DOI: 10.5061/dryad.cnp5hqc2m
title: Vape flavorants dull sensory perception and cause hyperactivity in developing zebrafish embryos

description: E-cigarette use (vaping) during pregnancy has been increasing, and the
 potential exists for the developing brain in utero to be exposed to
 chemical constituents in the vape. Vapes come in over 7,000 unique flavors
 with and without nicotine, and while nicotine is a known neurotoxicant,
 the effects of vape flavoring alone, in the absence of nicotine, on brain
 function are not well understood. Here we performed a screen of vape
 aerosol extracts (VAEs) to determine the potential for prenatal
 neurotoxicity using the zebrafish embryo photomotor response (PMR) – a
 translational biosensor of neurobehavioral effects. We screened three
 commonly used aerosolized vape liquids (flavored and flavorless) either
 with or without nicotine. No neurobehavioral effects were detected in
 flavorless, nicotine-free VAEs, while the addition of nicotine to this VAE
 dulled sensory perception. Flavored nicotine-free VAEs also dulled sensory
 perception and caused hyperactivity in zebrafish embryos. The combination
 of flavor and nicotine produced largely additive effects. Flavored VAEs
 without nicotine had similar neuroactive potency as nicotine. Similar
 effects were also seen in embryos exposed to the pure flavoring compound,
 cinnamaldehyde. Together, using zebrafish PMR as a high throughput
 translational behavioral model for prenatal exposure, our results
 demonstrate that e-cigarette flavorants that we screened elicit
 neurobehavioral effects worthy of further investigation for long-term
 neurotoxic potential, and also have the potential to modulate nicotine
 impact on the developing brain.
description: The data were collected as described in the methods section of
 main article.
description: We include all data and provide R code reproduce the analysis.
 The included data are formated as .RData files and must be accessed
 through R. The script files (.R files) included contain all necessary code
 to access the data and repeat the analyses. Please make sure the .RData
 files are downloaded to the appropriate folder for assigning your
 workspace in R. pmr.fitB.me2.Rdata includes
 data for the blueraspberry vape trials
 pmr.fitC.me2.Rdata includes data for the cinnamon vape
 trials pmr.fitF.me2.Rdata includes data for the
 flavorless vape trials pmr.fit2.me2.Rdata includes
 data for the cinnamaldehyde trials VAEscript.R
 includes R code for replicating the analysis of the vape trails
 Cinnamaldehydescript.R includes R code for replicating the
 analysis of the cinnamaldehyde trials 
 Required R libraries are specified in the .R script files. Data
 are included as data.frame objects in R, with columns representing
 variables. Raw housekeeping variables remain in the
 data.frames.  Variables in the data.frames for vape
 trials (VAEscript.R) inlcude: v -&gt;
 time in seconds treatment -&gt; raw treatment
 labels (housekeeping) col -&gt; column ID on the
 96-well plate plate -&gt; 96-well plate
 ID x -&gt; raw % concentration
 labels (housekeeping) totaldist -&gt; raw
 activity conc -&gt; % vape aersol extract in
 exposure media ID -&gt; subject ID
 tx -&gt; raw treatment interaction ID
 labelling (housekeeping) nictoine -&gt; presence
 or absence ID for nicotine in vapes int -&gt;
 3-way treatment interation ID used for nlme analysis
 flavour -&gt; vape flavour 
 Variables in the data.frames for cinnamaldehyde trials
 (Cinnamaldehydescript.R) include: Time
 -&gt; raw time in seconds (housekeeping) well
 -&gt; well ID on the 96-well plate totaldist
 -&gt; raw activity plate -&gt; 96-well plate
 ID treatment -&gt; % concentration
 labels col -&gt; column ID on the 96-well
 plate x -&gt; raw treatment ID
 ID -&gt; subject ID v -&gt; time in
 seconds