WormRuler: A software to track body length used to characterize a super red-shifted channelrhodopsin in Caenorhabditis elegans - PubMed (original) (raw)

WormRuler: A software to track body length used to characterize a super red-shifted channelrhodopsin in Caenorhabditis elegans

Marius Seidenthal et al. MicroPubl Biol. 2022.

Abstract

Manipulation of neuronal or muscular activity by optogenetics or other stimuli can be directly linked to the analysis of Caenorhabditis elegans ( C. elegans ) body length. Thus, WormRuler was developed as an open-source video analysis toolbox that offers video processing and data analysis in one application. Utilizing this novel tool, the super red-shifted channelrhodopsin variant, ChrimsonSA, was characterized in C. elegans . Expression and activation of ChrimsonSA in GABAergic motor neurons results in their depolarization and therefore elongation of body length, the extent of which providing information about the strength of neuronal transmission.

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Figures

Figure 1. <b> WormRuler software can be used to identify changes in body length in <i>C. elegans</i> . </b>

Figure 1.WormRuler software can be used to identify changes in body length in_C. elegans_.

**A)**The graphical user interface (GUI) of WormRuler.**B)**The input and outputs of WormRuler. (1) An example still image of a raw video recorded of a single worm. Example images of the output videos after background correction (2) and skeletonization (3). Processing steps “normalize data” (4) and “analyze data” (5) result in *.txt and *.xlsx formats, respectively.**C)**Mean (+SEM) relative body length of animals expressing_unc-47p::ChrimsonSA_ or_unc-47p::ChR2(H134R)::YFP_. Videos of animals with (+) or without (-) ATR were acquired and analyzed with WormRuler. A 10 s light stimulus (either 590-650 nm for ChrimsonSA activation or 450-490 nm for ChR2(H134R) activation) was applied after 5 s. Experiments were performed on 2-3 days with animals picked from different plates.D) Group analysis of data in (C), interval during (8-12 s) illumination with n=29 (ChrimsonSA, +ATR), n=17 (ChrimsonSA, -ATR) and n=31 (ChR2, +ATR). Unpaired t-test; *p<0.05, **p<0.01, ***p<0.001, ns not significant; error bars are s.e.m; n is the total number of animals.

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