Tools and methods for cell ablation and cell inhibition in Caenorhabditis elegans - PubMed (original) (raw)

Review

Tools and methods for cell ablation and cell inhibition in Caenorhabditis elegans

Dennis Rentsch et al. Genetics. 2025.

Abstract

To understand the function of cells such as neurons within an organism, it can be instrumental to inhibit cellular function, or to remove the cell (type) from the organism, and thus to observe the consequences on organismic and/or circuit function and animal behavior. A range of approaches and tools were developed and used over the past few decades that act either constitutively or acutely and reversibly, in systemic or local fashion. These approaches make use of either drugs or genetically encoded tools. Also, there are acutely acting inhibitory tools that require an exogenous trigger like light. Here, we give an overview of such methods developed and used in the nematode Caenorhabditis elegans.

Keywords: animal rhodopsins; cell ablation; channelrhodopsins; chemical ablation; genetic ablation; hyperpolarization; laser ablation; optogenetics; photosensitizer; synaptic vesicle release.

© The Author(s) 2024. Published by Oxford University Press on behalf of The Genetics Society of America.

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Conflict of interest statement

Conflicts of interest: The authors declare no conflicts of interest.

Figures

Fig. 1.

Fig. 1.

a) Pharmacological and other silencing approaches, physical ablation, and (optogenetic) tools for inhibition of excitable cells or cell ablation, covered in this review. b) Spatial scales at which the inhibitory tools and approaches can be used. c) Temporal scales for on and off times/recovery rate of the tools and approaches. The given times are only roughly indicated for comparability. Irreversible tools are shown at the top of the chart, for simplicity.

Fig. 2.

Fig. 2.

Classes of rhodopsin-based optogenetic silencers. a) Light-driven ion pumps. b) eACRs and step function opsins for long-lasting photoinhibition. c) Natural anion channelrhodopsins. d) KCRs. Tools are color-coded according to their absorption maximum. Ions conducted or pumped, and direction based on electrochemical gradient, or pump vectoriality, are indicated.

Fig. 3.

Fig. 3.

Examples of light-sensitive GPCRs (rhodopsins), used for inhibition: a) light-evoked reversals of animals expressing MosOpn3 in ASH neurons. The multiworm tracker (Swierczek et al. 2011) was used to observe the crawling behavior of worms expressing MosOpn3 in ASH neurons. Upon blue light illumination (30 s, 100 µW/mm2, 470 nm, shaded area, 300–330 s), worms treated with ATR (100 µM) showed an increase in reversals, indicating an avoidance response upon MosOpn3 activation. b) LamPP activation in cholinergic neurons induces a wavelength-dependent increase of body length, due to muscle relaxation. Body length of animals either without ATR (−), or treated with ATR (+), were compared. Additionally, animals treated with ATR were either kept in the dark until the measurement, or preilluminated with red light (620–660 nm; overnight), or illuminated with green light (520–535 nm) for 30 s prior to the start of the experiment. Violet light illumination (5 s, 100 µW/mm2, 373–387 nm, shaded area, 5–10 s) of animals without ATR evoked no change in body length, while ATR-treated worms exhibited a slight increase. To convert the active ATR-bound state back to its inactive state, worms were illuminated with green light before the start of the measurements, resulting in a body length increase upon subsequent violet light illumination. To test whether red light illumination of the LamPP active state causes photoregeneration, worms were fed ATR under red light illumination overnight. These animals showed an increase in body length upon violet light illumination.

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