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. Author manuscript; available in PMC: 2013 Mar 11.
Published in final edited form as: Methods Cell Biol. 2011;101:111–141. doi: 10.1016/B978-0-12-387036-0.00005-0

Fig. 2.

Fig. 2

BODIPY lipid analogs enable studies of yolk metabolism during early zebrafish development. To assay the function of apoc2 during zebrafish development, embryos were injected with an apoc2 morpholino at the 1–4 cell stage followed by injection of a fluorescent fatty acid (BODIPY-C12) at 24 h postfertilization (hpf). (A) At 48 hpf, embryos injected with BODIPY-C12 retain the fluorescent analog primarily in their yolk. (B, C) apoc2 morphants exhibit an enlarged yolk phenotype (arrowhead) at 48 and 72 hpf, indicating that apoc2 is necessary for yolk utilization during larval development. (D, E) To determine the metabolic consequences of apoc2 deficiency, 1 dpf wild-type and apoc2 morphant larvae were injected with BODIPY-C12 and assayed using fluorescent thin layer chromatography (TLC) 2 days later. (D) TLC analysis shows that BODIPY-C12 is incorporated primarily into triacylglycerol (TAG), diacylglycerol (DG), and phosphatidylcholine (PC) in both wild-type and apoc2 morphant larvae. (E) apoc2 morphants exhibit defects in PC and lysophosphatidylcholine (LPC) metabolism. Total lipid fluorescence was quantified from TLC plates run with total lipids extracted from wild-type and apoc2 morphants. Triacylglycerol (TAG), diacylglycerol (DG), BODIPY C12:0 (C12), phospholipids (PL), PC, and LPC.* p < 0.05.