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REVEALING ZEBRAFISH EMBRYONIC DEVELOPMENTAL BIOELECTRICITY USING GENETICALLY ENCODED TOOLS

thesis
posted on 2022-12-07, 03:39 authored by Martin R SilicMartin R Silic

Bioelectricity, or endogenous electrical signaling mediated by the dynamic distribution of charged molecules, is an ancient signaling mechanism conserved across living organisms. Increasing evidence has revealed that bioelectric signals play a critical role in many diverse aspects of biology such as embryonic development, cell migration, regeneration, cancer, and other diseases. However, direct visualization and manipulation of bioelectricity during development are lacking. Neuroscience has developed tools such as GEVIs (genetically encoded voltage indicators) and chemogenetics like DREADDs (designer receptor exclusively activated by designer drugs) which allow for real–time voltage monitoring and activation of mutated receptors by inert molecules for perturbing membrane potential (Vm). To uncover bioelectric activity during development, we generated a whole-zebrafish transgenic GEVI reporter line and characterized the electrical signaling during early embryogenesis using light sheet microscopy (LSM). Additionally, we generated tissue-specific transgenic lines that combined GEVIs and chemogenetic DREADD tools to manipulate Vm. We found zebrafish embryos display stage-specific characteristic bioelectric signals during the cleavage, blastula, gastrula, and segmentation periods. Furthermore, activation of DREADDs was able to alter cell-specific GEVI fluorescence intensity and could cause a melanophore hyperpigmentation phenotype. Ultimately, these results provide the first real-time systematic analysis of endogenous bioelectricity during vertebrate embryonic development. Additionally, we generated and tested zebrafish transgenic lines for simultaneous visualization and chemogenetic manipulation of Vm during development. These results provide a better understanding of developmental bioelectricity and new tools for future studies, which could eventually help uncover the cellular electric mechanisms behind tissue patterning and disease.

Funding

Roles of ion-channel mediated bioelectricity in developmental patterning

National Institute of General Medical Sciences

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History

Degree Type

  • Doctor of Philosophy

Department

  • Comparative Pathobiology

Campus location

  • West Lafayette

Advisor/Supervisor/Committee Chair

GuangJun Zhang

Additional Committee Member 2

Donna Fekete

Additional Committee Member 3

Mark Hall

Additional Committee Member 4

Timothy Ratliff

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