Minicircle-Derived scn8ab Kock-In Reporter Line Provides a Model to Study Neuronal Regulation of Zebrafish Fin Regeneration

Abstract

Understanding the cellular and molecular mechanisms of regenerative zebrafish will advance fundamental knowledge of tissue regeneration. Our previous study identified scn8ab, which encodes a voltage-gated sodium channel, as a critical gene required for fin regeneration. scn8ab likely plays roles in neurons to influence reinnervation, but its precise expression pattern during development and regeneration remained unknown. Here, I established the scn8ab knock-in (KI) reporter line (scn8abEGFP) to determine the spatiotemporal expression of scn8ab. Our minicircle technique combined with CRISPR/Cas9 gene editing enhanced the integration efficiency to create the reporter line, compared to utilizing a plasmid. Expression analysis with scn8abEGFP demonstrated neuronal expression of scn8ab in sensory and motor neurons, while non-neuronal expression was undetectable. Our scn8ab KI reporter will be essential for future studies elucidating the nerve dependence of regenerating tissues, particularly in identifying the cellular interactions between nerves and local cells in regenerating fins.