Single-cell transcriptome sequencing analysis of endothelial cells in control and high glucose treated zebrafish embryos

Artificially sweetened beverages containing noncaloric monosaccharides were suggested as healthier alternatives to sugar-sweetened beverages. Nevertheless, the potential detrimental effects of these noncaloric monosaccharides on blood vessel function remain inadequately understood. Presently, we have established a zebrafish model that exhibits significant excessive angiogenesis induced by high glucose. Utilizing this model, we observed that glucose and noncaloric monosaccharides could induce excessive formation of blood vessels, especially intersegmental vessels (ISVs). The excessively branched vessels were observed to be formed by ectopic activation of quiescent endothelial cells (ECs) into tip cells. Single-cell transcriptomic sequencing analysis of the endothelial cells in the embryos exposed to high glucose revealed an augmented ratio of capillary ECs, proliferating ECs, and a series of upregulated proangiogenic genes. Overall design: Firstly, we isolated the EGFP-positive cells from control and high glucose-treated Tg(fli1ep:EGFP-CAAX) transgenic embryos. Following the proteolytic dissociation of embryos, the EGFP-positive cells were isolated by fluorescence-activated cell sorting (FACS). Around 300~500 zebrafish embryos were used for the ECs collection for each stage. The isolated ECs were analyzed using a large-scale scRNA-seq (10X Genomics) platform.