Fig. 3

Co-exposure to alcohol and benzo[a]pyrene-induced membrane remodeling in the liver of HFD zebrafish larvae. Membrane order and lipid raft spatial distribution characteristics of membrane remodeling was assessed in liver cells of steatotic zebrafish larvae after co-exposure to ethanol and B[a]P for seven days from 5 to 12 dpf. Zebrafish larvae under two conditions—untreated (C) or treated with combination of 43 mM ethanol and 25 nM B[a]P (BE)—were stained with di-4-ANEPPDHQ—a membrane order-sensitive fluorescent probe—and analyzed by confocal fluorescence microscopy. Membrane order in membranes of zebrafish liver was measured by computing the generalized polarization (GP) factor. (A) Changes in GP values were expressed as the difference between individual larva GP value and the mean of GP found in control larvae (ΔGP). (B) On the left, some representative liver images of each treatment have been selected according to the respective mean of delta GP (magnification ×400). Pixels with higher GP values (which could be considered as lipid rafts) have been highlighted in yellow to pinpoint lipid raft spatial distribution. The liver area outlined in the white square on the left images are magnified on the right side to show lipid raft spatial distribution in the plasma membrane. Values are the mean ± standard error of the mean (SEM) of at least 25 larvae. * Significantly different from HFD control larvae.