Dissecting a circuit for olfactory behaviour in Caenorhabditis elegans

Chalasani, Sreekanth H.; Chronis, Nikos; Tsunozaki, Makoto; Gray, Jesse; Ramot, Daniel; Goodman, Miriam B.; Bargmann, Cornelia I.

doi:10.1038/nature06292

Cited by 593 publications

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“…This method enables sensitive measurements of action potential patterns and shows whether responses are excitatory or inhibitory, but drawing final conclusions about the individual firing neuron is difficult, as each sensillum contains two to four neurons 14 . In the larva, multicellular electrophysiological measurements of the taste system have been performed, but this method is technically challenging and it is not clear how many taste neurons are housed in which sensillum 15 .Microfluidic devices have been used previously to image command interneurons (AVA) and sensory neurons (ASH) in the nematode Caenorhabditis elegans and for studies of axotomy in peripheral sensory neurons of Drosophila larvae [16][17][18] . To implement a similar strategy for Drosophila larval chemosensation, we developed a simple and widely applicable microfluidic chip.…”

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confidence: 99%

“…Microfluidic devices have been used previously to image command interneurons (AVA) and sensory neurons (ASH) in the nematode Caenorhabditis elegans and for studies of axotomy in peripheral sensory neurons of Drosophila larvae [16][17][18] . To implement a similar strategy for Drosophila larval chemosensation, we developed a simple and widely applicable microfluidic chip.…”

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A microfluidics-based method for measuring neuronal activity in Drosophila chemosensory neurons

et al. 2016

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Animals possess highly specialized sensory neurons that are capable of perceiving information about a continuously changing environment. Measuring neuronal activity in these cells is an important step toward understanding the basic coding principles, as well as the spatial and temporal dynamics, of sensory systems. The fruit fly Drosophila melanogaster is widely used as a genetic model organism to uncover basic mechanisms of taste coding and odor perception [1][2][3][4][5][6][7] . Recent insights into the molecular and functional mechanisms of sensory perception in Drosophila have been achieved by the use of single-sensillum recordings of adult taste or olfactory sensilla 1,4,[8][9][10][11][12][13] . This method enables sensitive measurements of action potential patterns and shows whether responses are excitatory or inhibitory, but drawing final conclusions about the individual firing neuron is difficult, as each sensillum contains two to four neurons 14 . In the larva, multicellular electrophysiological measurements of the taste system have been performed, but this method is technically challenging and it is not clear how many taste neurons are housed in which sensillum 15 .Microfluidic devices have been used previously to image command interneurons (AVA) and sensory neurons (ASH) in the nematode Caenorhabditis elegans and for studies of axotomy in peripheral sensory neurons of Drosophila larvae [16][17][18] . To implement a similar strategy for Drosophila larval chemosensation, we developed a simple and widely applicable microfluidic chip. We used this custom-made microfluidic device to deliver precise and temporally restricted tastant stimulation in the first published physiological characterization of individual larval gustatory receptor neurons (GRNs) expressing the calcium sensor GCaMP5 (ref. 19). Development of the protocolThe design of the chip is simple. It contains a single channel that is connected to a chamber in which the sample can be mounted to bring it into contact with fluids in the channel. The chamber was originally designed to exactly fit the head of a third-instar Drosophila larva; however, it is also suitable for the proboscis or the leg of an adult fly. The channel for delivering stimulants is relatively broad, designed at a width of 1 mm to reduce flow pressure on the sample. We use a single-channel system in which changes of liquids are performed by the tubing system outside the chip, allowing a relatively slow flow rate inside the chip. Here we provide information required for the design and production of this microfluidic device ( Fig. 1; Supplementary Data), as well as a detailed protocol for live imaging to acquire GCaMP-based calcium imaging (Figs. 2 and 3). By introducing the larval head, or the adult proboscis or leg, into the chamber, neuronal activity can be recorded while chemosensory neurons are in contact with the solution pumped through the channel (Figs. 2 and 3).Overview of the procedure An overview of the workflow involved in implementing the procedure is provided in Figu...

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A microfluidics-based method for measuring neuronal activity in Drosophila chemosensory neurons

et al. 2016

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“…17,18 As noted above, sensory information about food availability is also transmitted through mechanosensory activation of dopaminergic neurons (CEP, ADE and PDE). 19 What are the respective contributions of these sensory modalities to local food searching?…”

Section: The Interneuron Dva and Local Search Behaviormentioning

confidence: 99%

In the proper context: Neuropeptide regulation of behavioral transitions during food searching

Bhattacharya

Francis

2015

Worm

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“…8 8 Flash and freeze electron microscopy for the time-dependent analysis of the synaptic ultrastructure following optogenetic stimulation. a Scheme of the experiment for slow synaptic processes, induced by extreme activity.…”

Section: Optogenetic Analysis Of Pain Receptor Genesmentioning

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Optogenetic analyses of neuronal network function and synaptic transmission in Caenorhabditis elegans

Gottschalk

2014

E-Neuroforum

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Caenorhabditis elegans is a popular model organism in the neurosciences [4]. This animal stands out in particular due to a (seemingly) simple nervous system consisting of only 302 neurons. Based on the fundamental research by John White and colleagues from the 1970s and 1980s, these neurons are precisely mapped in the entirety of their synaptic connections (chemical and electrical synapses; [51]): by means of serial electron microscopy, the entire nervous system, synapse by synapse and including all anatomical connections, has been unraveled. Since eutely (having a fixed number of somatic cells) occurs in every single individual of C. elegans and since it can be expected that the synapses are genetically determined and not destined by plasticity or learning, one can expect consistency of synapses between individuals. However, evidence for this still needs to be provided by serial electron microscopy and connectomics being conducted on several individuals; even with modern block-face imaging and semi-automatic segmentation, this requires huge effort. White et al. [51] have been able to map about 7000 synapses in between the 302 neurons and muscle cells. In this neuronal network, elementary circuits can be found which, in an analogous, logical way of operation, can also be found in the nervous system of higher animals, with the difference that they exist in myriads of copies and work in parallel in order to comply with more complex tasks. The majority of synaptic key factors is highly conserved between humans and nematodes and, in most cases, these have been identified in C. elegans for the first time [6,29]. Thus, the nervous system of C. elegans is not only a valid model for higher nervous systems on the protein level, but also on the level of basic circuits or logical circuitry modules. Simple nervous system, complex behaviorAs simple as C. elegans seems to be as an animal, the more versatile upon closer analysis are the behavioral patterns this animal is able to generate [39]. C. elegans carries out several types of locomotion, navigation in two and three dimensions, as well as complex mating behaviors [5]. The nematode has sensors for numerous modalities, e.g., temperature, chemicals, tastants, oxygen and CO 2 , perception of light and magnetic fields, as well as various types of mechano-and nociception. Most of these sensory modalities lead to responses in the form of taxis, escape behavior, or altered navigation. However, long-term changes in behavior can also occur, as well as switching between different behavioral states (mainly regulated by neuromodulators). The animal exhibits several types of quiescence behaviors, which feature characteristics of sleep [31]. Furthermore, the nervous system features habituation as well as simple forms of (associative) learning [2,34]. Single modes of behavior are often controlled by a few neurons that form simple circuits. This means that the control of behavior is determined by the physiological characteristics and synaptic connections of the 302 neurons.Cultivation of...

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Dissecting a circuit for olfactory behaviour in Caenorhabditis elegans

Cited by 593 publications

References 50 publications

A microfluidics-based method for measuring neuronal activity in Drosophila chemosensory neurons

A microfluidics-based method for measuring neuronal activity in Drosophila chemosensory neurons

In the proper context: Neuropeptide regulation of behavioral transitions during food searching

Optogenetic analyses of neuronal network function and synaptic transmission in Caenorhabditis elegans

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