Bu, Fan

Jiang, Liwen

Ma, Dongqing

Quinn, John P

Wang, Minyan

University of Liverpool Repository

Electrophysiological technique was performed to detect the effect of anti-P2X7R antibody on CSD.  Real time PCR was then performed to analyze the gene level of IL-1β and TNF-α post CSD with or without antibody treatment The anti-P2X7R antibody suppressed CSD and CSD-induced TNF-α gene expression Dongqing Maabc , Fan Buabc , Liwen Jiangabc , John P Quinnc , Minyan Wangab a Centre for Neuroscience, b Department of Biological Sciences, Xi’an Jiaotong-Liverpool University, Suzhou, 215123, China;     c Department of Molecular and Clinical Pharmacology, Institute of Translational Medicine, University of Liverpool, Liverpool, UK  •  The purinergic P2X7 receptors, are ionotropic membrane channels, which can be activated by extracellular ATP. •  ATP is known to be involved cortical spreading depression (CSD), a substrate of migraine [1, 2] . •  P2X7R activation promotes IL-1β[3] and TNF-α[4] release and CSD triggers these neuroinflammatory factors release[5] . •  Inhibition of the P2X7-Panx1 complex suppresses spreading depolarization in rats and the induction of IL-1β post CSD [6]  •  Whether the anti-P2X7R antibody could suppress CSD and the subsequent induction of neuroinflammatory factors is unknown. •  To examine the effects of anti-P2X7R antibody on cortex susceptibility to CSD. •  To address whether P2X7R contributes to the induction of IL-1β and TNF-α induced by CSDResults: References: 1. BURNSTOCK, G. & RALEVIC PHARMACOL REV, 66, 102-92., 2014. 2. SCHOCK, S.C. ET AL., BRAIN RES, 1168, 129-38. 3 VERHOEF, P. A., ET AL., J IMMUNOL, 170, 5728-38. 2003. 4. SUZUKI, T, ET AL., J NEUROSCI, 24, 1-7. 2004. 5. JANDER, S. ET AL., J CEREB BLOOD FLOW METAB, 21, 218-25. 2001. 6. CHEN, S.P. ET AL., BRAIN, 2017. First author information:   dongqing.ma13@xjtlu.edu.cn, the final year PhD student in XJTLU and UoL. Acknowledgment: Supervisors: Dr. Minyan Wang and Prof. John P Quinn; Yan Wang; Funding from XJTLU and Wenli Wang charitable foundation. (ii) CSD recording (i)  CSD induction by 2 M KCl topically perfused on cortex with dura intact (iii) Antibody intracerebroventricular  (i.c.v.) injection Anterior Posterior BregmaFigure 1. Schematic representation of surgical coordinates on parietal bone of isoflurane anaesthetic rats. Figure 2. Representative trace in DC potential of CSD wave. Latency: the time required to elicit the 1st CSD wave from the start of high K+ application. Figure 3. Timeline of the in vivo experiment with sequence of pharmacological and electrophysiological performance followed by tissue dissection for the subsequent molecular analysis. •  Blockade of  P2X7R by antibody reduced cortex susceptibility to CSD and rapidly reduced CSD induced gene expression of TNF-α in rats, indicating P2X7R as a potential target for migraine prophylaxis and treatment. •  The anti-P2X7R antibody did not reduce the induction of gene expression of IL-1β immediately after CSD. This may be due to: 1) the time points, as a significant down-regulation of IL-1β mRNA level was reported by inhibition of the P2X7/PANX1 pore complex 4 hours post CSD[6]. 2) suppression of spreading depolarization-induced IL-1β mRNA upregualtion was due to inhibition of the P2X7/PANX1 pore complex, not the P2X7R cation channel [6]. 1. Anti-P2X7R antibody suppressed CSD in ratsFigure 5. gene expression of TNF-α and IL-1β, (n=5 in glycerol sham group and n=8 in another two group). *p < 0.05, **p < 0.01, ****p<0.0001. Two-tailed unpaired t-test. Figure 4. The anti-P2X7R antibody reduced cortex susceptibility to CSD in rats. (n=8 in each group) *p < 0.05, **p < 0.01. Two-tailed, unpaired t-test. Methods: Objectives: Background: Conclusion and Further Work: 2. Anti-P2X7R antibody reduced the induction of  mRNA of  TNF-α, but not IL-1β by CSDK+ application ▪ ▪ ▪▪ ▪CSD numberLatency 5 min 10 mV (-)10 min            3 days                            1 hour                 30 min  10 min (day 0) Anti-P2RX7 Ab i.c.v. (0.5 µl / min) Recovery (day 4) Surgery for CSD recording CSD recording 2 M KCl for CSD induction 

P2X7 receptor regulates CSD and CSD-induced TNF-α induction

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P2X7 receptor regulates CSD and CSD-induced TNF-alpha induction

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