Kiel, Christian Klambt
In vivo whole-body imaging of small animals plays an important role for biomedical studies. In particular, animals like the fruit fly Drosophila melanogaster or the nematode Caenorhabditis elegans are popular model organisms for preclinical research since they offer sophisticated genetic tool-kits. Recording these translucent animals with high contrast in a large arena is however not trivial. Furthermore fluorescent proteins are widely used to mark cells in vivo and report their functions. This paper introduces a novel optical imaging technique called FIM2c enabling simultaneous detection of the animals posture and movement as well as fluorescent markers like GFP...
May 18, 2016: IEEE Transactions on Bio-medical Engineering
https://www.readbyqxmd.com/read/27077482/high-throughput-assay-to-examine-egg-laying-preferences-of-individual-drosophila-melanogaster
#14
Bin Gou, Edward Zhu, Ruo He, Ulrich Stern, Chung-Hui Yang
Recently, egg-laying preference of Drosophila has emerged as a genetically tractable model to study the neural basis of simple decision-making processes. When selecting sites to deposit their eggs, female flies are capable of ranking the relative attractiveness of their options and choosing the "greater of two goods." However, most egg-laying preference assays are not practical if one wants to take a systematic genetic screening approach to search for the circuit basis underlying this simple decision-making process, as they are population-based and laborious to set up...
2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27073064/astrocytic-glutamate-transport-regulates-a-drosophila-cns-synapse-that-lacks-astrocyte-ensheathment
#15
Sarah E MacNamee, Kendra E Liu, Stephan Gerhard, Cathy T Tran, Richard D Fetter, Albert Cardona, Leslie P Tolbert, Lynne A Oland
Anatomical, molecular, and physiological interactions between astrocytes and neuronal synapses regulate information processing in the brain. The fruit fly Drosophila melanogaster has become a valuable experimental system for genetic manipulation of the nervous system and has enormous potential for elucidating mechanisms that mediate neuron-glia interactions. Here, we show the first electrophysiological recordings from Drosophila astrocytes and characterize their spatial and physiological relationship with particular synapses...
July 1, 2016: Journal of Comparative Neurology
https://www.readbyqxmd.com/read/27011354/sensitive-red-protein-calcium-indicators-for-imaging-neural-activity
#16
Hod Dana, Boaz Mohar, Yi Sun, Sujatha Narayan, Andrew Gordus, Jeremy P Hasseman, Getahun Tsegaye, Graham T Holt, Amy Hu, Deepika Walpita, Ronak Patel, John J Macklin, Cornelia I Bargmann, Misha B Ahrens, Eric R Schreiter, Vivek Jayaraman, Loren L Looger, Karel Svoboda, Douglas S Kim
Genetically encoded calcium indicators (GECIs) allow measurement of activity in large populations of neurons and in small neuronal compartments, over times of milliseconds to months. Although GFP-based GECIs are widely used for in vivo neurophysiology, GECIs with red-shifted excitation and emission spectra have advantages for in vivo imaging because of reduced scattering and absorption in tissue, and a consequent reduction in phototoxicity. However, current red GECIs are inferior to the state-of-the-art GFP-based GCaMP6 indicators for detecting and quantifying neural activity...
2016: ELife
https://www.readbyqxmd.com/read/26965137/optogenetic-manipulation-of-selective-neural-activity-in-free-moving-drosophila-adults
#17
Po-Yen Hsiao, Ming-Chin Wu, Yen-Yin Lin, Chein-Chung Fu, Ann-Shyn Chiang
Activating selected neurons elicits specific behaviors in Drosophila adults. By combining optogenetics and laser-tracking techniques, we have recently developed an automated laser-tracking and optogenetic manipulation system (ALTOMS) for studying how brain circuits orchestrate complex behaviors. The established ALTOMS can independently target three lasers (473-nm blue laser, 593.5-nm yellow laser, and 1064-nm infrared laser) on any specified body part of two freely moving flies. Triggering light-sensitive proteins in real time, the blue laser and yellow laser can respectively activate and inhibit target neurons in artificial transgenic flies...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/26965122/optogenetics-in-drosophila-neuroscience
#18
Thomas Riemensperger, Robert J Kittel, André Fiala
Optogenetic techniques enable one to target specific neurons with light-sensitive proteins, e.g., ion channels, ion pumps, or enzymes, and to manipulate their physiological state through illumination. Such artificial interference with selected elements of complex neuronal circuits can help to determine causal relationships between neuronal activity and the effect on the functioning of neuronal circuits controlling animal behavior. The advantages of optogenetics can best be exploited in genetically tractable animals whose nervous systems are, on the one hand, small enough in terms of cell numbers and to a certain degree stereotypically organized, such that distinct and identifiable neurons can be targeted reproducibly...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/26919434/parallel-transformation-of-tactile-signals-in-central-circuits-of-drosophila
#19
John C Tuthill, Rachel I Wilson
To distinguish between complex somatosensory stimuli, central circuits must combine signals from multiple peripheral mechanoreceptor types, as well as mechanoreceptors at different sites in the body. Here, we investigate the first stages of somatosensory integration in Drosophila using in vivo recordings from genetically labeled central neurons in combination with mechanical and optogenetic stimulation of specific mechanoreceptor types. We identify three classes of central neurons that process touch: one compares touch signals on different parts of the same limb, one compares touch signals on right and left limbs, and the third compares touch and proprioceptive signals...
February 25, 2016: Cell
https://www.readbyqxmd.com/read/26877086/four-individually-identified-paired-dopamine-neurons-signal-reward-in-larval-drosophila
#20
Astrid Rohwedder, Nana L Wenz, Bernhard Stehle, Annina Huser, Nobuhiro Yamagata, Marta Zlatic, James W Truman, Hiromu Tanimoto, Timo Saumweber, Bertram Gerber, Andreas S Thum
Dopaminergic neurons serve multiple functions, including reinforcement processing during associative learning [1-12]. It is thus warranted to understand which dopaminergic neurons mediate which function. We study larval Drosophila, in which only approximately 120 of a total of 10,000 neurons are dopaminergic, as judged by the expression of tyrosine hydroxylase (TH), the rate-limiting enzyme of dopamine biosynthesis [5, 13]. Dopaminergic neurons mediating reinforcement in insect olfactory learning target the mushroom bodies, a higher-order "cortical" brain region [1-5, 11, 12, 14, 15]...
March 7, 2016: Current Biology: CB
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