目的:探讨酸敏感离子通道1a(ASIC1a)、弱内向整流钾通道相关的酸敏感钾通道1和3(TASK-1和TASK-3)在大鼠癫痫持续状态(SE)中的表达及其在癫痫持续状态中的作用.方法:成年雄性SD大鼠随机分为对照组(control)和SE组,应用氯化锂-匹罗卡品诱...目的:探讨酸敏感离子通道1a(ASIC1a)、弱内向整流钾通道相关的酸敏感钾通道1和3(TASK-1和TASK-3)在大鼠癫痫持续状态(SE)中的表达及其在癫痫持续状态中的作用.方法:成年雄性SD大鼠随机分为对照组(control)和SE组,应用氯化锂-匹罗卡品诱导SE模型,采用real time RT-PCR和Western Blot技术分别检测海马组织ASIC1a、TASK-1和TASK-3在SE后0、1、2和3 h mRNA、蛋白表达水平,应用膜片钳技术观察SE后ASIC1a、TASK-1和TASK-3对海马CA1区锥体神经元兴奋性的影响.结果:在mRNA水平,与control组相比,SE组ASIC1a在2和3h、TASK-1在1h、TASK-3在3h时间点表达显著减少.在蛋白水平,与control组相比,SE组ASIC1a、TASK-1和TASK-3在3h时间点表达均显著降低.与SE control组相比,给予ASIC1a抑制剂后动作电位(AP)的频率明显降低,给予TASK-1和TASK-3抑制剂后,AP的频率均显著增加.结论:SE后大鼠海马区ASIC1a、TASK-1和TASK-3表达下调,且ASIC1a、TASK-1和TASK-3改变了海马CA1区锥体神经元的兴奋性,参与了SE的发生过程.展开更多
Cell replacement therapy using neural progenitor cells(NPCs)has been shown to be an effective treatment for ischemic stroke.However,the therapeutic effect is unsatisfactory due to the imbalanced homeostasis of the loc...Cell replacement therapy using neural progenitor cells(NPCs)has been shown to be an effective treatment for ischemic stroke.However,the therapeutic effect is unsatisfactory due to the imbalanced homeostasis of the local microenvironment after ischemia.Microenvironmental acidosis is a common imbalanced homeostasis in the penumbra and could activate acid-sensing ion channels 1a(ASIC1a),a subunit of proton-gated cation channels following ischemic stroke.However,the role of ASIC1a in NPCs post-ischemia remains elusive.Here,our results indicated that ASIC1a was expressed in NPCs with channel functionality,which could be activated by extracellular acidification.Further evidence revealed that ASIC1a activation inhibited NPC migration and neurogenesis through RhoA signaling-mediated reorganization of filopodia formation,which could be primarily reversed by pharmacological or genetic disruption of ASIC1a.In vivo data showed that the knockout of the ASIC1a gene facilitated NPC migration and neurogenesis in the penumbra to improve behavioral recovery after stroke.展开更多
复发性癫痫诱导慢性树突棘重塑对癫痫发生、终止和长期认知变化很关键,但是调控树突棘重塑的机制并不十分清楚。研究表明,癫痫发作时细胞外[H+]i增加导致组织酸中毒,激活酸敏感离子通道(acid-sensing ion channels,ASICs),引起慢性树突...复发性癫痫诱导慢性树突棘重塑对癫痫发生、终止和长期认知变化很关键,但是调控树突棘重塑的机制并不十分清楚。研究表明,癫痫发作时细胞外[H+]i增加导致组织酸中毒,激活酸敏感离子通道(acid-sensing ion channels,ASICs),引起慢性树突棘重塑。现总结酸中毒和酸敏感离子通道亚型ASIC1a在复发性癫痫引起的树突棘重塑中的作用,重点分析了酸中毒过程的时空变化对痫样放电和树突棘重塑可能的影响,以及酸中毒与ASIC1a在兴奋性和抑制性神经元的功能表达之间的关系,认为ASIC1a可能通过不同机制介导酸中毒在癫痫发生和持续阶段对树突棘的影响。未来研究需要进一步探索癫痫引起的慢性神经元结构和功能改变,阐明酸中毒和ASIC1a在癫痫及其引起的树突棘缺失中的作用。展开更多
Acid-sensing ion channels(ASICs),the main H^(+)receptors in the central nervous system,sense extracellular pH fluctuations and mediate cation influx.ASIC1a,the major subunit responsible for acid-activated current,is w...Acid-sensing ion channels(ASICs),the main H^(+)receptors in the central nervous system,sense extracellular pH fluctuations and mediate cation influx.ASIC1a,the major subunit responsible for acid-activated current,is widely expressed in brain neurons,where it plays pivotal roles in diverse functions including synaptic transmission and plasticity.However,the underlying molecular mechanisms for these functions remain mysterious.Using extracellular epitope tagging and a novel antibody recognizing the hASIC1a ectodomain,we examined the membrane targeting and dynamic trafficking of hASIC1a in cultured cortical neurons.Surface hASIC1a was distributed throughout somata and dendrites,clustered in spine heads,and co-localized with postsynaptic markers.By extracellular pHluorin tagging and fluorescence recovery after photobleaching,we detected movement of hASIC1a in synaptic spine heads.Single-particle tracking along with use of the anti-hASIC1a ectodomain antibody revealed long-distance migration and local movement of surface hASIC1a puncta on dendrites.Importantly,enhancing synaptic activity with brain-derived neurotrophic factor accelerated the trafficking and lateral mobility of hASIC1a.With this newly-developed toolbox,our data demonstrate the synaptic location and high dynamics of functionallyrelevant hASIC1a on the surface of excitatory synapses,supporting its involvement in synaptic functions.展开更多
目的探讨内皮祖细胞(EPCs)移植对百草枯(PQ)中毒所致的急性肺损伤(ALI)小鼠的作用机制。方法采用密度梯度离心联合贴壁培养法分离培养小鼠外周血EPC,为后期移植提供供体细胞。应用腹腔注射法建立百草枯所致的急性肺损伤(PQ-ALI)小鼠模型...目的探讨内皮祖细胞(EPCs)移植对百草枯(PQ)中毒所致的急性肺损伤(ALI)小鼠的作用机制。方法采用密度梯度离心联合贴壁培养法分离培养小鼠外周血EPC,为后期移植提供供体细胞。应用腹腔注射法建立百草枯所致的急性肺损伤(PQ-ALI)小鼠模型,建模成功后小鼠分为四组:正常对照组、模型组、PQ损伤组、EPCs治疗组(每组9只),移植后48 h处死小鼠并取肺组织,检测肺组织湿干质量(W/D)比值、肿瘤坏死因子-a(TNF-a)含量及肺组织中酸敏感离子通道-l a(ASICla)的表达。结果①正常对照组与模型组比较小鼠肺W/D无明显变化(两者W/D比值分别为3.72、3.67),差异无统计学意义(P>0.05);PQ损伤组小鼠肺组织W/D比值为9.80,与对照组比较差异有统计学意义(P<0.05);尾静脉E PC移植后(EPCs组)可以明显降低小鼠肺组织W/D比值,其值为4.91,与P Q损伤组比较差异有统计学意义(P<0.05)。②正常对照组及模型组中TNF-a分别为226 n g/L和204 n g/L,两组比较差异无统计学意义(P>0.05);PQ损伤组小鼠肺组织中TNF-a为382 ng/L明显增高,与正常对照组比较差异有统计学意义(P<0.05);EPCs治疗组小鼠肺组织TNF-a为276 n g/L呈下降趋势,与P Q损伤组比较差异有统计学意义(P<0.05)。③PQ损伤组和EPCs治疗组小鼠肺组织中ASICU的表达量分别为1.73和1.12,均较正常对照组ASICla的表达量(0.56)明显增多,差异有统计学意义(P<0.05);EPCs治疗组小鼠肺组织中ASICla的表达量低于PQ损伤组(P<0.05);模型组小鼠ASICla的表达量为0.54和治疗对照组小鼠肺组织中ASICla的表达量最少,两组比较差异无统计学意义(P>0.05)。结论PQ-ALI小鼠模型机制可能和ASICla激活有关,EPC移植可改善PQ中毒所致ALI小鼠状态,可能和下调ASIC1a的表达相关,但具体机制尚未清楚。展开更多
酸敏感离子通道(acid-sensing ion channels,ASICs)是细胞外酸性p H值的关键受体,在生物体内广泛表达,在疼痛、炎症、记忆、脑缺血等多项涉及酸中毒的病理和生理过程中起着重要作用。它能介导肿瘤细胞的迁移和侵袭,参与肿瘤的发展。ASIC...酸敏感离子通道(acid-sensing ion channels,ASICs)是细胞外酸性p H值的关键受体,在生物体内广泛表达,在疼痛、炎症、记忆、脑缺血等多项涉及酸中毒的病理和生理过程中起着重要作用。它能介导肿瘤细胞的迁移和侵袭,参与肿瘤的发展。ASIC1在肺癌、胰腺癌、乳腺癌等实体肿瘤中大量表达,其表达对肿瘤早期诊断及治疗和预后起到重要的作用。目前关于ASIC1在肿瘤中的具体调控机制仍不是十分清楚。展开更多
文摘目的:探讨酸敏感离子通道1a(ASIC1a)、弱内向整流钾通道相关的酸敏感钾通道1和3(TASK-1和TASK-3)在大鼠癫痫持续状态(SE)中的表达及其在癫痫持续状态中的作用.方法:成年雄性SD大鼠随机分为对照组(control)和SE组,应用氯化锂-匹罗卡品诱导SE模型,采用real time RT-PCR和Western Blot技术分别检测海马组织ASIC1a、TASK-1和TASK-3在SE后0、1、2和3 h mRNA、蛋白表达水平,应用膜片钳技术观察SE后ASIC1a、TASK-1和TASK-3对海马CA1区锥体神经元兴奋性的影响.结果:在mRNA水平,与control组相比,SE组ASIC1a在2和3h、TASK-1在1h、TASK-3在3h时间点表达显著减少.在蛋白水平,与control组相比,SE组ASIC1a、TASK-1和TASK-3在3h时间点表达均显著降低.与SE control组相比,给予ASIC1a抑制剂后动作电位(AP)的频率明显降低,给予TASK-1和TASK-3抑制剂后,AP的频率均显著增加.结论:SE后大鼠海马区ASIC1a、TASK-1和TASK-3表达下调,且ASIC1a、TASK-1和TASK-3改变了海马CA1区锥体神经元的兴奋性,参与了SE的发生过程.
基金the National Natural Science Foundation of China(81873771,81371340,and 82271424)the Key Project of Natural Science Foundation of Chongqing(cstc2013jjB012503).
文摘Cell replacement therapy using neural progenitor cells(NPCs)has been shown to be an effective treatment for ischemic stroke.However,the therapeutic effect is unsatisfactory due to the imbalanced homeostasis of the local microenvironment after ischemia.Microenvironmental acidosis is a common imbalanced homeostasis in the penumbra and could activate acid-sensing ion channels 1a(ASIC1a),a subunit of proton-gated cation channels following ischemic stroke.However,the role of ASIC1a in NPCs post-ischemia remains elusive.Here,our results indicated that ASIC1a was expressed in NPCs with channel functionality,which could be activated by extracellular acidification.Further evidence revealed that ASIC1a activation inhibited NPC migration and neurogenesis through RhoA signaling-mediated reorganization of filopodia formation,which could be primarily reversed by pharmacological or genetic disruption of ASIC1a.In vivo data showed that the knockout of the ASIC1a gene facilitated NPC migration and neurogenesis in the penumbra to improve behavioral recovery after stroke.
文摘复发性癫痫诱导慢性树突棘重塑对癫痫发生、终止和长期认知变化很关键,但是调控树突棘重塑的机制并不十分清楚。研究表明,癫痫发作时细胞外[H+]i增加导致组织酸中毒,激活酸敏感离子通道(acid-sensing ion channels,ASICs),引起慢性树突棘重塑。现总结酸中毒和酸敏感离子通道亚型ASIC1a在复发性癫痫引起的树突棘重塑中的作用,重点分析了酸中毒过程的时空变化对痫样放电和树突棘重塑可能的影响,以及酸中毒与ASIC1a在兴奋性和抑制性神经元的功能表达之间的关系,认为ASIC1a可能通过不同机制介导酸中毒在癫痫发生和持续阶段对树突棘的影响。未来研究需要进一步探索癫痫引起的慢性神经元结构和功能改变,阐明酸中毒和ASIC1a在癫痫及其引起的树突棘缺失中的作用。
基金This work was supported by grants from the National Natural Science Foundation of China(81961128024 and 81730095)the Innovative Research Team of High-level Local Universities in Shanghai+2 种基金the Science and Technology Commission of Shanghai Municipality(18JC1420302)the Shanghai Municipal Science and Technology Major Project(2018SHZDZX05)the US National Institutes of Health(NS114716).
文摘Acid-sensing ion channels(ASICs),the main H^(+)receptors in the central nervous system,sense extracellular pH fluctuations and mediate cation influx.ASIC1a,the major subunit responsible for acid-activated current,is widely expressed in brain neurons,where it plays pivotal roles in diverse functions including synaptic transmission and plasticity.However,the underlying molecular mechanisms for these functions remain mysterious.Using extracellular epitope tagging and a novel antibody recognizing the hASIC1a ectodomain,we examined the membrane targeting and dynamic trafficking of hASIC1a in cultured cortical neurons.Surface hASIC1a was distributed throughout somata and dendrites,clustered in spine heads,and co-localized with postsynaptic markers.By extracellular pHluorin tagging and fluorescence recovery after photobleaching,we detected movement of hASIC1a in synaptic spine heads.Single-particle tracking along with use of the anti-hASIC1a ectodomain antibody revealed long-distance migration and local movement of surface hASIC1a puncta on dendrites.Importantly,enhancing synaptic activity with brain-derived neurotrophic factor accelerated the trafficking and lateral mobility of hASIC1a.With this newly-developed toolbox,our data demonstrate the synaptic location and high dynamics of functionallyrelevant hASIC1a on the surface of excitatory synapses,supporting its involvement in synaptic functions.
文摘目的探讨内皮祖细胞(EPCs)移植对百草枯(PQ)中毒所致的急性肺损伤(ALI)小鼠的作用机制。方法采用密度梯度离心联合贴壁培养法分离培养小鼠外周血EPC,为后期移植提供供体细胞。应用腹腔注射法建立百草枯所致的急性肺损伤(PQ-ALI)小鼠模型,建模成功后小鼠分为四组:正常对照组、模型组、PQ损伤组、EPCs治疗组(每组9只),移植后48 h处死小鼠并取肺组织,检测肺组织湿干质量(W/D)比值、肿瘤坏死因子-a(TNF-a)含量及肺组织中酸敏感离子通道-l a(ASICla)的表达。结果①正常对照组与模型组比较小鼠肺W/D无明显变化(两者W/D比值分别为3.72、3.67),差异无统计学意义(P>0.05);PQ损伤组小鼠肺组织W/D比值为9.80,与对照组比较差异有统计学意义(P<0.05);尾静脉E PC移植后(EPCs组)可以明显降低小鼠肺组织W/D比值,其值为4.91,与P Q损伤组比较差异有统计学意义(P<0.05)。②正常对照组及模型组中TNF-a分别为226 n g/L和204 n g/L,两组比较差异无统计学意义(P>0.05);PQ损伤组小鼠肺组织中TNF-a为382 ng/L明显增高,与正常对照组比较差异有统计学意义(P<0.05);EPCs治疗组小鼠肺组织TNF-a为276 n g/L呈下降趋势,与P Q损伤组比较差异有统计学意义(P<0.05)。③PQ损伤组和EPCs治疗组小鼠肺组织中ASICU的表达量分别为1.73和1.12,均较正常对照组ASICla的表达量(0.56)明显增多,差异有统计学意义(P<0.05);EPCs治疗组小鼠肺组织中ASICla的表达量低于PQ损伤组(P<0.05);模型组小鼠ASICla的表达量为0.54和治疗对照组小鼠肺组织中ASICla的表达量最少,两组比较差异无统计学意义(P>0.05)。结论PQ-ALI小鼠模型机制可能和ASICla激活有关,EPC移植可改善PQ中毒所致ALI小鼠状态,可能和下调ASIC1a的表达相关,但具体机制尚未清楚。
文摘酸敏感离子通道(acid-sensing ion channels,ASICs)是细胞外酸性p H值的关键受体,在生物体内广泛表达,在疼痛、炎症、记忆、脑缺血等多项涉及酸中毒的病理和生理过程中起着重要作用。它能介导肿瘤细胞的迁移和侵袭,参与肿瘤的发展。ASIC1在肺癌、胰腺癌、乳腺癌等实体肿瘤中大量表达,其表达对肿瘤早期诊断及治疗和预后起到重要的作用。目前关于ASIC1在肿瘤中的具体调控机制仍不是十分清楚。
