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Expression of N-methyl-D-aspartic acid 2A-B and 2B receptors in anterior thalamic nucleus and subiculum complex of rats

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Neural Regeneration Research 2008年第8期3卷 822-825页

作者： Yuanshan Fu Xiaokai Ma Xiaoling Yue Bin Wang Department of Anatomy Dalian Medical University Dalian 116044 Liaoning Province China

BACKGROUND：Glutamate acid ionotropic receptor N-methyl-D-aspartic acid （nmda） takes part in long-term potentiation, thereby influencing the process of learning and memory. OBJECTIVE： To verify expression of nmda 2A/B and 2B receptors in the anterior thalamic nucleus and subiculum complex of rats. DESIGN, TIME AND SETTING： A single-sample observation was performed at Department of Anatomy in Dalian Medical University （Dalian, Liaoning, China） from April to September in 2007. MATERIALS： Ten adult Wistar rats were used for this study, as well as rabbit anti-nmda 2A/B and 2B antibodies. METHODS： The rats were anesthetized and perfused, followed by brain resection and coronal sectioning of the brain tissue. A 1：3 series was selected for immunohistochemistry, using antibodies specific to nmda 2A/B and 2B receptors. Photos were taken using the Nikon image analysis system. MAIN OUTCOME MEASURES： Expression and distribution of immunohistochemistry staining of nmda 2A/B and 2B receptor subunits. RESULTS： There were a large number of nmda 2A/B and 2B receptor-positive neurons distributed throughout the anterior dorsal thalamic nucleus. In the anterior ventral thalamic nucleus, distribution of positive neurons was rare, staining intensity was lighter, and cell bodies were smaller compared with the anterior dorsal thalamic nucleus. In the subiculum complex, staining intensity of nmda 2A/B and 2B-positive neurons was weakest in the molecular layer and stronger in the pyramidal layer, in particular the region with large cell bodies adjacent to the molecular layer. In the multiform layer, more positive neurons of various sizes were detected. CONCLUSION： nmda 2A/B and 2B receptor subunits were richly distributed in the anterior thalamic nucleus, with a small difference existing between the anterior dorsal nucleus and anterior ventral nucleus. These neurons were also differentially distributed within the three layers of the subiculum complex.

关键词： anterior thalamic nucleus learning memory nmda receptor rats subiculum complex

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Induction of Increased Intracellular Calcium in Astrocytes by Glutamate through Activating nmda and AMPA receptors

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2003年第3期23卷 254-257页

作者：张蕲胡波孙圣刚童萼塘 Department of Pathophysiology Department of Neurosurgery

To study the effect of glutamate on the intracellular calcium signal of pure cultured rat astrocytes and the role of nmda and AMPA receptors in the procedure, the change of calcium signal was investigated by monitoring the fluctuation of intracellular Ca 2+ concentration ([Ca 2+ ] i) on the basis of Fura-2 single cell fluorescent ratio (F345/F380). The changes in the effect of glutamate on the intracellular calcium signal were observed after blockage of nmda and(or) AMPA receptors. It was found that L-glutamate could induce an increased [Ca 2+ ] i in most of the cells in concentration- and time-dependent manner. D-(-)-2-amino-5-phosphonopentanoic acid (D-AP-5, a selective antagonist of the nmda receptor) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, a selective antagonist of the AMPA receptor) could abolish the effects of nmda and AMPA respectively. The treatment of D-AP-5 and CNQX simultaneously or respectively could attenuate the effect of L-glutamate at varying degrees. All these indicated that glutamate could modulate intracellular Ca 2+ of pure cultured rat astrocytes through different pathways. The activation of nmda and AMPA receptors took part in the complex mechanisms.

关键词： glutamate astrocytes nmda receptor AMPA receptor [Ca 2+] i

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Neuroprotective strategies for nmdaR-mediated excitotoxicity in Huntington＇s Disease

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Frontiers in Biology 2016年第6期11卷 439-458页

作者： Kimberly D. Girling Yu Tian Wang Djavad Mowafahian Centre for Brain Health & Department of Medicine University of British Columbia V6T 1Z3 Canada

BACKGROUND： Huntington＇s Disease （HD） is an autosomal dominant neurodegenerative disease causing severe neurodegeneration of the striatum as well as marked cognitive and motor disabilities. Excitotoxicity, caused by overstimulation of nmda receptors （nmdaRs） has been shown to have a key role in the neuropathogenesis of liD, suggesting that targeting nmdaR-dependent signaling may be an effective clinical approach for HD. However, broad nmdaR antagonists are generally poor therapeutics in clinical practice. It has been suggested that GluN2A-containing, synaptically located nmdaRs activate cell survival signaling pathways, while GluN2B-containing, primarily extrasynaptic nmdaRs trigger cell death signaling. A better approach to development of effective therapeutics for HD may be to target, specifically, the cell-death specific pathways associated with extrasynaptic GluN2B nmdaR activation, while maintaining or potentiating the cell- survival activity of GluN2A-nmdaRs. OBJECTIVE： This review outlines the role of nmdaR-mediated excitotoxicity in HD and overviews current efforts to develop better therapeutics for HD where nmdaR excitotoxicity is the target. METHODS： A systematic review process was conducted using the PubMed search engine focusing on research conducted in the past 5-10 years. 235 articles were consulted for the review, with key search terms including ＂Huntington＇s Disease,＂＂excitotoxicity,＂＂nmdaR＂ and ＂therapeutics.＂ RESULTS： A wide range of nmdaR excitotoxicity-based targets for HD were identified and reviewed, including targeting nmdaRs directly by blocking GluN2B, extrasynaptic nmdaRs and/or potentiating GluN2A, synaptic nmdaRs, targeting glutamate release or uptake, or targeting specific downstream cell-death signaling of nmdaRs. CONCLUSION： The current review identifies nmdaR-mediated excitotoxicity as a key player in HD pathogenesis and points to various excitotoxicity-focused targets as potential future preventative therapeutics for HD.

关键词： Huntington＇s Disease nmda receptor excitotoxicity cell death therapeutics

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Inhibitory effect of ketamine on lighting amygdala of rats

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Neural Regeneration Research 2006年第2期1卷 130-132页

作者： Jiguo Zhang Bin Yang Jing Zhang Feng Zhang Wang Yue Depallment of Pharmacology Pharmacological College Tais-hart Medical College Taian271016 Shandong ProvinceChina Basic Faculty Depallment of Management TaishanMedical College Taian271016 Shandong ProvinceChina Faculty of Pharmacology Medical College of QingdaoUniversity Qingdao 266021Shandong Province China

BACKGROUND： Ketamine is a noncompetitive antagonist of N-methyI-D-aspartic acid receptor. Some researchers suggest that N-methyI-D-aspartic acid （nmda） receptor is closely related to epileptic attack. OBJECTIVE： To observe inhibitory effect of ketamine on lighting amygdala of rats and analyze pathway of anti-lighting. DESIGN： Randomized controlled animal study SETTING： Department of Pharmacology and Department of Management, Pharmacological College of Taishan Medical College; Department of Pharmacology, Medical College of Qingdao University. MATERIALS： Sixty adult female Wistar rats, of clean grade, weighing 180-200 g, were provided by Animal Center of Qingdao Institute of Drug Control. Ketamine hydrochloride was provided by the First Pharmacological Factory of the First Biochemical Pharmacology Company of Shanghai, and nicardipine, an antagonist of calcium ions, was provided by Sigma Company. METHODS： The experiment was completed in the Department of Pharmacology, Medical College of Qingdao University from March to November 2004. ① Model establishing： After anesthesia, left and right amygdalas were inserted with double electrodes. The top was separated about 0.25 mm, and the other end was welded with a micro-plug, respectively. Electrode and micro-plug were fixed with dental base acrylic resin powder at the surface of cranium. Two weeks after recovery, right amygdala was stimulated with constant current once a day. According to Racine technique, attacking intensity was divided into 5 grades： grade I： closing eyes, a little tingling of beards and twitching face; grade Ⅱ ： nodding, chewing accompanying with twitching face; grade Ⅲ ： raising one of a forelimb and clonus; grade Ⅳ： standing accompanying with bilateral forelimbs; grade Ⅴ： standing accompanying with falling down. Rats with grades Ⅳ and Ⅴ were used to establish secondarily generalized epilepsy. If 3 successive attacks of grade V were observed, the lighting was to be successful. ② Effect of ketamine on lighting amygdala of rats： A total of 24 rat

关键词： constant current inhibitory effect nmda receptor acrylic resin calcium ions animal study adult female Wistar rats

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Salicylate enhances expression and function of nmda receptors in cochlear spiral ganglion neurons

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Journal of Otology 2012年第1期7卷 9-14页

作者： GAO Meng FANG Xiu-you FENG Shuang WANG Ren-jun DENG Li-li SU Ji-ping Department of Otolaryngology Head and Neck Surgery Guangxi Medical UniversityNanningGuangxi 530021 Department of Otolaryngology Head and Neck Surgery Binzhou People HospitalBinzhouShandong 256600 Department of 4^(th)Surgery Ganyu People HospitalLianyugangJiangsu 222100

Objective To study the effect of salicylate on the expression and function of nmda receptors in spiral ganglion neurons （SGNs）. Methods The mRNA of NR1 subunit of nmda receptor in modiolus tissues were detected by Real time fluorescence quantitative PCR （FQ-PCR）. nmda receptor whole-cell currents were recorded using patch clamp in acute isolated SGNs. Results Compared with the control group, salicylate significantly increased the mRNA level of NR1 subunit in SGNs. nmda of concentrations ranging from 0.1 mM to 10 mM evoked no current in SGNs. nmda （0. 1mM and 0.5 mM） applied with salicylate （5 mM）, however, induced inward currents （212.6±15.2pA, n=5; 607.9±44.3pA, n=5） in a dose-dependent manner, which could be inhibited by APV. Salicylate alone did not produce any current in SGNs. Conclusion Salicylate increases the expression of nmda receptors and facilitates the currents mediated by nmda receptors in SGNs.

关键词： Salicylate nmda receptor spiral ganglion neurons NR1 subunit

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Recent development in nmda receptors

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Chinese Medical Journal 2000年第10期113卷 84-92页

作者：刘云张均田 National Institute of Mental Health Department of Pharmacology Institute of Materia MedicaChinese Academe of Medical Sciences

Purpose To identify the structure and the function of nmda receptors, to understand the modulatory mechanism of some endogenous and exogenous compounds on nmda receptors, and to provide theoretical basis for developing new drugs that modulate nmda receptors Data sources A total of 24 originally identified articles were selected Study selection A total of 24 articles were selected from several hundred original articles or reviews. The content of selected articles are in accordance with our purpose and the authors are authorized scientists in the study on nmda receptors.Data extraction After careful review of the selected papers, the meaningful results and conclusions were extracted using scientific criteria and our experience in the research of nmda receptors.Results nmda receptor contains at least five subunits. They were designated as the NR1 (ζ1), NR2A (ε1), NR2B (ε2), NR2C (ε3), and NR2D (ε4). A unique feature of nmda receptor is the requirement for both glutamate and the co agnist glycine for the efficient gating. nmda receptor is modulated by a number of endogenous and exogenous compounds. Mg 2+ not only blocks the nmda channel in a voltage dependent manner but also potentiates nmda induced responses at positive membrane potentials. Na +, K + and Ca 2+ not only pass through the nmda receptor channel but also modulate the activity of nmda receptors. Zn 2+ blocks the nmda current in a noncompetitive and a voltage independent manner. It has been demonstrated that polyamines do not directly activate nmda receptors, but instead act to potentiate or inhibit glutamate mediated responses. The activity of nmda receptors is also strikingly sensitive to the changes in H + concentration, and partially inhibited by the ambient concentration of H + under physiological conditions.Conclusions nmda receptors are glutamate regulated by ion channels that are permeable to Ca 2+ , Na +, K + and are sensitive to voltage dependent Mg 2+ block This channel complex contributes to excit

关键词： nmda receptor ion-channel cation amino acid

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β-Adrenergic activation enhances nmda-induced current in pyramidal cells of the basolateral nucleus of amygdala

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Science Bulletin 2005年第17期 152-154页

作者： LIU Xinqiu, CAO Xiaohua & LI Bao-ming Laboratory of Higher Brain Functions, Institute of Neurobiology, Insti-tute of Brain Science, Fudan University, Shanghai 200433, China

nmda receptor (nmda-R) in the amygdala complex is critical for both long-term potentiation (LTP) and formation of conditioned fear memory. It is reported that activation of β-adrenoceptors (β-AR) in the amygdala facili-tates LTP and enhances memory consolidation. The present study examined the regulatory effect of β-AR activation on nmda-R mediated current in pyramidal cells of the baso-lateral nucleus of amygdala (BLA), using whole-cell re-cording technique. Bath application of the β-AR agonist iso-proterenol enhanced nmda-induced current, and this fa-cilitatory effect was blocked by co-administered propranolol, a β-AR antagonist. The facilitatory effect of isoproterenol on nmda-induced current could not be induced when the pro-tein kinase A (PKA) inhibitor Rp-cAMPs was added in elec-trode internal solution.The present results suggest that β-AR activation in the BLA could modulate nmda-R activity di-rectly and positively, probably via PKA.

关键词： β-adrenoceptor nmda receptor amygdala basolateral nucleus of amygdala rat.

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Lithium decreased NR2B tyrosine phosphorylation and interactions of NR2B and PSD-95 with Src in rat hippocampus following cerebral ischemia

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Journal of Nanjing Medical University 2005年第5期19卷 256-260页

作者： Juan Ma Xuebo Yan Guangyi Zhang Department of pathology Nanjing Medical University Nanjing 210029 China Research Center for Biochemistry and Molecular Biology Xuzhou Medical College Xuzhou 221002 China

Objective： To study the effects of chronic lithium on N-methyl-d-aspartate receptor subunit 2B （NR2B） tyrosine phosphorylation and the interactions of NR2B and PSD-95 with Src induced by cerebral ischemia/reperfusion （I/R）. Methods： Transient （15min） cerebral ischemia was induced by four-vessel occlusion procedure in SD rats. Immunoprecipitation （IP） and immunoblotting （IB） were performed to investigate the phosphorylation and interactions of proteins. The effects of lithium on tyrosine phosphorylation of NR2B and its interactions with PSD-95 and Src were examined. Results： Transient cerebral ischemia 15 rain followed by reperfusion 6h （I/R 6h） caused a significant increase in tyrosine phosphorylation of NR2B. Administration of LiCI for 7days before ischemia caused a profound decrease in tyrosine phosphorylation of NR2B. Similiarly. the interactions of NR2B and PSD-95 with Src were also enhanced by I/R 6h. moreover, these interactions were also inhibited by chronic lithium. Conclusion： Pretreatment with lithium decrease tyrosine phosphorylation of NR2B and interactions of NR2B and PSD-95 with Src during cerebral I/R.

关键词： LiCI nmda receptor ischemia/reperfusion tyrosine phosphorylation

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Effects of galanin on glutamate-mediated secretion from locus coeruleus neurons in rat pontine slices

Effects of galanin on glutamate-mediated secretion from locu...

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2008北京国际生理科学学术大会

作者： ZHANG ClaireXi Institute of Molecular Medicine Peking University

The locus coeruleus(LC)contains the major group of noradrenergic cell bodies projecting to virtually all parts of the central nervous system.Our previous observations showed that several stimuli,including high-K, glutamate and voltage pulses,induce somatic noradrenalin(NA)secretion from LC neurons.Galanin is a 29-amino acid neuropeptide and is important for learning and memory and mood control.Most LC neurons are galaninpositive. Galanin induces a decrease in firing rate and hyperpolarization of LC neurons.It is not clear,however, whether galanin has effects on nmda receptor-mediated somatodendritic secretion from LC neurons.Here,by using combined measurements of amperometry and patch-clamp,we studied whether galanin modulate NA secretion in LC.We found that application of galanin did not induce an amperometric response,while galanin reduced glutamate-mediated somatodendritic secretion from LC neurons.

关键词： locus coeruleus（LC） nmda receptor somatic noradrenalin（NA） amperometric response

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Research on the relationship between NR3A subunit of nmdaR and brain hypoxia and ischemia

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中国药理学与毒理学杂志 2012年第3期 433-433页

作者：王卉 Institute of Pharmacology and Toxicology Academy of Military Medical Sciences

N-methyl-D-aspartate receptors(nmdaR) are glutamate gated ion channels mediating the vast majority of excitatory neurotransmission in central neural system.NR3 is recently discovered subunits of nmdaR,and has been shown to reduce the calcium permeability when is involved in nmdaR.Since Ca2+ influx through nmdaR-operated channels is considered as predominant reason to induce excitotoxicity,NR3 subunits may provide a protective role by depressing calcium overloading.In the present study,it was found that the expression of NR3A protein on rat hippocampus began to increase on the postnatal days,which reached the highest expression level on post natal day 10,and decreased gradually,which remained low into the adulthood.NR3A protein expression was increased in the models of two vessel occlusion rat brain ischemia and cell oxygen and glucose deprivation.The protective capability against ischemic injury was positively related to the expression level of nmdaR 3A on rat hippocampus slices.NR3A overexpressed cells showed higher viability than wild-type cells after cell injury.The mechanism of NR3A anti-injury ability may mainly involve in lowering intracellular Ca2+,and suppressing the generation of hydroxyl radical and NO.

关键词： nmda receptor NR3A brain hypoxia ischemia oxygen and glucose deprivation glutamate calcium overloading

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