Intracerebral haemorrhage (ICH) in zebrafish larvae results in a quantifiable brain injury. (A) Representative images of the brain injury phenotype in ICH+ larvae (right panels), in comparison to ICH- siblings (left panels), at 72 hpf. Bright-field images (bottom panels) demonstrate the presence of brain bleeds (arrows) in ICH+ larvae. Fluorescent microscopy was performed to visualise cell death in the ubiq:secAnnexinV-mVenus reporter line (top panels). Clusters of dying cells were observed in peri-haematomal regions. Images were cropped to brain only regions and analysed for total green fluorescence intensity in round particles bigger than 30 pixels in diameter (white line). (B) Quantification of fluorescent signal in the brains of untreated, ICH- and ICH+ larvae obtained through the ATV model (n=12 per group; 3 independent replicates) at 72 hpf. Significant differences were observed when comparing ICH+ with untreated (**p=0.004) and with ICH- (*p=0.03) siblings. (C) Quantification of fluorescent signal as a read out for annexinV binding in the brains of ICH- and ICH+ larvae obtained through the bubblehead (bbh) model (n=12 per group; 2 independent replicates) at 72 hpf. A significant difference in mVenus fluorescence was observed between ICH+ and ICH- age-matched siblings (**p=0.002). Original magnification, x20.
|
