DiI staining of sensory neurons in the entomopathogenic nematode Steinernema hermaphroditum
Abstract
Steinernema hermaphroditum entomopathogenic nematodes (EPN) and their Xenorhabdus griffiniae symbiotic bacteria have recently been shown to be a genetically tractable system for the study of both parasitic and mutualistic symbiosis. In their infective juvenile (IJ) stage, EPNs search for insect hosts to invade and quickly kill them with the help of the symbiotic bacteria they contain. The mechanisms behind these behaviors have not been well characterized, including how the nematodes sense their insect hosts. In the well-studied free‑living soil nematode Caenorhabditis elegans, ciliated amphid neurons enable the worms to sense their environment, including chemosensation. Some of these neurons have also been shown to control the decision to develop as a stress-resistant dauer larva, analogous to the infective juveniles of EPNs, or to exit from dauer and resume larval development. In C. elegans and other nematodes, dye-filling with DiI is an easy and efficient method to label these neurons. We developed a protocol for DiI staining of S. hermaphroditum sensory neurons. Using this method, we could identify neurons positionally analogous to the C. elegans amphid neurons ASI, ADL, ASK, ASJ, as well as inner labial neurons IL1 and IL2. Similar to findings in other EPNs, we also found that the IJs of S. hermaphroditum are dye-filling resistant.
Additional Information
© 2022 by the authors. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. History: Received January 12, 2022 Revision received January 26, 2022 Accepted February 10, 2022 Published February 24, 2022. Funding: NSF EDGE grant 2128267 to PWS. Author Contributions: Pranjal Garg: Formal analysis, Investigation, Methodology, Visualization, Writing - original draft. Chieh-Hsiang Tan: Conceptualization, Methodology, Supervision, Writing - original draft. Paul W. Sternberg: Conceptualization, Funding acquisition, Supervision, Writing - review and editing. Reviewed By: Ray Hong.Attached Files
Published - 25789430-2022-micropub.biology.000516.pdf
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Additional details
- PMCID
- PMC8874337
- Eprint ID
- 113620
- Resolver ID
- CaltechAUTHORS:20220228-618839000
- NSF
- IOS-2128267
- Created
-
2022-02-28Created from EPrint's datestamp field
- Updated
-
2023-06-01Created from EPrint's last_modified field
- Caltech groups
- Division of Biology and Biological Engineering (BBE)