Drugs prescribed in Phelan-McDermid Syndrome differentially impact sensory behaviors in shank3 zebrafish models
Data files
Jan 02, 2023 version files 7.65 GB
-
data_directory.xlsx
11.36 KB
-
README.md
5.08 KB
-
shk3pharmacologysheets.zip
7.65 GB
Jan 05, 2023 version files 7.65 GB
-
Author_Checklist_-_Kozoletal2022.pdf
126.93 KB
-
data_directory.xlsx
11.36 KB
-
README.md
5.08 KB
-
shk3pharmacologysheets.zip
7.65 GB
Abstract
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.
Methods
High-throughput behavioral screens
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).
Drug Screening
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.
Statistics
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 < 0.05, was followed by a Dunn’s multiple comparisons test to compare all treatments and genotypes to DMSO exposed wildtype larvae.
Usage notes
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.).