Department of Biological Sciences, Centre for Neuroscience, Xi'an Jiaotong-Liverpool University (XJTLU), Suzhou, 215123, China.
Department of Molecular and Clinical Pharmacology, Institute of Translational Medicine University of Liverpool, Liverpool, L69 7ZB, UK.
Int J Mol Sci. 2020 Feb 13;21(4):1269. doi: 10.3390/ijms21041269.
Cortical spreading depression (CSD) is a propagating wave of depolarization followed by depression of cortical activity. CSD triggers neuroinflammation via the pannexin-1 (Panx1) channel opening, which may eventually cause migraine headaches. However, the regulatory mechanism of Panx1 is unknown. This study investigates whether sarcoma family kinases (SFK) are involved in transmitting CSD-induced Panx1 activation, which is mediated by the NR2A-containing N-methyl-D-aspartate receptor. CSD was induced by topical application of K to cerebral cortices of rats and mouse brain slices. SFK inhibitor, PP2, or NR2A-receptor antagonist, NVP-AAM077, was perfused into contralateral cerebral ventricles (i.c.v.) of rats prior to CSD induction. Co-immunoprecipitation and Western blot were used for detecting protein interactions, and histofluorescence for addressing Panx1 activation. The results demonstrated that PP2 attenuated CSD-induced Panx1 activation in rat ipsilateral cortices. Cortical susceptibility to CSD was reduced by PP2 in rats and by TAT-Panx308 that disrupts SFK-Panx1 interaction in mouse brain slices. Furthermore, CSD promoted activated SFK coupling with Panx1 in rat ipsilateral cortices. Moreover, inhibition of NR2A by NVP-AAM077 reduced elevation of ipsilateral SFK-Panx1 interaction, Panx1 activation induced by CSD and cortical susceptibility to CSD in rats. These data suggest NR2A-regulated, SFK-dependent Panx1 activity plays an important role in migraine aura pathogenesis.
皮层扩散性抑制（CSD）是一种去极化的传播波，随后是皮质活动的抑制。CSD 通过连接蛋白 1（Panx1）通道的开放引发神经炎症，这可能最终导致偏头痛。然而，Panx1 的调节机制尚不清楚。本研究探讨了肉瘤家族激酶（SFK）是否参与传递 CSD 诱导的 Panx1 激活，这种激活是由含有 NR2A 的 N-甲基-D-天冬氨酸受体介导的。CSD 通过在大鼠和小鼠脑片的大脑皮质表面施加 K 来诱导。在诱导 CSD 之前，SFK 抑制剂 PP2 或 NR2A 受体拮抗剂 NVP-AAM077 被灌注到大鼠的对侧侧脑室内（i.c.v.）。共免疫沉淀和 Western blot 用于检测蛋白相互作用，而组织荧光用于解决 Panx1 激活问题。结果表明，PP2 减弱了 CSD 在大鼠同侧皮质中诱导的 Panx1 激活。PP2 降低了大鼠 CSD 诱导的皮层易感性，也降低了 TAT-Panx308 （一种破坏 SFK-Panx1 相互作用的肽）在小鼠脑片中的 CSD 诱导的 Panx1 激活。此外，CSD 促进了激活的 SFK 与 Panx1 在大鼠同侧皮质中的偶联。此外，NVP-AAM077 抑制 NR2A 降低了同侧 SFK-Panx1 相互作用、CSD 诱导的 Panx1 激活和大鼠皮层对 CSD 的易感性的升高。这些数据表明，NR2A 调节的、SFK 依赖性的 Panx1 活性在偏头痛先兆发病机制中起着重要作用。
