Department of Biological Sciences, University of North Texas, Denton, TX, United States of America.
BioDiscovery Institute, University of North Texas, Denton, TX, United States of America.
PLoS One. 2022 Mar 23;17(3):e0264776. doi: 10.1371/journal.pone.0264776. eCollection 2022.
The zebrafish is an excellent model system to study thrombocyte function and development. Due to the difficulties in separating young and mature thrombocytes, comparative transcriptomics between these two cell types has not been performed. It is important to study these differences in order to understand the mechanism of thrombocyte maturation. Here, we performed single-cell RNA sequencing of the young and mature zebrafish thrombocytes and compared the two datasets for young and mature thrombocyte transcripts. We found a total of 9143 genes expressed cumulatively in both young and mature thrombocytes, and among these, 72% of zebrafish thrombocyte-expressed genes have human orthologs according to the Ensembl human genome annotation. We also found 397 uniquely expressed genes in young and 2153 uniquely expressed genes in mature thrombocytes. Of these 397 and 2153 genes, 272 and 1620 corresponded to human orthologous genes, respectively. Of all genes expressed in both young and mature thrombocytes, 4224 have been reported to be expressed in human megakaryocytes, and 1603 were found in platelets. Among these orthologs, 156 transcription factor transcripts in thrombocytes were found in megakaryocytes and 60 transcription factor transcripts were found in platelets including a few already known factors such as Nfe2 and Nfe212a (related to Nfe2) that are present in both megakaryocytes, and platelets. These results indicate that thrombocytes have more megakaryocyte features and since platelets are megakaryocyte fragments, platelets also appear to be thrombocyte equivalents. In conclusion, our study delineates the differential gene expression patterns of young and mature thrombocytes, highlighting the processes regulating thrombocyte maturation. Future knockdown studies of these young and mature thrombocyte-specific genes are feasible and will provide the basis for understanding megakaryocyte maturation.
斑马鱼是研究血小板功能和发育的优秀模型系统。由于难以分离年轻和成熟的血小板，因此尚未对这两种细胞类型进行比较转录组学研究。研究这些差异对于了解血小板成熟的机制非常重要。在这里，我们对年轻和成熟的斑马鱼血小板进行了单细胞 RNA 测序，并比较了这两个数据集的年轻和成熟血小板转录本。我们总共发现了 9143 个在年轻和成熟血小板中均表达的基因，其中根据 Ensembl 人类基因组注释，72%的斑马鱼血小板表达基因具有人类同源基因。我们还发现了 397 个在年轻血小板中特异表达的基因和 2153 个在成熟血小板中特异表达的基因。在这 397 个和 2153 个基因中，分别有 272 个和 1620 个对应于人类同源基因。在所有在年轻和成熟血小板中表达的基因中，有 4224 个已被报道在人类巨核细胞中表达，有 1603 个在血小板中发现。在这些同源基因中，在巨核细胞和血小板中发现了 156 个血小板转录因子的转录本，包括一些已经知道的因子，如 Nfe2 和 Nfe212a（与 Nfe2 相关），它们存在于巨核细胞和血小板中。这些结果表明血小板具有更多的巨核细胞特征，由于血小板是巨核细胞的碎片，因此血小板似乎也与血小板等效。总之，我们的研究描绘了年轻和成熟血小板的差异基因表达模式，突出了调节血小板成熟的过程。对这些年轻和成熟血小板特异性基因的未来敲低研究是可行的，这将为理解巨核细胞成熟提供基础。
