Automated imaging of neuronal activity in freely behaving Caenorhabditis elegans

Arous, Juliette Ben; Tanizawa, Yoshinori; Rabinowitch, Ithai; Chatenay, D.; Schafer, William R.

doi:10.1016/j.jneumeth.2010.01.011

Cited by 88 publications

(83 citation statements)

References 26 publications

(29 reference statements)

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“…S5 A and B). Because calcium influx of AVA is correlated with the initiation of spontaneous backward movements (26,35,36), we recorded AVA calcium dynamics of animals in the nematode growth medium (NGM) buffer using a microfluidic device that allows generation of body waves, which result in forward or backward movements (36). We measured the G-CaMP fluorescence intensity in AVA in naive and PA14-trained animals and calculated the rate of intensity change (Fig.…”

Section: Dbl-1 Signals To the Hypodermis To Direct Aversive Olfactorymentioning

confidence: 99%

DBL-1, a TGF-β, is essential for Caenorhabditis elegans aversive olfactory learning

Zhang¹,

Zhang

2012

Proc. Natl. Acad. Sci. U.S.A.

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The TGF-β superfamily is conserved throughout metazoan, and its members play essential roles in development and disease. TGF-β has also been implicated in adult neural plasticity. However, the underlying mechanisms are not well understood. Here we report that DBL-1, a Caenorhabditis elegans TGF-β homolog known to control body morphology and immunity, is essential for aversive olfactory learning of potentially harmful bacteria food. We show that DBL-1 generated by the AVA command interneurons, which are critical for sensorimotor responses, regulates aversive olfactory learning, and that the activity of the type I TGF-β receptor SMA-6 in the hypodermis is needed during adulthood to generate olfactory plasticity. These spatial and temporal mechanisms are critical for the DBL-1 signaling to achieve its diverse functions in development and adult neural plasticity. Interestingly, aversive training decreases AVA calcium response, leading to an increase in the DBL-1 signal secreted from AVA, revealing an experience-dependent change that can underlie the role of TGF-β signaling in mediating plasticity.experience-dependent secretion | molecular and cellular mechanisms for TGF-beta pathway in learning

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Section: Dbl-1 Signals To the Hypodermis To Direct Aversive Olfactorymentioning

confidence: 99%

DBL-1, a TGF-β, is essential for Caenorhabditis elegans aversive olfactory learning

Zhang¹,

Zhang

2012

Proc. Natl. Acad. Sci. U.S.A.

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“…15. To facilitate higher-throughput recordings of behavior, a number of tracking microscopes (16)(17)(18) or multiworm imagers were developed (19,20) along with sophisticated behavioral analysis software and analytical tools (21,22). Motivated in part to understand these behaviors, calcium imaging systems were Significance Large-scale neural recordings in freely moving animals are important for understanding how patterns of activity across a population of neurons generates animal behavior.…”

mentioning

confidence: 99%

Whole-brain calcium imaging with cellular resolution in freely behaving Caenorhabditis elegans

Nguyen

Shipley

Linder

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

345

355

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The ability to acquire large-scale recordings of neuronal activity in awake and unrestrained animals is needed to provide new insights into how populations of neurons generate animal behavior. We present an instrument capable of recording intracellular calcium transients from the majority of neurons in the head of a freely behaving Caenorhabditis elegans with cellular resolution while simultaneously recording the animal's position, posture, and locomotion. This instrument provides whole-brain imaging with cellular resolution in an unrestrained and behaving animal. We use spinning-disk confocal microscopy to capture 3D volumetric fluorescent images of neurons expressing the calcium indicator GCaMP6s at 6 head-volumes/s. A suite of three cameras monitor neuronal fluorescence and the animal's position and orientation. Custom software tracks the 3D position of the animal's head in real time and two feedback loops adjust a motorized stage and objective to keep the animal's head within the field of view as the animal roams freely. We observe calcium transients from up to 77 neurons for over 4 min and correlate this activity with the animal's behavior. We characterize noise in the system due to animal motion and show that, across worms, multiple neurons show significant correlations with modes of behavior corresponding to forward, backward, and turning locomotion.calcium imaging | large-scale recording | behavior | C. elegans | microscopy H ow do patterns of neural activity generate an animal's behavior? To answer this question, it is important to develop new methods for recording from large populations of neurons in animals as they move and behave freely. The collective activity of many individual neurons appears to be critical for generating behaviors including arm reach in primates (1), song production in zebrafinch (2), the choice between swimming or crawling in leech (3), and decision-making in mice during navigation (4). New methods for recording from larger populations of neurons in unrestrained animals are needed to better understand neural coding of these behaviors and neural control of behavior more generally.Calcium imaging has emerged as a promising technique for recording dynamics from populations of neurons. Calcium-sensitive proteins are used to visualize changes in intracellular calcium levels in neurons in vivo which serve as a proxy for neural activity (5). To resolve the often weak fluorescent signal of an individual neuron in a dense forest of other labeled cells requires a high magnification objective with a large numerical aperture that, consequently, can image only a small field of view. Calcium imaging has traditionally been performed on animals that are stationary from anesthetization or immobilization to avoid imaging artifacts induced by animal motion. As a result, calcium imaging studies have historically focused on small brain regions in immobile animals that exhibit little or no behavior (6).No previous neurophysiological study has attained whole-brain imaging with cellular resolution in a...

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“…Since differences in the susceptibility of the donor-and acceptor-FPs to photobleaching could affect ratiometric measurements, it is particularly important to use FPs that are photostable and free from photo-switching behavior. elegans, Drosophila, and Zebrafish have been generated that express calcium sensing biosensor proteins [9][10][11] . In general, the imaging of biosensor activities in transgenic mice, however, has proven to be more difficult 12 .…”

Section: ) Optimized Probes For Tplsmmentioning

confidence: 99%

A simple approach for measuring FRET in fluorescent biosensors using two-photon microscopy

2016

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Automated imaging of neuronal activity in freely behaving Caenorhabditis elegans

Cited by 88 publications

References 26 publications

DBL-1, a TGF-β, is essential for Caenorhabditis elegans aversive olfactory learning

DBL-1, a TGF-β, is essential for Caenorhabditis elegans aversive olfactory learning

Whole-brain calcium imaging with cellular resolution in freely behaving Caenorhabditis elegans

A simple approach for measuring FRET in fluorescent biosensors using two-photon microscopy

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