Abstract
Genetic engineering techniques have contributed to the now widespread use of zebrafish to investigate gene function, but zebrafish-based human disease studies, and particularly for neurological disorders, are limited. Here we used CRISPR-Cas9 to generate 40 single-gene mutant zebrafish lines representing catastrophic childhood epilepsies. We evaluated larval phenotypes using electrophysiological, behavioral, neuro-anatomical, survival and pharmacological assays. Local field potential recordings (LFP) were used to screen ∼3300 larvae. Phenotypes with unprovoked electrographic seizure activity (i.e., epilepsy) were identified in zebrafish lines for 8 genes; ARX, EEF1A, GABRB3, GRIN1, PNPO, SCN1A, STRADA and STXBP1. We also created an open-source database containing sequencing information, survival curves, behavioral profiles and representative electrophysiology data. We offer all zebrafish lines as a resource to the neuroscience community and envision them as a starting point for further functional analysis and/or identification of new therapies.
Highlights
Genetic engineering techniques have contributed to the widespread use of zebrafish to investigate gene function, but zebrafish-based human disease studies, and for neurological disorders, are limited
Seizure classifications promoted by the International League Against Epilepsy (ILAE)[33] are largely defined by the presence of unprovoked “self-sustained paroxysmal disorders of brain function” and these classification resources have evolved continuously since the 1960s
We evaluated genes identified in humans using genomewide association (GWAS) and pathological de novo variants from trio exome sequencing studies[6,7,8,9,10,41]
Summary
Genetic engineering techniques have contributed to the widespread use of zebrafish to investigate gene function, but zebrafish-based human disease studies, and for neurological disorders, are limited. Seizure classifications promoted by the International League Against Epilepsy (ILAE)[33] are largely defined by the presence of unprovoked “self-sustained paroxysmal disorders of brain function” and these classification resources have evolved continuously since the 1960s Adaptation of this strategy to animal models[34], zebrafish models developed for catastrophic epilepsies of childhood, is difficult. We established readouts to identify seizures at electrographic and behavioral levels, and an opensource online website to efficiently share data with the neuroscience community As many of these zebrafish represent rare genetic diseases for which our understanding of pathophysiology remains largely unknown, they provide a rich resource to further investigate key etiological questions or utilization in highthroughput precision medicine-based therapy development
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
