Department of Pathobiological Sciences, University of Wisconsin-Madison, Madison, WI, USA.
Department of Biological Sciences, Clemson University, Clemson, SC, USA.
Sci Rep. 2020 Sep 24;10(1):15716. doi: 10.1038/s41598-020-72750-9.
Tissue damage induces rapid recruitment of leukocytes and changes in the transcriptional landscape that influence wound healing. However, the cell-type specific transcriptional changes that influence leukocyte function and tissue repair have not been well characterized. Here, we employed translating ribosome affinity purification (TRAP) and RNA sequencing, TRAP-seq, in larval zebrafish to identify genes differentially expressed in neutrophils, macrophages, and epithelial cells in response to wounding. We identified the complement pathway and c3a.1, homologous to the C3 component of human complement, as significantly increased in neutrophils in response to wounds. c3a.1 zebrafish larvae have impaired neutrophil directed migration to tail wounds with an initial lag in recruitment early after wounding. Moreover, c3a.1 zebrafish larvae have impaired recruitment to localized bacterial infections and reduced survival that is, at least in part, neutrophil mediated. Together, our findings support the power of TRAP-seq to identify cell type specific changes in gene expression that influence neutrophil behavior in response to tissue damage.
组织损伤诱导白细胞的快速募集和转录谱的变化，从而影响伤口愈合。然而，影响白细胞功能和组织修复的细胞类型特异性转录变化尚未得到很好的描述。在这里，我们在幼虫斑马鱼中采用翻译核糖体亲和纯化（TRAP）和 RNA 测序（TRAP-seq）来鉴定在中性粒细胞、巨噬细胞和上皮细胞中响应伤口而差异表达的基因。我们发现补体途径和 c3a.1，与人类补体的 C3 成分同源，在伤口反应中中性粒细胞中显著增加。c3a.1 斑马鱼幼虫对尾部伤口的中性粒细胞定向迁移受损，在受伤后早期招募时出现初始滞后。此外，c3a.1 斑马鱼幼虫对局部细菌感染的募集减少，存活率降低，至少部分是由中性粒细胞介导的。总之，我们的研究结果支持 TRAP-seq 能够识别影响组织损伤后中性粒细胞行为的细胞类型特异性基因表达变化。
