DOI: 10.5061/dryad.hqbzkh1kn
title: Drugs prescribed in Phelan-McDermid Syndrome differentially impact sensory behaviors in shank3 zebrafish models

description: Altered sensory processing is a pervasive symptom in individuals with
 Autism Spectrum Disorders (ASD). Especially in cases of profound autism,
 individuals are often medicated to manage behaviors like aggression and/or
 self-harm and/or epilepsy, and it remains unclear how these medications
 might impact perception/sensory processing. To test this, we screened
 three medications, risperidone, Lithium Chloride (LiCl), and carbamazepine
 (CBZ), prescribed to individuals with Phelan-McDermid Syndrome (PMS) and
 one drug, 2-Methyl-6-(phenylethynyl) pyridine (MPEP) tested in rodent
 models of PMS, for their effects on a sensory-induced behavior in two
 zebrafish PMS models with frameshift mutations in the N- or C- termini.
 PMS is caused by deletions of the terminal end of chromosome 22 or point
 mutations in Shank3. People with PMS can present with an array of symptoms
 including ASD, epilepsy, gastrointestinal distress, and reduced responses
 to sensory stimuli. Our zebrafish mutant shank3ab models likewise show
 reduced responses in a Visual Motor Response (VMR) assay, in which
 increased locomotion is triggered by light-to-dark transitions. To test
 how pharmacological treatments affect the VMR, we exposed larvae to
 selected drugs for twenty-four hours and then quantified their locomotion
 during four ten-minute cycles of lights on-to-off stimuli. We found that
 risperidone normalized the VMR in shank3 models, LiCl and CBZ had no
 effect on the VMR in any of the three genotypes. MPEP reduced the VMR in
 WT to levels seen in shank3 models but caused no changes in either shank3
 model. Our work shows that the effects of drugs on sensory processing is
 varied in a way that can be highly genotype- and drug-dependent.
description: <em>High-throughput behavioral screens</em>
 Behavioral experiments were performed using the DanioVision
 systemtm (Noldus, Wageningen, NTD) with the DanioVision observation
 chamber (DVOC-0040). Videos were collected at 25 fps with 1280 x 960
 resolution using a Basler acA1300-60gm camera fitted with a 12 mm
 Megapixel lens. All DanioVision experiments were run using an ANSI SBS
 compatible 96 well microtiter plate. Data were collected and analyzed
 using the DanioVision EthoVision XT  software version 11.5 (Noldus). White
 light for the visual motor response assay was set at 12% intensity on the
 high-power setting. Six-day-old larvae were acclimated to the observation
 chamber at 28°C in the dark for at least 1 hr. Visual motor response (VMR)
 experiments were run using white light cycles on-off 4 times for 5 min
 intervals for a total of 40 minutes. All behavioral experiments were
 recorded between 11 am and 3 pm, with 2-5 independent trials per
 condition. Larvae were genotyped following behavioral experiments using a
 restriction digest assay previously described (James et al., 2019; Kozol
 et al., 2021). <em>Drug
 Screening</em> Zebrafish were exposed to drugs
 dissolved in 0.1% DMSO system water (water from the system that houses the
 adult fish) 24 hours prior to running VMR assays. A range of risperidone,
 MPEP, CBZ and LiCl concentrations were derived from previously published
 papers (Tucker et al., 2006; Bruni et al., 2016; Hoffman et al., 2016),
 then dose-response curves were generated to determine an effective dose in
 relation to the VMR response of wildtype zebrafish. Concentrations used
 for comparing WT and shank3 larvae were 10 μM Risperidone (Bruni et al.,
 2016; Hoffman et al., 2016), 5 mM LiCl and 200 μM  CBZ, and 1 μM MPEP
 (Tucker et al., 2006). Genotype controls were exposed to DMSO (0.1%) in
 system water. <em>Statistics</em>
 Data were analyzed using PRISM (GraphPad, inc.) with
 Noldus-generated data. Significance was assessed using the non-parametric
 Wilcoxon rank score test (Mann-Whitney rank scores). When there were more
 than two groups, a Kruskal-Wallis rank score test was first calculated
 and, if p &lt; 0.05, was followed by a Dunn’s multiple comparisons
 test to compare all treatments and genotypes to DMSO exposed wildtype
 larvae. 
description: Programs and software used to acquire and process data;
 Daniovision behavioral box and Ethovision XT11 software, custom MATLAB
 scripts and PRISM statistical software (GraphPad inc.).