Transcriptome analysis of wt1b:eGFP positive macrophages in the regenerating zebrafish heart

Little is known about zebrafish macrophage subtypes and their contribution to organ regeneration. Using the transgenic line wt1b:eGFP we identified a subtype of macrophages in the regenerating heart that is transcriptionally different from the rest of macrophages. Hearts from Tg(wt1b:eGFP; mpeg1:mCherry) zebrafish were cryoinjured and at 4 dpi mCherry and GFP/mCherry cells were FAC sorted. RNA was extracted from 4 pools of GFP/mCherry and 4 pools of mCherry single positive cells and RNAseq was performed.

目前学界对斑马鱼巨噬细胞亚型及其在器官再生中的调控作用尚缺乏系统认知。本研究利用转基因品系wt1b:eGFP，在再生过程中的心脏组织内鉴定出一类转录谱与其余巨噬细胞存在显著差异的巨噬细胞亚型。研究人员对Tg(wt1b:eGFP; mpeg1:mCherry)转基因斑马鱼的心脏实施冷冻损伤，并于损伤后第4天（days post injury, dpi）通过荧光激活细胞分选（Fluorescence-activated cell sorting, FACS）分离得到mCherry单阳性细胞与GFP/mCherry双阳性细胞。随后从4组GFP/mCherry双阳性细胞混合样本以及4组mCherry单阳性细胞混合样本中提取总RNA，开展RNA测序（RNA-seq）实验。