title: Characterization of P2X7R and its function in the macrophages of ayu, Plecoglossus altivelis.
abstract: P2X purinoceptor 7 (P2X7R), an ATP-gated ion channel, plays an important role during the innate immune response in mammals. However, relatively little is known about the role of P2X7R in the fish immune system. Here, we cloned a cDNA sequence encoding ayu (Plecoglossus altivelis) P2X7R (aP2X7R). The predicted protein was composed of 574 amino acid residues with a P2X family signature, two transmembrane domains, and a long C-terminal. aP2X7R transcripts were mainly distributed in ayu immune tissues and significantly increased in all tested tissues and in macrophages after Listonella anguillarum infection. The aP2X7R protein was upregulated significantly in macrophages upon bacterial challenge. An antibody against the ectodomain of aP2X7R (aEPAb) and an antagonist (oATP) were used to block aP2X7R. aP2X7R siRNA was also used to knockdown the receptor expression in ayu macrophages. Cell death induced by ATP was significantly inhibited in ayu macrophages after aEPAb, oATP, or siRNA treatment. Moreover, aP2X7R ablation also resulted in suppression of phagocytic activity and ATP-induced bacterial killing in ayu macrophages. Our results indicated that aP2X7R was upregulated after infection and mediated cell death, phagocytosis, and bacterial killing of ayu macrophages.
authors: School of Marine Sciences, Ningbo University, Ningbo, China.
authors: Yu-Qing
authors: YQ
authors: He
authors: Jiong
authors: J
authors: Chen
authors: Xin-Jiang
authors: XJ
authors: Lu
authors: Yu-Hong
authors: YH
authors: Shi
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D057134:Antibodies, Neutralizing / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005393:Fish Diseases; D029941:Fish Proteins / Q000235:genetics* / Q000276:immunology; D005786:Gene Expression Regulation / Q000187:drug effects; D016905:Gram-Negative Bacterial Infections / Q000276:immunology* / Q000382:microbiology / Q000662:veterinary*; D053526:Listonella / Q000254:growth & development; D008264:Macrophages / Q000187:drug effects / Q000276:immunology* / Q000382:microbiology; D008969:Molecular Sequence Data; D036441:Osmeriformes / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D010587:Phagocytosis / Q000187:drug effects; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D034741:RNA, Small Interfering / Q000235:genetics; D058486:Receptors, Purinergic P2X7 / Q000235:genetics* / Q000276:immunology; D016415:Sequence Alignment
chemical_list: D057134:Antibodies, Neutralizing; D018076:DNA, Complementary; D029941:Fish Proteins; D034741:RNA, Small Interfering; D058486:Receptors, Purinergic P2X7; C017199:2',3'-dialdehyde ATP; D000255:Adenosine Triphosphate