Developmental and comparative study of hematopoiesis in the lamprey: insights into the evolutionary origin of hematopoiesis in vertebrates

七鳃鳗造血作用的发育和比较研究：深入了解脊椎动物造血作用的进化起源

• 批准号: 19K06798
• 负责人: PASCUAL JUAN
• 金额: $ 2.83万
• 依托单位: Institute of Physical and Chemical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2019
• 资助国家: 日本
• 起止时间: 2019-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-19K06798/
• 关键词: hematopoiesis; vertebrates; evolution; evo-devo; Evo-Devo; lamprey

Hematopoiesis is the process by which the blood cells of an individual, from the embryo to adulthood, are formed. However, despite being a crucial physiological phenomenon ofanimals, most of our knowledge derives from studies using a handful of classic animal models, mostly representatives of jawed vertebrates. Withthis in mind, I set to study the lamprey, long-distant relative of jawed vertebrates that is understudied in terms of the blood system. In FY2020 I completed the analysis of RNA-seq data from lamprey embryos and identified all candidate hematopoietic markers in the lamprey. I generated a complete panel of expression patterns for all putative hematopoietic markers during lamprey embryogenesis. I was then able to perform CRISPR knock-outs of lamprey Scl gene, an important hematopoietic factor in other vertebrates, and obtained enough samples to carry out RNA-seq analysis as well as in situ hybridization experiments. Although there was no apparent mutant phenotype, blood alterations are expected. These samples will be used in the near future to assess the function of Scl in blood development in cyclostomes, what will enable comnparative analysis with gnathostome hematopoiesis.

造血是个体的血细胞从胚胎到成年形成的过程。然而，尽管这是动物的一个重要生理现象，但我们的大部分知识都来自于使用少数经典动物模型的研究，其中大多数是有颌脊椎动物的代表。考虑到这一点，我开始研究七鳃鳗，它是有颌脊椎动物的远亲，在血液系统方面研究得不够。在2020财政年度，我完成了对七鳃鳗胚胎RNA-seq数据的分析，并确定了七鳃鳗中所有候选的造血标志物。我产生了一个完整的面板的表达模式，所有假定的造血标记在七鳃鳗胚胎发生。然后，我能够对七鳃鳗Scl基因（其他脊椎动物中重要的造血因子）进行CRISPR敲除，并获得足够的样本进行RNA-seq分析以及原位杂交实验。虽然没有明显的突变表型，但预期会发生血液变化。这些样本将在不久的将来用于评估Scl在圆口动物血液发育中的功能，这将使得能够与颚口动物造血进行比较分析。