Figure 4
Analysis of lipid profile with BMAA170 treated embryo. Intact 96 hpf embryos were homogenized to separate lipid and are distributed in two parts. One part is dried and used for HPTLC and MALDI-ToF analysis of lipids and other part is suspended in 100 mM phosphate buffer and distributed in 96 well plates for cholesterol assay. (A) Reaction diagram of the enzyme activity assay of cholesterol measured photometrically at 590 ± 10 nm. (B) Quantification of cholesterol using Amplex red agent based enzymatic assay lit. Significant reduction of cholesterol (p < 0.001; n = 9) in BMAA170 treated embryo is observed. (C) Image of a HPTLC plate of a reference PL mixture (1.69 mg each) and the embryo lipid extracts. (D) Comparison of negative ion MALDI-ToF lipid profiles in BMAA170-exposed zebrafish embryos, relative to controls. Phosphatidylinositol (PI) lipids are decreased and phosphatidylethanolamine (PE) lipids are increased in BMAA170 treated embryos compared to controls.
