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Automated Analysis of a Nematode Population-based Chemosensory Preference Assay

Chai, Cynthia M. and Cronin, Christopher J. and Sternberg, Paul W. (2017) Automated Analysis of a Nematode Population-based Chemosensory Preference Assay. Journal of Visualized Experiments (125). e55963. ISSN 1940-087X . PMCID PMC5612354. https://resolver.caltech.edu/CaltechAUTHORS:20170731-091632206

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Abstract

The nematode, Caenorhabditis elegans' compact nervous system of only 302 neurons underlies a diverse repertoire of behaviors. To facilitate the dissection of the neural circuits underlying these behaviors, the development of robust and reproducible behavioral assays is necessary. Previous C. elegans behavioral studies have used variations of a "drop test", a "chemotaxis assay", and a "retention assay" to investigate the response of C. elegans to soluble compounds. The method described in this article seeks to combine the complementary strengths of the three aforementioned assays. Briefly, a small circle in the middle of each assay plate is divided into four quadrants with the control and experimental solutions alternately placed. After the addition of the worms, the assay plates are loaded into a behavior chamber where microscope cameras record the worms' encounters with the treated regions. Automated video analysis is then performed and a preference index (PI) value for each video is generated. The video acquisition and automated analysis features of this method minimizes the experimenter's involvement and any associated errors. Furthermore, minute amounts of the experimental compound are used per assay and the behavior chamber's multi-camera setup increases experimental throughput. This method is particularly useful for conducting behavioral screens of genetic mutants and novel chemical compounds. However, this method is not appropriate for studying stimulus gradient navigation due to the close proximity of the control and experimental solution regions. It should also not be used when only a small population of worms is available. While suitable for assaying responses only to soluble compounds in its current form, this method can be easily modified to accommodate multimodal sensory interaction and optogenetic studies. This method can also be adapted to assay the chemosensory responses of other nematode species.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.3791/55963DOIArticle
https://www.jove.com/video/55963/automated-analysis-nematode-population-based-chemosensory-preferencePublisherArticle
https://www.jove.com/video/55963/PublisherVideo
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5612354/PubMed CentralArticle
ORCID:
AuthorORCID
Sternberg, Paul W.0000-0002-7699-0173
Additional Information:© 2017 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. Date Published: 7/13/2017. Some strains were provided by the Caenorhabditis Genetics Center (CGC), which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440). This work is supported by the Howard Hughes Medical Institute, with which P.W.S. is an investigator.
Funders:
Funding AgencyGrant Number
NIHP40 OD010440
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Subject Keywords:Behavior, Issue 125, Nematode, Caenorhabditis elegans, automated behavior analysis, population-based assay, chemosensory preference, neurobiology
Issue or Number:125
PubMed Central ID:PMC5612354
Record Number:CaltechAUTHORS:20170731-091632206
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20170731-091632206
Official Citation:Chai, C.M., Cronin, C.J., Sternberg, P.W. Automated Analysis of a Nematode Population-based Chemosensory Preference Assay. J. Vis. Exp. (125), e55963, doi:10.3791/55963 (2017).
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:79583
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:01 Aug 2017 19:48
Last Modified:03 Oct 2019 18:21

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