OBJECTIVE The high prevalence of sleep disturbance has been found in patients with striatum-related neurodegenerative disorders.In the striatum,there are abundant adenosine A2A receptors(A2ARs)whichhavebeen reported t...OBJECTIVE The high prevalence of sleep disturbance has been found in patients with striatum-related neurodegenerative disorders.In the striatum,there are abundant adenosine A2A receptors(A2ARs)whichhavebeen reported to mediatesleepbehavior for adenosine.We hypothesized that the A2AR-expressing neurons in the striatum are involved in sleep-wake regulation.METHODS We employed a chemogenetic technique,designer receptor exclusively activated by designer drug(DREADD),to specifically and non-invasively manipulate the neuron activity based on the principle of Cre/Lox P recombination,EEG/electromyogram recording for sleep-wake behaviors,the neural tracing approach toselectively visualize the perikarya of A2AR-expressing neurons and their axons by adeno-associated virus(AAV)encoding humanized Renilla green fluorescent(hr GFP)as a tracerin A2AR-Cre mice.In addition,we used immunoelectron microscopy,patch-clamp technique,and optogenetics in A2AR-Cre mice to selectively characterize the synapse and functional connectivity between the A2AR-expressing neurons and the neuron of their downstream targets in vitro.RESULTS The activation of A2AR-expressing neurons in rostral,centromedial and centrolateral striatum increased non-rapid eye movement(non-REM,NREM)sleep,concomitant with a reduction in wakefulness,whereas the activation of A2AR-expressing neurons in caudal striatum didn′t alter sleep-wake profiles at all.Topographical projections in the sagittal section showed that the axons of A2ARexpressing neurons from rostral striatum distributed in the rostral external globuspallidus(GPe)with a discoidal region paralleled to the striato-pallidal border,while the axons of the A2AR-expressing neurons from the central striatum not only distributed in the rostral GPe,but also in the caudal GPe with a similar distributing pattern as did in rostral neurons.However,the axons of A2ARexpressing neurons from caudal striatum just scattered in the caudal GPe.Based on our anatomical findings and patch-clamp technique combining with optogenetics,we found that A2AR neurons in the rostral striatum preferentially formed inhibitory synapses with parvalbumin(PV)-positive neurons in the rostral GPe,while A2AR neurons in the caudal striatum preferentially formed inhibitory synapses with PV-negative neurons in the caudal GPe.CONCLUSION The present results indicated that the A2AR-expressing neurons in rostral and central striatum are involved in sleep-wake regulation,probably via innervating PV-positive neurons in the GPe.展开更多
目的通过检测降钙素基因相关肽(calcitonin gene related peptide,CGRP)及腺苷2A受体(adenosine 2Areceptor,A2AR)蛋白在偏头痛模型大鼠三叉神经组织中的表达,探讨CGRP和A2AR在大鼠偏头痛发病机制中的作用。方法将30只雄性Sprague-Dawle...目的通过检测降钙素基因相关肽(calcitonin gene related peptide,CGRP)及腺苷2A受体(adenosine 2Areceptor,A2AR)蛋白在偏头痛模型大鼠三叉神经组织中的表达,探讨CGRP和A2AR在大鼠偏头痛发病机制中的作用。方法将30只雄性Sprague-Dawley(SD)大鼠随机分为对照组和模型组,每组15只。模型组采用每周1次皮下注射硝酸甘油法建立无先兆性偏头痛模型,共5周;对照组则用生理盐水皮下注射代替硝酸甘油。对两组大鼠进行行为学评分,而后采集两组大鼠颅底三叉神经节,采用免疫组化法检测三叉神经节中CGRP的表达;采用免疫荧光检测法观察三叉神经节中A2AR表达;采用westernblot法检测A2AR蛋白表达量。结果三叉神经节组织HE染色结果:模型组和对照组均显示神经元与神经纤维分布正常、无紊乱,解剖结构完整,无异常改变。三叉神经节中A2AR蛋白表达的Western-blot量及灰度值测定结果:模型组A2AR蛋白表达量及灰度值高于对照组。差异均有统计学意义(P<0.01)。三叉神经组织中A2AR表达蛋白免疫荧光测定结果及A2AR阳性神经元细胞数:模型组中A2AR在三叉神经各神经元中的免疫荧光的表达分布明显多于对照组模型组。模型组表达A2AR的阳性神经元细胞数高于对照组,差异有统计学意义(P<0.01)。三叉神经节CGRP免疫组化表达水平及OD值结果:模型组CGRP及OD值的表达水平明显高于对照组,差异有统计学意义(P<0.01)。结论模型大鼠偏头痛发作时,三叉神经组织无解剖结构上的改变,大鼠行为学改变与三叉神经功能异常有关。A2AR的激活或蛋白上调,CGRP的大量释放。A2AR、CGRP可能在偏头痛的发作中起重要作用。展开更多
Hepatic ischemia–reperfusion injury(HIRI)is an important cause of liver injury following liver transplantation and major resections,and neutrophils are the key effector cells in HIRI.Double-negative T regulatory cell...Hepatic ischemia–reperfusion injury(HIRI)is an important cause of liver injury following liver transplantation and major resections,and neutrophils are the key effector cells in HIRI.Double-negative T regulatory cells(DNT)are increasingly recognized as having critical regulatory functions in the immune system.Whether DNT expresses distinct immunoregulatory mechanisms to modulate neutrophils,as in HIRI,remains largely unknown.In this study,we found that murine and human DNT highly expressed CD39that protected DNT from extracellular ATP-induced apoptosis and generated adenosine in tandem with CD73,to induce high levels of neutrophil apoptosis.Furthermore,extracellular adenosine enhanced DNT survival and suppressive function by upregulating survivin and NKG2D expression via the A2AR/pAKT/FOXO1 signaling pathway.Adoptive transfer of DNT ameliorated HIRI in mice through the inhibition of neutrophils in a CD39-dependent manner.Lastly,the adoptive transfer of A2ar^(-/-)DNT validated the importance of adenosine/A2AR signaling,in promoting DNT survival and immunomodulatory function to protect against HIRI in vivo.In conclusion,purinergic signaling is crucial for DNT homeostasis in HIRI.Augmentation of CD39 or activation of A2AR signaling in DNT may provide novel therapeutic strategies to target innate immune disorders.展开更多
Tumors survive by creating a tumor microenvironment(TME)that suppresses antitumor immunity.The TME suppresses the immune system by limiting antigen presentation,inhibiting lymphocyte and natural killer(NK)cell activat...Tumors survive by creating a tumor microenvironment(TME)that suppresses antitumor immunity.The TME suppresses the immune system by limiting antigen presentation,inhibiting lymphocyte and natural killer(NK)cell activation,and facilitating T cell exhaustion.Checkpoint inhibitors like anti-PD-1 and anti-CTLA4 are immunostimulatory antibodies,and their blockade extends the survival of some but not all cancer patients.Extracellular adenosine triphosphate(ATP)is abundant in inflamed tumors,and its metabolite,adenosine(ADO),is a driver of immunosuppression mediated by adenosine A2A receptors(A2AR)and adenosine A2B receptors(A2BR)found on tumor-associated lymphoid and myeloid cells.This review will focus on adenosine as a key checkpoint inhibitor-like immunosuppressive player in the TME and how reducing adenosine production or blocking A2AR and A2BR enhances antitumor immunity.展开更多
基金The project supported by National Basic Research Program of China(2015CB856401)
文摘OBJECTIVE The high prevalence of sleep disturbance has been found in patients with striatum-related neurodegenerative disorders.In the striatum,there are abundant adenosine A2A receptors(A2ARs)whichhavebeen reported to mediatesleepbehavior for adenosine.We hypothesized that the A2AR-expressing neurons in the striatum are involved in sleep-wake regulation.METHODS We employed a chemogenetic technique,designer receptor exclusively activated by designer drug(DREADD),to specifically and non-invasively manipulate the neuron activity based on the principle of Cre/Lox P recombination,EEG/electromyogram recording for sleep-wake behaviors,the neural tracing approach toselectively visualize the perikarya of A2AR-expressing neurons and their axons by adeno-associated virus(AAV)encoding humanized Renilla green fluorescent(hr GFP)as a tracerin A2AR-Cre mice.In addition,we used immunoelectron microscopy,patch-clamp technique,and optogenetics in A2AR-Cre mice to selectively characterize the synapse and functional connectivity between the A2AR-expressing neurons and the neuron of their downstream targets in vitro.RESULTS The activation of A2AR-expressing neurons in rostral,centromedial and centrolateral striatum increased non-rapid eye movement(non-REM,NREM)sleep,concomitant with a reduction in wakefulness,whereas the activation of A2AR-expressing neurons in caudal striatum didn′t alter sleep-wake profiles at all.Topographical projections in the sagittal section showed that the axons of A2ARexpressing neurons from rostral striatum distributed in the rostral external globuspallidus(GPe)with a discoidal region paralleled to the striato-pallidal border,while the axons of the A2AR-expressing neurons from the central striatum not only distributed in the rostral GPe,but also in the caudal GPe with a similar distributing pattern as did in rostral neurons.However,the axons of A2ARexpressing neurons from caudal striatum just scattered in the caudal GPe.Based on our anatomical findings and patch-clamp technique combining with optogenetics,we found that A2AR neurons in the rostral striatum preferentially formed inhibitory synapses with parvalbumin(PV)-positive neurons in the rostral GPe,while A2AR neurons in the caudal striatum preferentially formed inhibitory synapses with PV-negative neurons in the caudal GPe.CONCLUSION The present results indicated that the A2AR-expressing neurons in rostral and central striatum are involved in sleep-wake regulation,probably via innervating PV-positive neurons in the GPe.
文摘目的通过检测降钙素基因相关肽(calcitonin gene related peptide,CGRP)及腺苷2A受体(adenosine 2Areceptor,A2AR)蛋白在偏头痛模型大鼠三叉神经组织中的表达,探讨CGRP和A2AR在大鼠偏头痛发病机制中的作用。方法将30只雄性Sprague-Dawley(SD)大鼠随机分为对照组和模型组,每组15只。模型组采用每周1次皮下注射硝酸甘油法建立无先兆性偏头痛模型,共5周;对照组则用生理盐水皮下注射代替硝酸甘油。对两组大鼠进行行为学评分,而后采集两组大鼠颅底三叉神经节,采用免疫组化法检测三叉神经节中CGRP的表达;采用免疫荧光检测法观察三叉神经节中A2AR表达;采用westernblot法检测A2AR蛋白表达量。结果三叉神经节组织HE染色结果:模型组和对照组均显示神经元与神经纤维分布正常、无紊乱,解剖结构完整,无异常改变。三叉神经节中A2AR蛋白表达的Western-blot量及灰度值测定结果:模型组A2AR蛋白表达量及灰度值高于对照组。差异均有统计学意义(P<0.01)。三叉神经组织中A2AR表达蛋白免疫荧光测定结果及A2AR阳性神经元细胞数:模型组中A2AR在三叉神经各神经元中的免疫荧光的表达分布明显多于对照组模型组。模型组表达A2AR的阳性神经元细胞数高于对照组,差异有统计学意义(P<0.01)。三叉神经节CGRP免疫组化表达水平及OD值结果:模型组CGRP及OD值的表达水平明显高于对照组,差异有统计学意义(P<0.01)。结论模型大鼠偏头痛发作时,三叉神经组织无解剖结构上的改变,大鼠行为学改变与三叉神经功能异常有关。A2AR的激活或蛋白上调,CGRP的大量释放。A2AR、CGRP可能在偏头痛的发作中起重要作用。
基金supported by the grants from the National Natural Science Foundation of China(81970503,82100670,82202021and 82270606)Chinese Institutes for Medical Research+7 种基金Beijing(CX24PY16)R&D Program of Beijing Municipal Education Commission(KZ202210025036)Beijing Municipal Administration of Hospitals’Ascent Plan(DFL20220103)Beijing Nova Program(Z211100002121036)Youth Beijing Scholar(035)and the Reform and Development Program of Beijing Institute of Respiratory Medicine(Ggyfz202403)Generation of reagents and mutant mice are also supported by the National Institutes of Health(R01 DK108894R21 CA164970 and R21 CA221702)Department of Defense Award W81XWH-16-0464。
文摘Hepatic ischemia–reperfusion injury(HIRI)is an important cause of liver injury following liver transplantation and major resections,and neutrophils are the key effector cells in HIRI.Double-negative T regulatory cells(DNT)are increasingly recognized as having critical regulatory functions in the immune system.Whether DNT expresses distinct immunoregulatory mechanisms to modulate neutrophils,as in HIRI,remains largely unknown.In this study,we found that murine and human DNT highly expressed CD39that protected DNT from extracellular ATP-induced apoptosis and generated adenosine in tandem with CD73,to induce high levels of neutrophil apoptosis.Furthermore,extracellular adenosine enhanced DNT survival and suppressive function by upregulating survivin and NKG2D expression via the A2AR/pAKT/FOXO1 signaling pathway.Adoptive transfer of DNT ameliorated HIRI in mice through the inhibition of neutrophils in a CD39-dependent manner.Lastly,the adoptive transfer of A2ar^(-/-)DNT validated the importance of adenosine/A2AR signaling,in promoting DNT survival and immunomodulatory function to protect against HIRI in vivo.In conclusion,purinergic signaling is crucial for DNT homeostasis in HIRI.Augmentation of CD39 or activation of A2AR signaling in DNT may provide novel therapeutic strategies to target innate immune disorders.
文摘Tumors survive by creating a tumor microenvironment(TME)that suppresses antitumor immunity.The TME suppresses the immune system by limiting antigen presentation,inhibiting lymphocyte and natural killer(NK)cell activation,and facilitating T cell exhaustion.Checkpoint inhibitors like anti-PD-1 and anti-CTLA4 are immunostimulatory antibodies,and their blockade extends the survival of some but not all cancer patients.Extracellular adenosine triphosphate(ATP)is abundant in inflamed tumors,and its metabolite,adenosine(ADO),is a driver of immunosuppression mediated by adenosine A2A receptors(A2AR)and adenosine A2B receptors(A2BR)found on tumor-associated lymphoid and myeloid cells.This review will focus on adenosine as a key checkpoint inhibitor-like immunosuppressive player in the TME and how reducing adenosine production or blocking A2AR and A2BR enhances antitumor immunity.
