ZeEfficacy – Efficacy Services

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Atherosclerosis Model

Atherosclerosis is a chronic inflammatory disease driven in large part by hypercholesterolemia. It is caused by the buildup of fat, cholesterol and other substances within the arteries and on their walls. This buildup, called plaque, can cause the arteries to narrow, obstructing blood flow and leading to other complications such as myocardial infarction or stroke.

Human fat metabolism is highly conserved in zebrafish. Indeed, zebrafish express all main nuclear receptors, lipid transporters, apolipoproteins and enzymes involved in lipoprotein metabolism. Even the activity of the cholesteryl ester transfer protein (CETP), which is naturally deficient in rodents, is detected in zebrafish plasma. 

Feeding zebrafish a High Cholesterol Diet (HCD) results in significant hypercholesterolemia and initiation of vascular processes, which configures zebrafish as an attractive model to study mechanisms relevant to early development of human atherosclerosis.

We offer a fast, cost-effective and biologically-relevant model for the study of cholesterol metabolism and atherosclerosis condition with a diet-induced approach.

Applications

  • Study the effect of nutritional interventions on cholesterol accumulation and development of atherosclerosis.
  • Perform gene candidate screening for cholesterol metabolism and atherosclerosis.
  • Screening of potential cholesterol-regulatory molecules.

Advantages

Lipid and lipoprotein metabolism in zebrafish and in humans are remarkably similar.

Transparent zebrafish larvae facilitate in vivo cholesterol quantification via non-invasive confocal imaging.

Vascular processes can be monitored in live animals.

High throughput screening of molecules.

Method description

Early-life embryos (5th dpf) are fed a Non-Cholesterol Diet (NCD) versus a HCD for 15 days in the presence and absence of the compound of interest. Individuals are posteriorly incubated with Bodipy fluorescent dye for the in vivo confocal visualization of cholesterol.

Readouts

  • Whole body cholesterol levels: treated larvae are processed via enzymatic colorimetric kits.
  • Vascular cholesterol accumulation: treated larvae are processed via Bodipy labelling.
Figure 1. Vascular cholesterol accumulation. Colocalization analysis of flk1:gfp trangenic zebrafish expressing green fluorescent protein (GFP) in the vasculature and cholesterol labelled with Bodipy dye (red fluorescence). From left to right: normal control diet (NCD) and high-cholesterol diet (HCD) fed zebrafish, and finally zebrafish fed with HCD plus an active compound, that does not efficiently reduce cholesterol accumulation.
Figure 2. Whole body cholesterol measurements of normal control diet (NCD) and high-cholesterol diet (HCD) fed zebrafish, and zebrafish fed with HCD plus an active compound, that does not efficiently reduce cholesterol accumulation.