Characterization of the zebrafish gabra1 sa43718/sa43718 germline loss of function allele confirms a function for Gabra1 in motility and nervous system development.

Autor: Reyes-Nava NG; Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX, USA., Paz D; Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX, USA., Pinales BE; Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX, USA., Perez I; Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX, USA., Gil CB; Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX, USA., Gonzales AV; Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX, USA., Grajeda BI; Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX, USA., Estevao IL; Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX, USA., Ellis CC; Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX, USA., Castro VL; Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX, USA., Quintana AM; Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX, USA. Electronic address: aquintana8@utep.edu.
Jazyk: angličtina
Zdroj: Differentiation; research in biological diversity [Differentiation] 2024 Jul-Aug; Vol. 138, pp. 100790. Date of Electronic Publication: 2024 Jun 15.
DOI: 10.1016/j.diff.2024.100790
Abstrakt: Mutation of the GABRA1 gene is associated with neurodevelopmental defects and epilepsy. GABRA1 encodes for the α1 subunit of the γ-aminobutyric acid type A receptor (GABA A R), which regulates the fast inhibitory impulses of the nervous system. Multiple model systems have been developed to understand the function of GABRA1, but these models have produced complex and, at times, incongruent data. Thus, additional model systems are required to validate and substantiate previous results. We sought to provide initial phenotypic analysis of a novel germline mutant allele. Our analysis provides a solid foundation for the future use of this allele to characterize gabra1 functionally and pharmacologically using zebrafish. We investigated the behavioral swim patterns associated with a nonsense mutation of the zebrafish gabra1 (sa43718 allele) gene. The sa43718 allele causes a decrease in gabra1 mRNA expression, which is associated with light induced hypermotility, one phenotype previously associated with seizure like behavior in zebrafish. Mutation of gabra1 was accompanied by decreased mRNA expression of gabra2, gabra3, and gabra5, indicating a reduction in the expression of additional α sub-units of the GABA A R. Although multiple sub-units were decreased, larvae continued to respond to pentylenetetrazole (PTZ), indicating that a residual GABA A R exists in the sa43718 allele. Proteomics analysis demonstrated that mutation of gabra1 is associated with abnormal expression of proteins that regulate synaptic vesicle fusion, vesicle transport, synapse development, and mitochondrial protein complexes. These data support previous studies performed in a zebrafish nonsense allele created by CRISPR/Cas9 and validate that loss of function mutations in the gabra1 gene result in seizure-like phenotypes with abnormal development of the GABA synapse. Our results add to the existing body of knowledge as to the function of GABRA1 during development and validate that zebrafish can be used to provide complete functional characterization of the gene.
(Copyright © 2024 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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