Abstract:
Larval zebrafish (Danio rerio), 48-96 hours post-fertilization (hpf), were assessed for visual and cardiac function using molecular, immunohistological, physiological, and behavioral endpoints. An optokinetic response was determined using a rotating drum apparatus. Behavioral tests were video recorded and analyzed for mean eye saccades for 96 hpf larvae, with videos recorded for 48 hpf larvae to assess heart rate, stroke volume, and cardiac output. Larvae were imaged for total length and eye diameter measurements and subsequently preserved for immunohistological and gene expression analysis. This dataset supports the publication: Magnuson, J. T., Bautista, N. M., Lucero, J., Lund, A. K., Xu, E. G., Schlenk, D., Burggren, W.W., & Roberts, A. P. (2020). Exposure to Crude Oil Induces Retinal Apoptosis and Impairs Visual Function in Fish. Environmental Science & Technology, 54(5), 2843–2850. doi:10.1021/acs.est.9b07658.
Suggested Citation:
Magnuson, Jason, Edward Mager and Aaron Roberts. 2020. Dataset for: Exposure to Crude Oil Induces Retinal Apoptosis and Impairs Visual Function in Fish. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N78W3BX7
Purpose:
To determine the impact of oil exposure on retinal development in teleost fish (zebrafish; Danio rerio) using molecular, immunohistological, physiological, and behavioral endpoints.
Data Parameters and Units:
The dataset contains both raw data (tiff files and videos) and processed data used to plot graphs in the paper. Days post fertilization (dpf); high energy water accommodated fraction (HEWAF); 0, 1, 3, 5, and 7 percent relates to the percent of diluted HEWAF. Images were analyzed using ImageJ software (Version 1.47); Standard length and eye diameters were measured in micrometers in 96 hpf; Larvae assessed for cardiovascular analysis were measured for standard length in millimeters, heart rate (fH), stroke volume (Vs), and cardiac output (Q); Stroke volume was measured in nano-liters times beats per minute (bpm), whereas cardiac output was measured in nano-liters times minutes.
Methods:
Larval zebrafish (Danio rerio), 96 hours post fertilization (hpf), were used. Zebrafish were embryonically exposed to 0, 1, 3, 5, or 7 percent high energy water accommodated fractions (HEWAFs), with each treatment run with 5 replicates and each replicate containing 30 embryos. Fish were recorded under a Nikon SMZ800 stereoscope, with videos recording eye saccades at 100 frames/second. Eye saccades were counted in both clockwise and counterclockwise directions, with a mean total saccade used as an endpoint. Files were named as percent HEWAF exposure followed by the individual fish assessed and the subsequent replicate used. For example, (1-2) represents a zebrafish exposed to a 1 percent HEWAF from replicate dish 2.
Immunohistochemical analysis:
Image names are labeled according to percent HEWAF exposure (0, 1, 3, 5, and 7 percent HEWAF), the image number, the magnification the image was taken at, and the image overlay. Four types of overlays were taken: RFP, GFP, DAPI, and RGB. RFP-red fluorescent protein fluoresced for apoptotic activity, GFP- green fluorescent protein fluoresced for Müller glial cell activity, DAPI- fluoresced for Hoechst nuclear stains, and RGB is the overlay of RFP, GFP, and DAPI. Data are represented as corresponding fluorescent units, with all images analyzed using ImageJ software (version 1.47).
Quantitative polymerase chain reaction (qPCR) analysis:
All qPCRs were run on a Rotor-Gene 6000, with outputs recorded as Ct¬ values for zebrafish exposed to 0, 1, 3, 5, or 7 percent HEWAF.
Morphometric and cardiovascular measurements:
Images are denoted as hours post fertilization (hpf), concentration and accompanying replicate number, and fish number analyzed. Standard length and eye diameters were measured in micrometers in 96 hpf. Larvae assessed for cardiovascular analysis were measured for standard length in millimeters, heart rate (fH), stroke volume (Vs), and cardiac output (Q). Stroke volume was measured in nano liters times beats per minute (bpm), whereas cardiac output was measured in nano liters multiplied by minutes.
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