Transcriptomic Evidence Reveals the Molecular Basis for Functional Differentiation of Hemocytes in a Marine Invertebrate,Crassostrea gigas
Transcriptomic Evidence Reveals the Molecular Basis for Functional Differentiation of Hemocytes in a Marine Invertebrate,Crassostrea gigas
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转录组学证据揭示了海洋无脊椎动物巨牡蛎血细胞功能分化的分子基础
DOI：
10.3389/fimmu.2020.00911
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发表时间：
2020-05-27
影响因子：
7.3
通讯作者：
Yu, Ziniu
中科院分区：
文献类型：
--
作者：
Mao, Fan;Wong, Nai-Kei;Yu, Ziniu
血细胞在双壳类软体动物的宿主免疫防御中发挥着明确的核心作用，尽管其功能分化的确切机制仅部分了解。为此，通过流式细胞术从太平洋牡蛎的血细胞中分选出粒细胞和透明细胞，因此定量转录组学分析揭示了一系列显著的差异表达基因（DEG），这些基因在粒细胞中全面上调，致力于牡蛎血细胞之间的功能分化。我们的DEG网络说明了积极参与的信号传导途径，Cdc 42/Cdc 42 l是途径网络的核心调节因子，这通过Cdc 42抑制剂存在下血细胞吞噬能力的显著降低来验证。此外，在DEG中鉴定了许多转录因子，包括ELK、HELT和Fos，它们主要在粒细胞中表达。AP-1转录因子Fos在通过AP-1结合位点与靶基因结合的测定中被证实促进血细胞的功能分化，与下游吞噬作用和ROS产生一致。重要的是，Cdc 42/Cdc 42 l也受到Fos表达的调控，提供了可能的调控机制-引导血细胞功能分化。这项研究的发现填补了海洋无脊椎动物血细胞功能分化机制的知识空白。gigas，为研究吞噬细胞在更高级和更新的门中的免疫防御进化和功能分化奠定了基础。
Hemocytes play unequivocally central roles in host immune defense of bivalve mollusks, though the exact mechanisms underlying their functional differentiation are only partially understood. To this end, granulocytes and hyalinocytes were sorted via flow cytometry from hemocytes of the Pacific oysterCrassostrea gigas, and consequently quantitative transcriptomic analysis revealed a striking array of differentially expressed genes (DEGs), which were globally upregulated in granulocytes, dedicating to functional differentiation among oyster hemocytes. Our network of DEGs illustrated actively engaged signaling pathways, with Cdc42/Cdc42l being a core regulator of pathway network, which was validated by a dramatically reduced capacity for hemocyte phagocytosis in the presence of Cdc42 inhibitors. Additionally, a number of transcription factors were identified among DEGs, including ELK, HELT, and Fos, which were predominantly expressed in granulocytes. The AP-1 transcription factor Fos was confirmed to facilitate functional differentiation of hemocytes in an assay on binding to target genes by the AP-1 binding site, consistent with downstream phagocytosis and ROS production. Importantly, Cdc42/Cdc42l were also regulated by the expression of Fos, providing a possible regulatory mechanism-guided hemocyte functional differentiation. Findings in this study have bridged a knowledge gap on the mechanistic underpinnings of functional differentiation of hemocytes in a marine invertebrateC. gigas, which promise to facilitate research on the evolution of immune defense and functional differentiation of phagocyte in higher-order and more recent phyla.