Optogenetic Control of Apoptosis in Targeted Tissues of Xenopus laevis Embryos
Kyle Jewhurst1, Michael Levin1, Kelly A McLaughlin1
1Department of Biology, Center for Regenerative and Developmental Biology, Tufts University, Medford, MA, USA.
Abstract:
KillerRed (KR) is a recently discovered fluorescent protein that, when activated with green light, releases reactive oxygen species (ROS) into the cytoplasm, triggering apoptosis in a KR-expressing cell. This property allows for the use of KR as a means of killing cells in an organism with great temporal and spatial specificity, while minimizing the nonspecific effects that can result from mechanical or chemical exposure damage techniques. Such optogenetic control of cell death, and the resulting ability to induce the targeted death of specific tissues, is invaluable for regeneration/repair studies-particularly in Xenopus laevis, where apoptosis plays a key role in regeneration and repair. We here describe a method by which membrane-bound KR, introduced to Xenopus embryos by mRNA microinjection, can be activated with green light to induce apoptosis in specific organs and tissues, with a focus on the developing eye and pronephric kidney.
Insights
KillerRed (KR) protein uses light to trigger cell death (apoptosis) with precision. This optogenetic tool enables targeted tissue ablation for studying regeneration in Xenopus laevis.
Area of Science:
- Biochemistry
- Molecular Biology
- Developmental Biology
Background:
- KillerRed (KR) is a photosensitive fluorescent protein.
- KR activation releases reactive oxygen species (ROS) to induce apoptosis.
- Optogenetic control of cell death offers spatiotemporal specificity.
Purpose of the Study:
- To describe a method for using membrane-bound KR in Xenopus laevis embryos.
- To demonstrate optogenetic induction of apoptosis in specific tissues.
- To highlight applications in regeneration and repair studies.
Main Methods:
- mRNA microinjection of membrane-bound KR into Xenopus embryos.
- Activation of KR using green light.
- Analysis of apoptosis in targeted tissues, focusing on eye and kidney development.
Main Results:
- Successful induction of apoptosis in specific organs via KR activation.
- Demonstrated spatiotemporal control over cell death.
- Visualized KR-mediated apoptosis in developing eye and pronephric kidney.
Conclusions:
- Membrane-bound KR provides a precise optogenetic method for inducing apoptosis in Xenopus.
- This technique is valuable for investigating the role of apoptosis in tissue regeneration and repair.
- The study establishes a foundation for future regeneration studies using optogenetics in Xenopus.
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