The effect of common paralytic agents used for fluorescence imaging on redox tone and ATP levels in Caenorhabditis elegans.

Autor: Morton KS; Nicholas School of Environment, Duke University, Durham, North Carolina, United States of America., Wahl AK; Nicholas School of Environment, Duke University, Durham, North Carolina, United States of America., Meyer JN; Nicholas School of Environment, Duke University, Durham, North Carolina, United States of America.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2024 Apr 26; Vol. 19 (4), pp. e0292415. Date of Electronic Publication: 2024 Apr 26 (Print Publication: 2024).
DOI: 10.1371/journal.pone.0292415
Abstrakt: One aspect of Caenorhabditis elegans that makes it a highly valuable model organism is the ease of use of in vivo genetic reporters, facilitated by its transparent cuticle and highly tractable genetics. Despite the rapid advancement of these technologies, worms must be paralyzed for most imaging applications, and few investigations have characterized the impacts of common chemical anesthetic methods on the parameters measured, in particular biochemical measurements such as cellular energetics and redox tone. Using two dynamic reporters, QUEEN-2m for relative ATP levels and reduction-oxidation sensitive GFP (roGFP) for redox tone, we assess the impact of commonly used chemical paralytics. We report that no chemical anesthetic is entirely effective at doses required for full paralysis without altering redox tone or ATP levels, and that anesthetic use alters the detected outcome of rotenone exposure on relative ATP levels and redox tone. We also assess the use of cold shock, commonly used in combination with physical restraint methods, and find that cold shock does not alter either ATP levels or redox tone. In addition to informing which paralytics are most appropriate for research in these topics, we highlight the need for tailoring the use of anesthetics to different endpoints and experimental questions. Further, we reinforce the need for developing less disruptive paralytic methods for optimal imaging of dynamic in vivo reporters.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2024 Morton et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
Databáze: MEDLINE
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