Rewiring Endogenous Bioelectric Circuits in the Xenopus laevis Embryo Model.

Area of Science:

• Developmental Biology
• Cellular Electrophysiology
• Molecular Genetics

Background:

• Embryogenesis and regeneration involve complex interactions between transcriptional and bioelectrical networks.
• Cellular resting potentials, established by ion channels and gap junctions, are crucial for organ development and tissue patterning.
• Understanding bioelectrical network roles requires methods to manipulate and monitor cellular voltage potentials during development.

Purpose of the Study:

• To describe protocols for molecular genetic targeting of ion channels and connexins using CRISPR in Xenopus laevis embryos.
• To present methods for monitoring cellular resting potential states using voltage-sensing fluorescent dye.
• To provide adaptable strategies for studying bioelectrical networks in other model organisms.

Main Methods:

• CRISPR-based molecular genetic targeting of ion channels and connexins.
• Optimization of protocols for Xenopus laevis embryos.
• Monitoring of resting potential states with voltage-sensing fluorescent dye.

Main Results:

• Established protocols for precise genetic manipulation of ion channels and connexins in developing embryos.
• Demonstrated successful monitoring of bioelectrical states using voltage-sensing fluorescent dyes.
• Validated the utility of Xenopus laevis as a tractable model for studying bioelectrical networks.

Conclusions:

• CRISPR-based genetic targeting combined with voltage-sensing dyes offers a powerful approach to investigate bioelectrical network functions in development and regeneration.
• The described protocols are adaptable to other model systems, facilitating broader research into bioelectrical regulation.
• This work advances the understanding of how bioelectrical signals contribute to fundamental biological processes.