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
The Chinese herbs Herba Epimedii, Fructus Ligustri Lucidi and Rhizoma Polygonati were injected into Parkinson＇s disease mice established via intraperitoneal injection of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridin...The Chinese herbs Herba Epimedii, Fructus Ligustri Lucidi and Rhizoma Polygonati were injected into Parkinson＇s disease mice established via intraperitoneal injection of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine hydrochloride. The selective monoamine oxidase B inhibitor selegiline was used as a positive control drug. After successive administration for 4 weeks, Herba Epimedii could downregulate the expression of caspase-3 and increase the brain-derived neurotrophic factor level, as well as increase tyrosine hydroxylase activity in the substantia nigra of Parkinson＇s disease mouse models. Rhizoma Polygonaticould downregulate the expression of caspase-3 and FasL, and increase neural growth factor and brain-derived neurotrophic factor levels. Fructus Ligustn Lucidi could downregulate caspase-3 expression. Rhizoma Polygonati and Fructus Ligustn Lucidi did not produce obvious effects on tyrosine hydroxylase activity. Herba Epimedii and Fructus Ligustri Lucidi yielded similar effects on apoptosis-promoting factors to those elicited by selegiline. Herba Epimedii and Rhizoma Polygonati significantly increased the levels of neurotrophic factors compared with selegiline. Herba Epimedii significantly increased tyrosine hydroxylase activity compared with selegiline. It is indicated that the kidney-tonifying Chinese herbal preparation can downregulate the expression of apoptosis-promoting factors, increase neurotrophic factors levels in the substantia nigra and striatum, as well as increase tyrosine hydroxylase activity in the substantia nigra of Parkinson＇s disease mouse models, thereby exerting a stronger or similar neuroprotective effects compared with selegiline.展开更多
The over-expression of α-synuclein is a major factor in the death of dopaminergic neurons in a methamphetamine-induced model of Parkinson’s disease. In the present study, α-synuclein knockdown rats were created by ...The over-expression of α-synuclein is a major factor in the death of dopaminergic neurons in a methamphetamine-induced model of Parkinson’s disease. In the present study, α-synuclein knockdown rats were created by injecting α-synuclein-shRNA lentivirus stereotaxically into the right striatum of experimental rats. At 2 weeks post-injection, the rats were injected intraper-itoneally with methamphetamine to establish the model of Parkinson’s disease. Expression of α-synuclein mRNA and protein in the right striatum of the injected rats was significantly down-regulated. Food intake and body weight were greater in α-synuclein knockdown rats, and water intake and stereotyped behavior score were lower than in model rats. Striatal dopamine and tyrosine hydroxylase levels were significantly elevated in α-synuclein knockdown rats. Moreover, superoxide dismutase activity was greater in α-synuclein knockdown rat striatum, but the levels of reactive oxygen species, malondialdehyde, nitric oxide synthase and nitrogen monoxide were lower compared with model rats. We also found that α-synuclein knockdown inhibited metham-phetamine-induced neuronal apoptosis. These results suggest that α-synuclein has the capacity to reverse methamphetamine-induced apoptosis of dopaminergic neurons in the rat striatum by inhibiting oxidative stress and improving dopaminergic system function.展开更多
A new technology called in vivo glia-to-neuron conversion has emerged in recent years as a promising next generation therapy for neural regeneration and repair. This is achieved through reprogramming endogenous glial ...A new technology called in vivo glia-to-neuron conversion has emerged in recent years as a promising next generation therapy for neural regeneration and repair. This is achieved through reprogramming endogenous glial cells into neurons in the central nervous system through ectopically expressing neural transcriptional factors in glial cells. Previous studies have been focusing on glial cells in the grey matter such as the cortex and striatum, but whether glial cells in the white matter can be reprogrammed or not is unknown. To address this fundamental question, we express NeuroD1 in the astrocytes of both grey matter(cortex and striatum) and white matter(corpus callosum) to investigate the conversion efficiency, neuronal subtypes, and electrophysiological features of the converted neurons. We discover that NeuroD1 can efficiently reprogram the astrocytes in the grey matter into functional neurons, but the astrocytes in the white matter are much resistant to neuronal reprogramming. The converted neurons from cortical and striatal astrocytes are composed of both glutamatergic and GABAergic neurons, capable of firing action potentials and having spontaneous synaptic activities. In contrast, the few astrocyte-converted neurons in the white matter are rather immature with rare synaptic events. These results provide novel insights into the differential reprogramming capability between the astrocytes in the grey matter versus the white matter, and highlight the impact of regional astrocytes as well as microenvironment on the outcome of glia-toneuron conversion. Since human brain has large volume of white matter, this study will provide important guidance for future development of in vivo glia-to-neuron conversion technology into potential clinical therapies. Experimental protocols in this study were approved by the Laboratory Animal Ethics Committee of Jinan University(approval No. IACUC-20180321-03) on March 21, 2018.展开更多
This study explored protective effects of Houshiheisan and its compound prescription of wind-dispelling drugs and deficiency-nourishing drugs on cerebral ischemia in terms of astrocyte activation and inflammatory fact...This study explored protective effects of Houshiheisan and its compound prescription of wind-dispelling drugs and deficiency-nourishing drugs on cerebral ischemia in terms of astrocyte activation and inflammatory factor expression.Results suggested that Houshiheisan lessened neuronal degeneration in the corpus striatum on the ischemic side of rats following cerebral ischemia/reperfusion injury,contributed to astrocyte activation and glial fibrillary acidic protein expression in the corpus striatum and decreased the levels of interleukin-2,interleukin-6, interleukin-1βand tumor necrosis factor-α.Factor analysis results demonstrated that deficiency-nourishing drugs were more beneficial in protecting neurons and upregulating glial fibrillary acidic protein expression than wind-dispelling drugs.However,wind-dispelling drugs were more effective in increasing the number of glial fibrillary acidic protein-positive cells and reducing inflammatory factor expression than deficiency-nourishing drugs.These indicate that different ingredients of Houshiheisan suppress cerebral ischemic injury by promoting astrocyte activation and diminishing inflammatory factor expression.展开更多
γ-Aminobutyric acid（GABA）,plays a key role in all stages of life,also is considered the main inhibitory neurotransmitter.GABA activates two kind of membrane receptors known as GABAA and GABAB,the first one is respo...γ-Aminobutyric acid（GABA）,plays a key role in all stages of life,also is considered the main inhibitory neurotransmitter.GABA activates two kind of membrane receptors known as GABAA and GABAB,the first one is responsible to render tonic inhibition by pentameric receptors containing α4-6,β3,δ,or ρ1-3 subunits,they are located at perisynaptic and/or in extrasynaptic regions.The biophysical properties of GABAA tonic inhibition have been related with cellular protection against excitotoxic injury and cell death in presence of excessive excitation.On this basis,GABAA tonic inhibition has been proposed as a potential target for therapeutic intervention of Huntington＇s disease.Huntington＇s disease is a neurodegenerative disorder caused by a genetic mutation of the huntingtin protein.For experimental studies of Huntington＇s disease mouse models have been developed,such as R6/1,R6/2,Hdh Q92,Hdh Q150,as well as YAC128.In all of them,some key experimental reports are focused on neostriatum.The neostriatum is considered as the most important connection between cerebral cortex and basal ganglia structures,its cytology display two pathways called direct and indirect constituted by medium sized spiny neurons expressing dopamine D1 and D2 receptors respectively,they display strong expression of many types of GABAA receptors,including tonic subunits.The studies about of GABAA tonic subunits and Huntington＇s disease into the neostriatum are rising in recent years,suggesting interesting changes in their expression and localization which can be used as a strategy to delay the cellular damage caused by the imbalance between excitation and inhibition,a hallmark of Huntington＇s disease.展开更多
Autapses selectively form in specific cell types in many brain regions.Previous studies have also found putative autapses in principal spiny projection neurons(SPNs)in the striatum.However,it remains unclear whether t...Autapses selectively form in specific cell types in many brain regions.Previous studies have also found putative autapses in principal spiny projection neurons(SPNs)in the striatum.However,it remains unclear whether these neurons indeed form physiologically functional autapses.We applied whole-cell recording in striatal slices and identified autaptic cells by the occurrence of prolonged asynchronous release(AR)of neurotransmitters after bursts of high-frequency action potentials(APs).Surprisingly,we found no autaptic AR in SPNs,even in the presence of Sr^(2+).However,robust autaptic AR was recorded in parvalbumin(PV)-expressing neurons.The autaptic responses were mediated by GABA_(A) receptors and their strength was dependent on AP frequency and number.Further computer simulations suggest that autapses regulate spiking activity in PV cells by providing self-inhibition and thus shape network oscillations.Together,our results indicate that PV neurons,but not SPNs,form functional autapses,which may play important roles in striatal functions.展开更多
In adult mammals, including humans, the subventricular zone （SVZ） of the lateral ventricles and the subgranular zone of the dentate gyms （DG） show ongoing neurogenesis. Cerebral ischemic insults trigger neurogenes...In adult mammals, including humans, the subventricular zone （SVZ） of the lateral ventricles and the subgranular zone of the dentate gyms （DG） show ongoing neurogenesis. Cerebral ischemic insults trigger neurogenesis from neural stem cells or progenitor cells located in the SVZ and DG. Newborn neurons are then functionally recruited into the circuitry of the CA1 region, striatum and DG granule cell layer.展开更多
The striatum, as the primary input nucleus in the basal ganglion,plays an important role in neural circuits crucial for the control of critical motivation, motor planning and procedural learning(Kreitzer and Malenka, ...The striatum, as the primary input nucleus in the basal ganglion,plays an important role in neural circuits crucial for the control of critical motivation, motor planning and procedural learning(Kreitzer and Malenka, 2008). Most cells in the striatum are GABAergic, including a large population (90%-95%) of medium spiny neurons (MSNs) and a small population of interneurons.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.
基金supported by the Natural Science Foundation of Fujian Province, No. 2009J06018
文摘The Chinese herbs Herba Epimedii, Fructus Ligustri Lucidi and Rhizoma Polygonati were injected into Parkinson＇s disease mice established via intraperitoneal injection of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine hydrochloride. The selective monoamine oxidase B inhibitor selegiline was used as a positive control drug. After successive administration for 4 weeks, Herba Epimedii could downregulate the expression of caspase-3 and increase the brain-derived neurotrophic factor level, as well as increase tyrosine hydroxylase activity in the substantia nigra of Parkinson＇s disease mouse models. Rhizoma Polygonaticould downregulate the expression of caspase-3 and FasL, and increase neural growth factor and brain-derived neurotrophic factor levels. Fructus Ligustn Lucidi could downregulate caspase-3 expression. Rhizoma Polygonati and Fructus Ligustn Lucidi did not produce obvious effects on tyrosine hydroxylase activity. Herba Epimedii and Fructus Ligustri Lucidi yielded similar effects on apoptosis-promoting factors to those elicited by selegiline. Herba Epimedii and Rhizoma Polygonati significantly increased the levels of neurotrophic factors compared with selegiline. Herba Epimedii significantly increased tyrosine hydroxylase activity compared with selegiline. It is indicated that the kidney-tonifying Chinese herbal preparation can downregulate the expression of apoptosis-promoting factors, increase neurotrophic factors levels in the substantia nigra and striatum, as well as increase tyrosine hydroxylase activity in the substantia nigra of Parkinson＇s disease mouse models, thereby exerting a stronger or similar neuroprotective effects compared with selegiline.
基金supported by the National Natural Science Foundation of China,No.81072506
文摘The over-expression of α-synuclein is a major factor in the death of dopaminergic neurons in a methamphetamine-induced model of Parkinson’s disease. In the present study, α-synuclein knockdown rats were created by injecting α-synuclein-shRNA lentivirus stereotaxically into the right striatum of experimental rats. At 2 weeks post-injection, the rats were injected intraper-itoneally with methamphetamine to establish the model of Parkinson’s disease. Expression of α-synuclein mRNA and protein in the right striatum of the injected rats was significantly down-regulated. Food intake and body weight were greater in α-synuclein knockdown rats, and water intake and stereotyped behavior score were lower than in model rats. Striatal dopamine and tyrosine hydroxylase levels were significantly elevated in α-synuclein knockdown rats. Moreover, superoxide dismutase activity was greater in α-synuclein knockdown rat striatum, but the levels of reactive oxygen species, malondialdehyde, nitric oxide synthase and nitrogen monoxide were lower compared with model rats. We also found that α-synuclein knockdown inhibited metham-phetamine-induced neuronal apoptosis. These results suggest that α-synuclein has the capacity to reverse methamphetamine-induced apoptosis of dopaminergic neurons in the rat striatum by inhibiting oxidative stress and improving dopaminergic system function.
基金supported in part by the National Natural Science Foundation of China(Grant No.31701291 to WL,U1801681 to GC)the China Postdoctoral Science Foundation(Grant No.2016M602600 to WL)+1 种基金the Guangdong Grant ‘Key Technologies for Treatment of Brain Disorders’(Grant No.2018B030332001 to GC)the Internal Funding of Jinan University,China(Grant No.21616110 to GC)
文摘A new technology called in vivo glia-to-neuron conversion has emerged in recent years as a promising next generation therapy for neural regeneration and repair. This is achieved through reprogramming endogenous glial cells into neurons in the central nervous system through ectopically expressing neural transcriptional factors in glial cells. Previous studies have been focusing on glial cells in the grey matter such as the cortex and striatum, but whether glial cells in the white matter can be reprogrammed or not is unknown. To address this fundamental question, we express NeuroD1 in the astrocytes of both grey matter(cortex and striatum) and white matter(corpus callosum) to investigate the conversion efficiency, neuronal subtypes, and electrophysiological features of the converted neurons. We discover that NeuroD1 can efficiently reprogram the astrocytes in the grey matter into functional neurons, but the astrocytes in the white matter are much resistant to neuronal reprogramming. The converted neurons from cortical and striatal astrocytes are composed of both glutamatergic and GABAergic neurons, capable of firing action potentials and having spontaneous synaptic activities. In contrast, the few astrocyte-converted neurons in the white matter are rather immature with rare synaptic events. These results provide novel insights into the differential reprogramming capability between the astrocytes in the grey matter versus the white matter, and highlight the impact of regional astrocytes as well as microenvironment on the outcome of glia-toneuron conversion. Since human brain has large volume of white matter, this study will provide important guidance for future development of in vivo glia-to-neuron conversion technology into potential clinical therapies. Experimental protocols in this study were approved by the Laboratory Animal Ethics Committee of Jinan University(approval No. IACUC-20180321-03) on March 21, 2018.
基金funded by the National Natural Science Foundation of China,No.30973782the Natural Science Foundation of Beijing,No.7102014+1 种基金the Science and Technology Program of Chinese Medicine of Beijing City,No.JJ2008-042the Chinese Medicine Nursing Special Foundationof Beijing Education Commission,No.10ZYH04
文摘This study explored protective effects of Houshiheisan and its compound prescription of wind-dispelling drugs and deficiency-nourishing drugs on cerebral ischemia in terms of astrocyte activation and inflammatory factor expression.Results suggested that Houshiheisan lessened neuronal degeneration in the corpus striatum on the ischemic side of rats following cerebral ischemia/reperfusion injury,contributed to astrocyte activation and glial fibrillary acidic protein expression in the corpus striatum and decreased the levels of interleukin-2,interleukin-6, interleukin-1βand tumor necrosis factor-α.Factor analysis results demonstrated that deficiency-nourishing drugs were more beneficial in protecting neurons and upregulating glial fibrillary acidic protein expression than wind-dispelling drugs.However,wind-dispelling drugs were more effective in increasing the number of glial fibrillary acidic protein-positive cells and reducing inflammatory factor expression than deficiency-nourishing drugs.These indicate that different ingredients of Houshiheisan suppress cerebral ischemic injury by promoting astrocyte activation and diminishing inflammatory factor expression.
基金the programs for the postdoctoral fellowships-Chilean CONICYT-FONDECYT#3140218,Mexican CONACYT#164978 and DID-UACh S-2015-81Sistema Nacional de Investigadores#58512 to Abraham Rosas-Arellano+2 种基金supported by USACH PhD fellowshipsupported with a PhD fellowship from CONACYT(#299627)FONDECYT grants 1151206 and 1110571 to Maite A.Castro
文摘γ-Aminobutyric acid（GABA）,plays a key role in all stages of life,also is considered the main inhibitory neurotransmitter.GABA activates two kind of membrane receptors known as GABAA and GABAB,the first one is responsible to render tonic inhibition by pentameric receptors containing α4-6,β3,δ,or ρ1-3 subunits,they are located at perisynaptic and/or in extrasynaptic regions.The biophysical properties of GABAA tonic inhibition have been related with cellular protection against excitotoxic injury and cell death in presence of excessive excitation.On this basis,GABAA tonic inhibition has been proposed as a potential target for therapeutic intervention of Huntington＇s disease.Huntington＇s disease is a neurodegenerative disorder caused by a genetic mutation of the huntingtin protein.For experimental studies of Huntington＇s disease mouse models have been developed,such as R6/1,R6/2,Hdh Q92,Hdh Q150,as well as YAC128.In all of them,some key experimental reports are focused on neostriatum.The neostriatum is considered as the most important connection between cerebral cortex and basal ganglia structures,its cytology display two pathways called direct and indirect constituted by medium sized spiny neurons expressing dopamine D1 and D2 receptors respectively,they display strong expression of many types of GABAA receptors,including tonic subunits.The studies about of GABAA tonic subunits and Huntington＇s disease into the neostriatum are rising in recent years,suggesting interesting changes in their expression and localization which can be used as a strategy to delay the cellular damage caused by the imbalance between excitation and inhibition,a hallmark of Huntington＇s disease.
基金supported by the National Natural Science Foundation of China(32130044,31630029,32171094,and 32100930)the National Key Research and Development Program of China(2021ZD0202500).
文摘Autapses selectively form in specific cell types in many brain regions.Previous studies have also found putative autapses in principal spiny projection neurons(SPNs)in the striatum.However,it remains unclear whether these neurons indeed form physiologically functional autapses.We applied whole-cell recording in striatal slices and identified autaptic cells by the occurrence of prolonged asynchronous release(AR)of neurotransmitters after bursts of high-frequency action potentials(APs).Surprisingly,we found no autaptic AR in SPNs,even in the presence of Sr^(2+).However,robust autaptic AR was recorded in parvalbumin(PV)-expressing neurons.The autaptic responses were mediated by GABA_(A) receptors and their strength was dependent on AP frequency and number.Further computer simulations suggest that autapses regulate spiking activity in PV cells by providing self-inhibition and thus shape network oscillations.Together,our results indicate that PV neurons,but not SPNs,form functional autapses,which may play important roles in striatal functions.
文摘In adult mammals, including humans, the subventricular zone （SVZ） of the lateral ventricles and the subgranular zone of the dentate gyms （DG） show ongoing neurogenesis. Cerebral ischemic insults trigger neurogenesis from neural stem cells or progenitor cells located in the SVZ and DG. Newborn neurons are then functionally recruited into the circuitry of the CA1 region, striatum and DG granule cell layer.
基金supported by grants from the National Key Research and Development Program of China (2016YFA0100702 and 2016YFC0902502)the National Natural Science Foundation of China (31670789 and 31671316)the CAMS Innovation Fund for Medical Sciences (CIFMS, 2016I2M-2-001, 2016-I2M-1-001, 2016-I2M-1-004 and 2017-I2M-1004)
文摘The striatum, as the primary input nucleus in the basal ganglion,plays an important role in neural circuits crucial for the control of critical motivation, motor planning and procedural learning(Kreitzer and Malenka, 2008). Most cells in the striatum are GABAergic, including a large population (90%-95%) of medium spiny neurons (MSNs) and a small population of interneurons.
