Figure 5

Adipose Deposition in Zebrafish Developmentally Exposed to Nonylphenol and Polyethoxylates. Zebrafish were developmentally exposed to control chemicals, nonylphenol, or nonylphenol polyethoxylates; aged to 30 days post-fertilization; then measured and stained with Nile Red (0.5 μg/mL) for 30 min. Total body fluorescence (A) was imaged at 16× magnification using a yellow fluorescent protein filter (representative images in Figure 6) and fluorescence was quantified for each fish and then biological replicates were averaged. n = 24 (DMSO), 16, 21, 25, 15, 20, 24, 14, 21, 25, 15, 25, 27, 22, 28, 25, 22, 27, 26, 25, 23, 26, and 25 across four spawning events (biological replicates) for exposure groups listed below, respectively. * indicates significant increase in total body fluorescence quantification over vehicle control fish, p < 0.05, as per Kruskal–Wallis test with Dunn’s multiple comparisons. # represents p < 0.10, as per statistics described above. Box and whisker plots depict the following metrics: whiskers represent 10–90th percentiles, box bounds represent the 25th to 75th percentiles, and the middle line represents the median. Developmental trajectory of adipose depots across the zebrafish (B). Relative proportions of fish exhibiting fluorescing adipocytes in each depot in the DMSO fish were set as “normal” and the heat map depicts increased proportions of fish in each group with visible adipocytes in each depot. Adipose depots labeled as per the developmental guides provided in Minchin and Rawls, 2017 (PMID: 28348140) and grouped based on anatomical classifications. A value of 9 represents a 9X increase in the proportion of fish in an exposure group PVAT = pancreatic visceral adipose tissue; AVAT = abdominal visceral adipose tissue; RVAT = renal visceral adipose tissue; aCVAT = anterior cardiac visceral adipose tissue; cPOS = central paraosseal non-visceral adipose tissue; OES = esophageal non-visceral adipose tissue; LSAT = lateral truncal adipose tissue; pOCU = posterior ocular adipose tissue; aOCU = anterior ocular adipose tissue; vOCU = ventral ocular adipose tissue; BHD = basihyal hyoid adipose tissue; CHD = ceratohyal hyoid adipose tissue; vOPC = ventral opercular adipose tissue; dOPC = dorsal opercular adipose tissue; and UHD = urihyal hyoid adipose tissue. DMSO = dimethylsulfoxide, vehicle control; TBT = tributyltin chloride; NPEO = nonylphenol polyethoxylated (with varying average ethoxylate chain lengths).