Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal University, 393 West Binshui Road, Xiqing District, Tianjin 300387, China.
Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal University, 393 West Binshui Road, Xiqing District, Tianjin 300387, China.
Fish Shellfish Immunol. 2018 Apr;75:208-215. doi: 10.1016/j.fsi.2018.02.020. Epub 2018 Feb 9.
G-protein-coupled P2Y receptors activated by extracellular nucleotides play important roles under different physiological and pathophysiological conditions in mammals. To investigate the immunological relevance of P2Y receptors in fish, we identified and characterized the P2Y and P2Y receptors in Japanese flounder Paralichthys olivaceus. The P. olivaceus P2Y and P2Y receptors harbor seven transmembrane domains but share only 24% sequence identity. Real-time PCR analysis revealed the constitutive but unequal mRNA expression pattern of P2YR and P2YR in normal Japanese flounder tissues with the dominant expression of P2YR in head kidney and blood and P2YR in hepatopancreas. In addition, the expression of P2Y and P2Y receptors was markedly modulated by PAMPs stimulation and Edwardsiella tarda infection. Furthermore, blockage of P2YR potently increased ADP-activated pro-inflammatory cytokine IL-1beta gene expression in the head kidney macrophages (HKMs). Moreover, inhibition of P2Y and P2Y receptor activity with their respective potent antagonists significantly altered some of the LPS-induced pro-inflammatory cytokine gene expression in the HKMs. However, blockade of P2YR did not affect the poly(I:C)-induced pro-inflammatory cytokine gene expression examined in the HKMs. Collectively, we have for the first time reported the role of purinergic P2Y and P2Y receptors in fish innate immunity. Our findings have also addressed the importance of extracellular ATP and its metabolites in fish innate immune responses.
G 蛋白偶联的 P2Y 受体被细胞外核苷酸激活，在哺乳动物的不同生理和病理生理条件下发挥重要作用。为了研究嘌呤能 P2Y 受体在鱼类中的免疫学相关性，我们鉴定并表征了日本牙鲆 Paralichthys olivaceus 的 P2Y 和 P2Y 受体。日本牙鲆的 P2Y 和 P2Y 受体具有七个跨膜结构域，但序列同一性仅为 24%。实时 PCR 分析显示，正常日本牙鲆组织中 P2YR 和 P2YR 的 mRNA 表达模式呈组成型但不均等，头肾和血液中 P2YR 的表达占主导地位，而 P2YR 在肝胰脏中表达。此外，PAMP 刺激和迟缓爱德华氏菌感染显著调节 P2Y 和 P2Y 受体的表达。此外，P2YR 的阻断强烈增加了头肾巨噬细胞（HKMs）中 ADP 激活的促炎细胞因子 IL-1beta 基因的表达。此外，用各自的有效拮抗剂阻断 P2Y 和 P2Y 受体活性显著改变了 HKMs 中 LPS 诱导的一些促炎细胞因子基因的表达。然而，阻断 P2YR 并没有影响 HKMs 中检测到的 poly(I:C)诱导的促炎细胞因子基因的表达。总之，我们首次报道了嘌呤能 P2Y 和 P2Y 受体在鱼类先天免疫中的作用。我们的研究结果还强调了细胞外 ATP 及其代谢物在鱼类先天免疫反应中的重要性。
