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Astrocytes. a New Target that Unlocks Brain Disorder Secrets and Drug Discoveries

中国药理通讯 2013年第3期30卷 8-9页

作者： albert cheung hoi yu Neuroscience Research Institute Key Laboratory for Neuroscience (Ministry of Education) Key Laboratory for Neuroscience (National Health and Family Planning Commission) Department of Neurobiology School of Basic Medical Sciences Health Science Center Peking University Beijing 100191 China

Astrocytes are the most abundant cells in the brain. Increasing evidence indicates that astrocytes are not ' brain glue~ , but are indeed critical for various functions in the central nervous systems (CNS), including ... 详细信息

关键词：论文摘要 "第十三届全国生化与分子药理学学术会议" 报告 PISD

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Quantitative measurement of glutamate uptake by isocratic reversed high performance liquid chromatography

Quantitative measurement of glutamate uptake by isocratic re...

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中国神经科学学会第四次会员代表大会暨第七届全国学术会议

作者： albert cheung hoi yu Neuroscience Research Institute Department of Neurobiology School of Basic Medical Sciences Key Laboratory for Neuroscience Ministry of Education Key Laboratory for Neuroscience Ministry of Public HealthPeking University

＜正＞In the central nervous system,neuronal-released glutamate is mainly uptaken by ***,the fate of uptaken glutamate is still *** used isocratic reversed high performance liquid chromatography（RPHPLC） to detect the change in contents of glutamate（Glu）,glutamine（Gln） and taurine（Tau） in both intact astrocytes and their isolated nuclei after Glu *** resolution of these amino acids was performed in an Allsphere ODS-2 C18[250±4.6) cm,5μm]*** composition of mobile phase was 100 mmol/L acetic buffer,pH 5-6.5 containing 20%methanol and 10%acetonitrile at a flow rate of 1.0 ml/*** used homoserine as an internal standard control. The fluorimetric detector was set at 340 nm Ex and 445 nm *** time for Glu,Gln and Tau were 3.02 min,4.7 min and 8.8 min,*** curves were linear（r = 0.9998） in the range of 0.0005-100 mmol/L for all three amino *** results showed that Glu,but not Gln and Tau,increased significantly in both intact astrocytes and their isolated nuclei after 1 mM Glu *** indicated that Glu released from neurons was not only taken by astrocyte into cytoplasm but also into its nucleus.

关键词： RP-HPLC pre-columndrivation glutamate glutamine taurine astrocyte fluorimetric detection

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Aquaporin 1 specifically accumulate around astrocyte nuclear to mediates Glu-induced nuclear swelling

Aquaporin 1 specifically accumulate around astrocyte nuclear...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： albert cheung hoi yu Neuroscience Research Institute Peking University Key Laboratory for Neuroscience（Peking University） Ministry of Education Key Laboratory for Neuroscience（Peking University） Ministry of Health Department of Neurobiology Peking University Health Science Center Infectious Disease Center Peking University laboratory of Translational Medicine Institute of Systems BiomedicinePeking University

＜正＞Objective Aquaporins（AQPs）belong to the water-specific channel protein family and have been reported to involve in cell and its organelle *** has been reported to involve in multiple functions in *** study is to explore the role of AQPl in the specific nuclear response of astrocytes to *** Expression and localization of AQPl in atrocytes were detected through RT-PCR,Western Blot,Immunostaining and AQPl-EGFP *** of AQPl and AQP1’s inhibitor treatment were used to study the role of AQPl in Gluinduced nuclear swelling in *** We found that aquaporin-1（AQPl）was uniquely expressed and located on the nuclear membrane of astrocytes to facilitate Glu-induced nuclear ***-expression of AQP1 in astrocytes could accelerate and enhance nuclear swelling and if inhibited would prevent *** AQPl localization and Glu-induced nuclear swelling are both specific to astrocytes,and could not be found in any other cells *** unique features were also observed in human *** Our study revealed that AQPl mediated a specific nuclear response of astrocytes to Glu that might serve to facilitate communication of excitatory information between neurons and astrocytes through an as yet unknown mechanism.

关键词： aquaporin-1 glutamate astrocyte nuclear swelling

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Astrocytes expresses HCN channel which responds to K+ and glutamate

Astrocytes expresses HCN channel which responds to K+ and gl...

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中国神经科学学会第四次会员代表大会暨第七届全国学术会议

作者： albert cheung-hoi yu Neuroscience Research Institute Department of Neurobiology School of Basic Medical Sciences Key Laboratory for Neuroscience Ministry of Education Key Laboratory for Neuroscience Ministry of Public HealthPeking University

We recently found astrocytes in primary culture expressed hyperpolarization-activated cyclic nucleotide-gated (HCN) genes(HCN2 and HCN4) and the hyperpolarization-activated cation current(I).The expressions of HCN genes and I currents were different between astrocytes and *** we demonstrated that both increase and decrease in [K] would down-regulate the gene expression *** of osmolality and sodium content in the medium did not affect HCN2 gene *** of[K] can significantly up-regulate I *** treatment alone did not affect the HCN2 gene expression and I current in ***-treatments of glutamate and K could modulate the effect of K to the HCN2 gene expression and the glutamate effect was mediated by AMPA and NMDA *** findings indicate that the astrocytic HCN channel could respond to K and glutamate,2 major factors in neuronal excitation. Therefore,HCN might play an important role in controlling the resting membrane potential of astrocytes,thus contributing to the maintenance of electrophysiological homeostasis of the brain.

关键词： HCN astrocyte K+ glutamate

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High-potassium induced up-regulation of aquaporin-9 in primary culture of astrocyte

High-potassium induced up-regulation of aquaporin-9 in prima...

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中国神经科学学会第六届学术会议暨学会成立十周年庆祝大会

作者： albert cheung-hoi yu Neuroscience Research Institute Peking University Key Laboratory of Neuroscience （Peking University） Ministry of Education Department of Neurobiology Peking University Health Science Center

＜正＞Aquaporins （AQPs） are channel proteins that mediate water transportation through plasma membrane. This group of ubiquitous proteins is demonstrated to have their essential functions in maintaining water homeostasis, regulating cell volume in nearly all types of tissues. In the central nervous system, AQP4 and AQP9 are the most abundant expressed subtypes among AQP family. Research indicates that AQP4 and AQP9 may conduct essential roles in constructing blood-brain-barrier and

关键词： aquaporin-9 potassium astrocyte

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Ischemia caused JNK activation to induce 14-3-3g up-regulation in astrocyte

Ischemia caused JNK activation to induce 14-3-3g up-regulati...

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中国神经科学学会第四次会员代表大会暨第七届全国学术会议

＜正＞Our laboratory has previously reported that 14-3-3g could be specifically induced by ischemia in astrocyte and has a neuroprotective *** attempted to uncover the mechanism involved in this *** primary culture of astrocyte,we investigated the involvement of the three major components of ischemia（glucose deprivation,hypoxia and toxin accumulation） in the 14-3-3g *** deprivation alone did not cause a detectable change in the 14- 3-3g mRNA and protein levels,while hypoxia and toxin accumulation could slightly elevate 14-3-3g expression,but the degree of their effects were less and later than *** further investigated whether the ischemia induced activation of PI3K/Akt and MAPK signaling pathways we have previously reported in our laboratory has any role in 14-3-3g up-regulation. Using various pathway inhibitors（LY294002 for PI3K,U0126 for ERK,SB203580 for p38 and SP600125 for JNK）, we found that only JNK inhibitor（SP600125） could prevent the 14-3-3g up-regulation in astrocyte under *** results demonstrated that 14-3-3g up-regulation in astrocyte under ischemia was mediated by JNK activation.

关键词： 14-3-3g astrocyte ischemia JNK

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GFAP upregulation in mechanical injury depends on calcium-induced activation of JNK/c-Jun/AP-1 signal pathway

GFAP upregulation in mechanical injury depends on calcium-in...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： albert cheung hoi yu Neuroscience Research Institute Peking University Key Laboratory of Neuroscience(Peking University) Ministry of Education Key Laboratory of Neuroscience(Peking University) Ministry of Health Department of Neurobiology Peking University Health Science Center Infectious Disease Center Peking University Laboratory of Trans lational Medicine Institute of Systems BiomedicinePeking University

Objective Overexpression of glial fibrillary acidic protein(GFAP) is a hallmark of astrogliosis induced by injury and disease in the central nervous *** mechanism of GFAP upregulation in reactive astrocytes still remains to be *** study is to explore how GFAP is upregulated after astrocyte reactivation. Methods In vitro scratch injury model was used in this *** of GFAP was detected by Real-Time PCR and Western *** of related signaling pathway were used to study the mechanism of GFAP upregulation. Results After scratch injury,we found c-Jun N-terminal kinase(JNK) was *** the p-JNK phosphorylated the putative transcriptional factor c-Jun which facilitated AP-1 binding site on GFAP gene promoter region to upregulate GFAP *** of JNK by SP600125 significantly inhibited the levels of GFAP mRNA and *** also demonstrated that calcium was involved in the activation of *** Ca chelator BAPTA-AM could inhibit the JNK activation and GFAP *** Taken together,our findings indicate that the Ca/JNK/c-Jun/AP-1 pathway regulates GFAP during astrocyte reactivation.

关键词： astrogliosis GFAP JNK calcium

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Glutamate induces astrocytic nuclear swelling through aquaporin-1 in nuclear membrane

Glutamate induces astrocytic nuclear swelling through aquapo...

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中国神经科学学会第四次会员代表大会暨第七届全国学术会议

＜正＞In the central nervous system,glutamate is mainly released as excitatory neurotransmitter from *** injury and many pathological conditions,huge amount of glutamate is *** uptake is the major mechanism to remove glutamate from the *** previous findings showed that the uptaken glutamate could be synthesized to glutamine and utilized by the TCA ***,there is a lack of realization on how astrocyte recognizes neuronalreleased glutamate under *** we reported that in addition to the well-known cytosolic swelling,glutamate could trigger astrocytic nuclear *** nuclear swelling was independent from the cytosolic *** appeared to be a direct effect of glutamate and was concentration *** induced nuclear swelling may be astrocytic specific as we did not observe a similar effect in *** requires the glutamate transporters on the plasma membrane and the aquaporin-1 on the astrocytic nuclear *** findings discovered that astrocyte recognizes neuronal excitation through nuclear swelling induced by the uptaken *** might be an important form of interaction between neurons and astrocytes during neuronal excitation especially under various pathological conditions.

关键词： glutamate astrocyte nuclear swelling glutamate uptake

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Characterization of DREAM isoforms in astrocytes and neurons

Characterization of DREAM isoforms in astrocytes and neurons

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

Objective Downstream regulatory element antagonist modulator(DREAM),as a multifunctional Ca binding protein,binds with specific DNA sequence to regulate gene expression and with Kv4.2 or presenilins outside the nucleus to regulate membrane excitability and calcium homeostasis,*** main isoforms of DREAM (A and B) were produced by alternative *** study is to observe the characterization of DREAM isoforms expression and distribution in astrocytes and *** Expression of DREAM A and B mRNA in primary culture of neural cells(astrocytes,GABAergic neuron and glutamatergic neuron) was measured by real time PCR. Subcloned DREAM isoform A or B in pIRES2-EGFP vectors and the plasmids were transfected into astrocytes to observe their subcellular *** We found that both DREAM A and B are present in all neural cells *** A was in a higher level in glutamatergic neuron whereas isoform B was in a higher level in astrocyte. Isoform A showed a diffusible distribution and isoform B appeared mainly localized in the *** mRNA levels of these two isoforms were measured in these neural cells at different *** found that isoform A showed the highest level in older but isoform B showed the highest level in younger *** confirmed these developmental changes in tissue of cerebral cortex and *** These data suggest that DREAM A is the major isoform in mature cells and can function as a transcriptional *** B is mainly localized in ER and may play a role in the early stage of development.

关键词： astrocyte neuron DREAM isoforms

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A new specialization in astrocytes——Glutamate induces nuclear size change through aquaporin 1

A new specialization in astrocytes——Glutamate induces nucl...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

＜正＞Objective Astrocytes play an important role in the homeostasis of Glu during excitatory information transmission in the *** could utilize the Glu transporters to remove Glu from the synaptic *** astrocytes,the internalized Glu is utilized for glutamine synthesis through glutamine synthetase,or for energy production through the TCA ***,we estimated that the total amount of Glu uptaken into astrocytes is much higher than the combined amount of Glu dedicated to glutamine synthesis and TCA *** would result in an accumulation of Glu in the *** the vital roles of Glu as the most important signaling molecule in the CNS,it is unlikely that the cytoplasmic Glu remains dormant and merely waits to be *** reported that cytoplasmic Glu could enter nucleus and cause nuclear *** Morphological changes on various cell types and their isolated nuclei were observed by phase contrast microscopy,fluorescence microscopy,confocal laser microscopy and atomic force *** and agonists of different Glu receptors,inhibitors of Glu transporters were used to study the mechanism of Glu-induced nuclear swelling in *** of water channels including AQP1 in astrocyte was determined by RT-PCR,Western Blot,immunostaining and AQP1-EGFP *** Glu was found to specifically induce reversible nuclear swelling in astrocytes,but not in any other cell types and their isolated *** was a direct and specific effect limited to Glu but not other related *** is metabolically related to Glu and could induce an indirect nuclear swelling in intact astrocytes but a direct shrinkage in isolated *** unique features are required for this Glu-induced nuclear swelling:the Glu transporters on plasma membrane and the specific expression of AQP1 on the nuclear membrane of astrocytes. In addition,similar Glu-induced nuclear swelling and specific AQP1 nuclear localization

关键词： glutamate astrocyte aquaporin-1 nuclear swelling

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