The Zebrafish for Cardiovascular Studies In Vivo The aims of this platform are to provide EVGN scientists with a direct access to the zebrafish model system for in vivo studies of cardiovascular development and disease. The zebrafish is an ideal model for screening the angiogenic and cardiovascular regenerative properties of novel genes. This is achieved through the direct visualization of the vascular tree in transgenic lines expressing green fluorescent proteins (GFP or RFP) in developing and mature vessels and the use of morpholino knock down in the transgenic lines.

We use the tg(fli1-GFP) line (Lawson and Weinstein, 2002 see also: The Interactive Atlas of Zebrafish Vascular Anatomy), which has robust GFP expression in the cardiac and vascular endothelium and in leukocytes, starting from the earliest stages of vessel development through adulthood (see movie 1 and movie 2).
We perform two levels of assays, level one suited for high throughput and basic screening of knock down phenotypes, and level two for a thorough analysis of gene function in angiogenesis. The assays can be combined and customized.
 
Here are some examples:
 
Level 1- (loss of function/angiogenesis):
Screening the effects of gene knock down with specific morpholinos using the tg(fli1-GFP) line at 1,2 and 3 dpf (days post fertilization).  The screening will be carried out by direct visualization of the intersomitic vessels, (fig.1), cranial vasculature and cardiac endothelium.

Level 2- (gain of function/cardio-vascular integrity/regeneration):
To complement the loss of function approach, gain of function can be obtained through over expression. Constructs for over expression can also be used for rescue experiments. Additional levels of analysis of the phenotypes can be custom designed: i.e. focusing on cardiac endothelial integrity, vascular regeneration in the caudal fin assay, capillary integrity (leakage of fluorescent beads), etc

This platform is manage at IFOM by Marina Mione.

Download our leaflet here!