Abstract
Dravet syndrome (DS) is a rare developmental and epileptic encephalopathy caused by de novo mutations in the sodium channel Nav1.1 gene, SCN1A. Here, we used a translational zebrafish (ZF) model of DS (Scn1Lab) which exhibits key characteristics known to occur in patients with DS to evaluate drugs with therapeutic potential. Previous work in our laboratory has shown metabolic deficits in Scn1Lab ZF and identified 1-(2-chlorophenyl)-N-[(11)C] methyl-N-(1-methylpropyl)-3-isoquinoline carboxamide (PK11195), a mitochondrial translocator protein (TSPO) ligand, as the lead compound which decreased neuronal hyperexcitability and metabolic deficits observed in Scn1Lab mutants. In this study, we examined the effects of additional TSPO ligands, etifoxine and XBD173, on modulating behavioral and electrophysiological seizure parameters and reversing the metabolic deficits previously reported in Scn1Lab ZF. Additionally, we sought to optimize and validate a noninvasive, higher throughput multiwell multielectrode array (MEA) system to record and quantify hyperexcitability. Etifoxine and XBD173 decreased "seizure-like" swim behavior in Scn1Lab mutants. The MEA assay was validated in wild-type ZF using pentylenetetrazol and Scn1Lab mutants using PK11195 and stiripentol. The MEA assay revealed that etifoxine and XBD173 significantly inhibited neuronal hyperexcitability parameters including neuronal spikes, mean firing rate of spikes, and electrographic events. Moreover, XBD173 increased basal and maximal mitochondrial respiration. These findings suggest that TSPO may be a novel therapeutic target for treating developmental and epileptic encephalopathies such as DS. SIGNIFICANCE STATEMENT: Developmental and epileptic encephalopathies are highly drug-refractory and are in urgent need of new therapies. Current methodologies for drug discovery in larval zebrafish are limited in throughput and are highly invasive. We optimized and validated a higher throughput methodology for seizure detection in Scn1Lab mutants, identifying translocator protein, a mitochondrial protein, as a potential therapeutic target for developmental and epileptic encephalopathies.