Background Tactile and mechanical pain are crucial to our interaction with the environment,yet the underpinning molecular mechanism is still elusive.Endophilin A2(EndoA2)is an evolutionarily conserved protein that is ...Background Tactile and mechanical pain are crucial to our interaction with the environment,yet the underpinning molecular mechanism is still elusive.Endophilin A2(EndoA2)is an evolutionarily conserved protein that is documented in the endocytosis pathway.However,the role of EndoA2 in the regulation of mechanical sensitivity and its underlying mechanisms are currently unclear.Methods Male and female C57BL/6 mice(8–12 weeks)and male cynomolgus monkeys(7–10 years old)were used in our experiments.Nerve injury-,inflammatory-,and chemotherapy-induced pathological pain models were established for this study.Behavioral tests of touch,mechanical pain,heat pain,and cold pain were performed in mice and nonhuman primates.Western blotting,immunostaining,co-immunoprecipitation,proximity ligation and patch-clamp recordings were performed to gain insight into the mechanisms.Results The results showed that EndoA2 was primarily distributed in neurofilament-200-positive(NF200+)medium-to-large diameter dorsal root ganglion(DRG)neurons of mice and humans.Loss of EndoA2 in mouse NF200+DRG neurons selectively impaired the tactile and mechanical allodynia.Furthermore,EndoA2 interacted with the mechanically sensitive ion channel Piezo2 and promoted the membrane trafficking of Piezo2 in DRG neurons.Moreover,as an adaptor protein,EndoA2 also bound to kinesin family member 5B(KIF5B),which was involved in the EndoA2-mediated membrane trafficking process of Piezo2.Loss of EndoA2 in mouse DRG neurons damaged Piezo2-mediated rapidly adapting mechanically activated currents,and re-expression of EndoA2 rescued the MA currents.In addition,interference with EndoA2 also suppressed touch sensitivity and mechanical hypersensitivity in nonhuman primates.Conclusions Our data reveal that the KIF5B/EndoA2/Piezo2 complex is essential for Piezo2 trafficking and for sustaining transmission of touch and mechanical hypersensitivity signals.EndoA2 regulates touch and mechanical allodynia via kinesin-mediated Piezo2 trafficking in sensory neurons.Our findings identify a potential new target for the treatment of mechanical pain.展开更多
目的探讨压电型机械敏感离子通道元件2(piezoelectric type mechanosensitive ion channel component 2,PIEZO2)对胰腺癌细胞发生发展的影响。方法采用免疫组织化学技术检测胰腺癌与癌旁组织的PIEZO2阳性率;将PIEZO2干扰片段转染到胰腺...目的探讨压电型机械敏感离子通道元件2(piezoelectric type mechanosensitive ion channel component 2,PIEZO2)对胰腺癌细胞发生发展的影响。方法采用免疫组织化学技术检测胰腺癌与癌旁组织的PIEZO2阳性率;将PIEZO2干扰片段转染到胰腺癌细胞Panc-1中,通过实时荧光定量PCR(qPCR)和蛋白免疫印迹(WB)检测PIEZO2的转录水平及蛋白水平。以细胞计数试剂盒-8(Cell Counting Kit-8,CCK8)检测细胞增殖情况,transwell法检测细胞的迁移侵袭能力。通过Illumina HiSeq^(TM)X Ten进行cDNA文库构建并测序,分析干扰PIEZO2后Panc-1细胞内差异基因并通过GO富集分析参与基因富集的相关通路。结果胰腺癌组织中PIEZO2表达水平高于癌旁组织,且高PIEZO2表达的胰腺癌患者总生存率较低。敲减PIEZO2基因后,qPCR和WB结果显示PIEZO2的表达量较对照组减低,差异具有统计学意义。敲减组的增殖、迁移、侵袭能力均低于对照组,且差异具有统计学意义。对两组进行转录组分析发现,两组分别获得15521,15325个基因,与敲减组相比,对照组有3411个差异表达基因(differentially expressed genes,DEGs)上调,2830个DEGs下调,GO富集分析发现DEGs基因主要富集于细胞内解剖结构、细胞连接和细胞外基质。结论敲除PIEZO2可抑制细胞的增殖、迁移和侵袭,PIEZO2通过调控细胞解剖结构、细胞连接和细胞外基质参与胰腺癌的发生发展,靶向PIEZO2将是胰腺癌新的治疗策略。展开更多
Almost every life form,from the tiniest bacterium to humans,is mechanosensitive,implying it can use mechani-cal stresses to trigger certain physiological responses in the form of electric signals.Mechanotransduction l...Almost every life form,from the tiniest bacterium to humans,is mechanosensitive,implying it can use mechani-cal stresses to trigger certain physiological responses in the form of electric signals.Mechanotransduction largely relies on ion channels that respond to mechanical forces,such as the epithelial sodium channels/degenerins,tran-sient receptor potential channel,and the two-pore domain potassium channel.Piezo1 and Piezo2 proteins were discovered to be the biggest non-selective mechanosensitive cation channels in the cell membrane.A substantial amount of research has previously been published on the Piezo channel’s function in touch sensation,balance,and cardiovascular regression.However,the mechanistic perspective must be refined to fully understand the role of Piezo proteins in tissue engineering.This review centers on the latest insights into the structure of Piezo chan-nels,activation mechanisms,and its interactions with cytoskeletal components,by emphasizing the physiological activities of Piezo channels in different tissues.The study also places focus on the possibilities of targeting this cation channel family as a tissue regeneration aid.展开更多
基金National Natural Science Foundation of China(82271241 and 82001172 to XLZ,81801112 to MXX)Guangdong Basic and Applied Basic Research Foundation(2022A1515012389 to MXX,2022A1515012543 to RCL)+2 种基金Young Talent Support Project of Guangzhou Association for Science and Technology(QT20220101169 to XLZ)Excellent Young Talents Project of Guangdong Provincial People’s Hospital,Guangdong Academy of Medical Sciences(KY012021188 to XLZ)Science and Technology Projects in Guangzhou(202201010792 to RCL)。
文摘Background Tactile and mechanical pain are crucial to our interaction with the environment,yet the underpinning molecular mechanism is still elusive.Endophilin A2(EndoA2)is an evolutionarily conserved protein that is documented in the endocytosis pathway.However,the role of EndoA2 in the regulation of mechanical sensitivity and its underlying mechanisms are currently unclear.Methods Male and female C57BL/6 mice(8–12 weeks)and male cynomolgus monkeys(7–10 years old)were used in our experiments.Nerve injury-,inflammatory-,and chemotherapy-induced pathological pain models were established for this study.Behavioral tests of touch,mechanical pain,heat pain,and cold pain were performed in mice and nonhuman primates.Western blotting,immunostaining,co-immunoprecipitation,proximity ligation and patch-clamp recordings were performed to gain insight into the mechanisms.Results The results showed that EndoA2 was primarily distributed in neurofilament-200-positive(NF200+)medium-to-large diameter dorsal root ganglion(DRG)neurons of mice and humans.Loss of EndoA2 in mouse NF200+DRG neurons selectively impaired the tactile and mechanical allodynia.Furthermore,EndoA2 interacted with the mechanically sensitive ion channel Piezo2 and promoted the membrane trafficking of Piezo2 in DRG neurons.Moreover,as an adaptor protein,EndoA2 also bound to kinesin family member 5B(KIF5B),which was involved in the EndoA2-mediated membrane trafficking process of Piezo2.Loss of EndoA2 in mouse DRG neurons damaged Piezo2-mediated rapidly adapting mechanically activated currents,and re-expression of EndoA2 rescued the MA currents.In addition,interference with EndoA2 also suppressed touch sensitivity and mechanical hypersensitivity in nonhuman primates.Conclusions Our data reveal that the KIF5B/EndoA2/Piezo2 complex is essential for Piezo2 trafficking and for sustaining transmission of touch and mechanical hypersensitivity signals.EndoA2 regulates touch and mechanical allodynia via kinesin-mediated Piezo2 trafficking in sensory neurons.Our findings identify a potential new target for the treatment of mechanical pain.
文摘Almost every life form,from the tiniest bacterium to humans,is mechanosensitive,implying it can use mechani-cal stresses to trigger certain physiological responses in the form of electric signals.Mechanotransduction largely relies on ion channels that respond to mechanical forces,such as the epithelial sodium channels/degenerins,tran-sient receptor potential channel,and the two-pore domain potassium channel.Piezo1 and Piezo2 proteins were discovered to be the biggest non-selective mechanosensitive cation channels in the cell membrane.A substantial amount of research has previously been published on the Piezo channel’s function in touch sensation,balance,and cardiovascular regression.However,the mechanistic perspective must be refined to fully understand the role of Piezo proteins in tissue engineering.This review centers on the latest insights into the structure of Piezo chan-nels,activation mechanisms,and its interactions with cytoskeletal components,by emphasizing the physiological activities of Piezo channels in different tissues.The study also places focus on the possibilities of targeting this cation channel family as a tissue regeneration aid.
