谷氨酸及其受体与运动

谷氨酸是中枢神经系统中主要的兴奋性神经递质，它的变化与运动性中枢疲劳的产生有密切的联系．谷氨酸是通过谷氨酸受体传递兴奋性信息的，谷氨酸受体是中枢神经系统主要的兴奋性神经受体，分为离子型受体与代谢型受体．谷氨酸受体在学习．记忆的形成、脑缺氧．缺血导致的病变及其它神经系统功能中扮演着重要的角色．     

基因工程在选育高产谷氨酸菌种中的应用

本文综述了基因工程在选育谷氨酸菌种中的应用，受体、载体和供体系统的建立以及国内外科学家对谷氨酸棒杆菌所进行的分子生物学方面的研究．     

尼莫地平和银杏叶提取物对谷氨酸神经毒性的对抗作用：形态…

用形态学的方法观察了尼莫地平和银杏叶提取物对谷氨酸神经毒性的影响。结果发现谷氨酸导致新生小鼠下丘脑弓状核变性，坏，钙通道拮抗剂尼莫地平及血小板激活因子受体拮抗剂银杏叶提取物均可剂量依赖性地抑制谷氨酸引起的神经元损伤。     

尼莫地平和银杏叶提取物对谷氨酸神经毒性的对抗作用：形态学观察

用形态学的方法观察了尼莫地平和银杏叶提取物（含黄酮甙类２４％和内酯３．４％）对谷氨酸神经毒性的影响。结果发现谷氨酸导致新生小鼠下丘脑弓状核变性、坏死，钙通道拮抗剂尼莫地平及血小板激活因子受体拮抗剂银杏叶提取物均可剂量依赖性地抑制谷氨酸引起的神经元损伤。结果提示钙离子超载及血小板激活因子的激活与谷氨酸的神经毒性作用有关     

生长异质大西洋鲑转录组学分析

为探明大西洋鲑在养殖群体中存在生长异质的原因,本研究对大西洋鲑进行转录组学分析,包括生长慢速组(平均体重0.51 kg)和生长快速组(平均体重4.20 kg)各3尾鱼。结果显示,共有332 003个基因得到了注释,并筛选出948个差异表达基因,包括363个上调基因和585个下调基因。随后对差异表达基因进行GO功能富集和KEGG通路富集分析,结果表明,有543个差异表达基因被富集到61个GO条目,其中与单生物过程、细胞、结合分子功能等相关的基因较多。KEGG富集显示,共有229个差异表达基因被富集到114个代谢通路中,其中富集在钙信号通路、神经活性受体-配体交互作用信号通路、紧密连接信号通路的基因较多。通过转录组测序及功能富集分析发现,肌球蛋白、过氧化物酶体增殖物激活受体以及苏氨酸的高表达,钙/钙调蛋白依赖性蛋白激酶II、谷氨酸受体基因的低表达引起相关代谢通路调控水平的改变可能是导致大西洋鲑生长异质性的重要因素。本研究为进一步了解生长异质大西洋鲑的生物学特性、基因分子机制、代谢途径和遗传繁育提供参考依据。     

NMDA受体概述及其在学习记忆中的作用

NMDA受体是由多亚基构成的异聚体,是最重要的谷氨酸受体之一,主要分布在中枢系统中.NMDA受体有多个调节位点,能被谷氨酸、甘氨酸、Ca2+、酶等多种因素影响.NMDA受体介导的Ca2+内流广泛地参与多种生理作用,其引发的长时程增强作用而对学习记忆有极其重要的作用.     

胍基丁胺对谷氨酸诱导PC12细胞DNA损伤的影响

探讨了胍基丁胺对谷氨酸诱导PC12细胞DNA损伤的影响。在用一定浓度的谷氨酸诱导PC12细胞DNA断裂的同时，观察胍基丁胺对其DNA断裂和线粒体功能的影响，发现胍基丁胺能明显减轻谷氨酸诱导PC12细胞DNA断裂并能改善线粒体功能。这说明胍基丁胺具有抑制PC12细胞DNA损伤的作用，其机制可能与其拮抗NMDA受体，改善线粒体功能和减轻谷氨酸的氧化毒性有关。     

5-HT_(2C)受体对帕金森病模型大鼠外侧缰核谷氨酸能神经元电活动的影响

为探讨帕金森病(PD)模型大鼠外侧缰核(LHb)中谷氨酸能神经元的电活动变化及其对5-羟色胺2C(5-HT_(2C))受体刺激的反应,阐明LHb和5-HT_(2C)受体在PD神经生物学机制中发挥的作用,通过6-羟多巴胺(6-OHDA)损毁单侧黑质致密部(substantia nigra pars compacta,SNc)建立PD模型,采用细胞外记录的方法,观察假手术组与单侧SNc损毁组大鼠LHb中谷氨酸能神经元的电活动。结果显示:与假手术组大鼠相比,损毁组大鼠LHb中谷氨酸能神经元的电活动增强,表现为爆发式放电活动增强(P0.05);LHb中局部注射5-HT_(2C)受体激动剂Ro60-0175后,两组大鼠的神经元放电频率虽均升高(P0.05),但损毁组大鼠LHb中谷氨酸能神经元放电频率升高持续时间显著长于假手术组,且给药前后神经元的放电形式更趋向于爆发式活动(P0.05),变异系数明显升高(P0.05)。结果提示单侧SNc损毁使LHb中谷氨酸能神经元爆发式电活动增强,可能与PD相关抑郁行为发生有关,5-HT_(2C)受体参与了对LHb中谷氨酸能神经元电活动的调节。     

刊中刊·资讯

了解从封闭构形向活跃和钝化构形转变的结构基础，对于解读离子移变谷氨酸盐受体（NMDA受体、AMPA受体、δ受体和kainate受体）作为中枢神经系统中激发性突触传输之介质的功能来说至关重要。在受体的细胞外表面上发生的配体结合将阳离了‘选择孔打开，     

增强谷氨酸棒状杆菌羧化途径对有机酸产量的影响

以谷氨酸棒状杆菌CZ04为出发菌株,利用pK19mobsacB载体的反向筛选,通过在ppc和pyc基因前面敲入超氧化物歧化酶基因启动子(Psod),相应获得磷酸烯醇式丙酮酸羧化酶(PPC)上调的重组菌株CZ05和丙酮酸羧化酶(PYC)上调的重组菌株CZ06以及PPC、PYC双上调的重组菌株CZ07,并研究了增强谷氨酸棒状杆菌羧化途径对有机酸产量的影响。结果表明,强启动子的敲入,对菌株生长没有明显影响,但增强了谷氨酸棒状杆菌的羧化代谢途径,能促进转基因菌株中有机酸的生成和积累。     

Cloning of a cDNA for a glutamate receptor subunit activated by kainate but not AMPA.

Fast excitatory transmission in the vertebrate central nervous system is mediated mainly by L-glutamate. On the basis of pharmacological, physiological and agonist binding properties, the ionotropic glutamate receptors are classified into NMDA (N-methyl-D-aspartate), AMPA (alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionate) and kainate subtypes. Sequence homology between complementary DNA clones encoding non-NMDA glutamate receptor subunits reveals at least two subunit classes: the GluR1 to GluR4 class and the GluR5 class. Here we report the cloning and expression of a functional rat glutamate receptor subunit cDNA, GluR6, which has a very different pharmacology from that of the GluR1-GluR4 class. Receptors generated from the GluR1-GluR4 class have a higher apparent affinity for AMPA than for kainate. When expressed in Xenopus oocytes the homomeric GluR6 receptor is activated by kainate, quisqualate and L-glutamate but not by AMPA, and the apparent affinity for kainate is higher than for receptors from the GluR1-GluR4 class. Desensitization of the receptor was observed with continuous application of agonist. The homomeric GluR6 glutamate receptor exhibits an outwardly rectifying current-voltage relationship. In situ hybridizations reveal a pattern of GluR6 gene expression reminiscent of the binding pattern obtained with [3H]kainate.     

Pertussis toxin reverses adenosine inhibition of neuronal glutamate release

Adenosine and its analogues are potent inhibitors of synaptic activity in the central and peripheral nervous system. In the central nervous system (CNS), this appears to arise primarily by inhibition of presynaptic release of transmitters, including glutamate, which is possibly the major excitatory transmitter in the brain. In addition, postsynaptic effects of adenosine have been reported which would also serve to reduce neurotransmission. The mechanism by which adenosine inhibits CNS neurotransmission is unknown, although it appears to exert its effect via an A1 receptor which in some systems is negatively coupled to adenylate cyclase. In an attempt to elucidate the mechanism of inhibition, we have examined the effect of pertussis toxin (PTX) on the ability of the stable adenosine analogue (-)phenylisopropyladenosine (PIA) to inhibit glutamate release from cerebellar neurones maintained in primary culture. PTX, by ADP-ribosylating the nucleotide-binding protein Ni, prevents coupling of inhibitory receptors such as the A1 receptor to adenylate cyclase. As reported here, we found that PTX, as well as preventing inhibition of adenylate cyclase by PIA, also converts the PIA-induced inhibition of glutamate release to a stimulation. Our results suggest strongly that purinergic inhibitory modulation of transmitter release occurs by inhibition of adenylate cyclase.     

Suppression by glutamate of cGMP-activated conductance in retinal bipolar cells.

Depolarizing bipolar cells (DBCs) of the retina are the only neurons in the vertebrate central nervous system known to be hyperpolarized by the neurotransmitter glutamate. Both glutamate and its analogue L-2-amino-4-phosphonobutyrate (APB) hyperpolarize DBCs by decreasing membrane conductance. Furthermore, glutamate responses in DBCs slowly decrease during whole-cell recording, suggesting that the response involves a second messenger system. Here we report that intracellular cyclic GMP or GTP activates a membrane conductance that is suppressed by APB, resulting in an enhanced APB response. In the presence of GTP-gamma-S, APB causes an irreversible suppression of the conductance. Inhibitors of G-protein activation or phosphodiesterase activity decrease the APB response. Thus, the DBC glutamate receptor seems to close ion channels by increasing the rate of cGMP hydrolysis by a G protein-mediated process that is strikingly similar to light transduction in photoreceptors.     

Multiple-conductance channels activated by excitatory amino acids in cerebellar neurons

In the mammalian central nervous system amino acids such as L-glutamate and L-aspartate are thought to act as fast synaptic transmitters. It has been suggested that at least three pharmacologically-distinguishable types of glutamate receptor occur in central neurons and that these are selectively activated by the glutamate analogues N-methyl-D-aspartate (NMDA), quisqualate and kainate. These three receptor types would be expected to open ion channels with different conductances. Hence if agonists produce similar channel conductances this would suggest they are acting on the same receptor. Another possibility is suggested by experiments on spinal neurons, where GABA (gamma-amino butyric acid) and glycine appear to open different sub-conductance levels of one class of channel while acting on different receptors. By analogy, several types of glutamate receptor could also be linked to a single type of channel with several sub-conductance states. We have examined these possibilities in cerebellar neurons by analysing the single-channel currents activated by L-glutamate, L-aspartate, NMDA, quisqualate and kainate in excised membrane patches. All of these agonists are capable of opening channels with at least five different conductance levels, the largest being about 45-50 pS. NMDA predominantly activated conductance levels above 30 pS while quisqualate and kainate mainly activated ones below 20 pS. The presence of clear transitions between levels favours the idea that the five main levels are all sub-states of the same type of channel.     

Structural basis of glutamate recognition by a dimeric metabotropic glutamate receptor

The metabotropic glutamate receptors (mGluRs) are key receptors in the modulation of excitatory synaptic transmission in the central nervous system. Here we have determined three different crystal structures of the extracellular ligand-binding region of mGluR1--in a complex with glutamate and in two unliganded forms. They all showed disulphide-linked homodimers, whose 'active' and 'resting' conformations are modulated through the dimeric interface by a packed alpha-helical structure. The bi-lobed protomer architectures flexibly change their domain arrangements to form an 'open' or 'closed' conformation. The structures imply that glutamate binding stabilizes both the 'active' dimer and the 'closed' protomer in dynamic equilibrium. Movements of the four domains in the dimer are likely to affect the separation of the transmembrane and intracellular regions, and thereby activate the receptor. This scheme in the initial receptor activation could be applied generally to G-protein-coupled neurotransmitter receptors that possess extracellular ligand-binding sites.     

Glycine binding primes NMDA receptor internalization

NMDA (N-methyl-d-aspartate) receptors (NMDARs) are a principal subtype of excitatory ligand-gated ion channel with prominent roles in physiological and disease processes in the central nervous system. Recognition that glycine potentiates NMDAR-mediated currents as well as being a requisite co-agonist of the NMDAR subtype of 'glutamate' receptor profoundly changed our understanding of chemical synaptic communication in the central nervous system. The binding of both glycine and glutamate is necessary to cause opening of the NMDAR conductance pore. Although binding of either agonist alone is insufficient to cause current flow through the channel, we report here that stimulation of the glycine site initiates signalling through the NMDAR complex, priming the receptors for clathrin-dependent endocytosis. Glycine binding alone does not cause the receptor to be endocytosed; this requires both glycine and glutamate site activation of NMDARs. The priming effect of glycine is mimicked by the NMDAR glycine site agonist d-serine, and is blocked by competitive glycine site antagonists. Synaptic as well as extrasynaptic NMDARs are primed for internalization by glycine site stimulation. Our results demonstrate transmembrane signal transduction through activating the glycine site of NMDARs, and elucidate a model for modulating cell-cell communication in the central nervous system.     

Subunit arrangement and function in NMDA receptors

Excitatory neurotransmission mediated by NMDA (N-methyl-D-aspartate) receptors is fundamental to the physiology of the mammalian central nervous system. These receptors are heteromeric ion channels that for activation require binding of glycine and glutamate to the NR1 and NR2 subunits, respectively. NMDA receptor function is characterized by slow channel opening and deactivation, and the resulting influx of cations initiates signal transduction cascades that are crucial to higher functions including learning and memory. Here we report crystal structures of the ligand-binding core of NR2A with glutamate and that of the NR1-NR2A heterodimer with glutamate and glycine. The NR2A-glutamate complex defines the determinants of glutamate and NMDA recognition, and the NR1-NR2A heterodimer suggests a mechanism for ligand-induced ion channel opening. Analysis of the heterodimer interface, together with biochemical and electrophysiological experiments, confirms that the NR1-NR2A heterodimer is the functional unit in tetrameric NMDA receptors and that tyrosine 535 of NR1, located in the subunit interface, modulates the rate of ion channel deactivation.     

Cloning of a putative high-affinity kainate receptor expressed predominantly in hippocampal CA3 cells.

Kainic acid is a potent neurotoxin for certain neurons. Its neurotoxicity is thought to be mediated by an excitatory amino-acid-gated ion channel (ionotropic receptor) possessing nanomolar affinity for kainate. Here we describe a new member of the rat excitatory amino-acid receptor gene family, KA-1, that has a 30% sequence similarity with the previously characterized alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor subunits GluR-A to -D. The pharmacological profile of expressed recombinant KA-1 determined in binding experiments with [3H]kainate is different from that of the cloned AMPA receptors and similar to the mammalian high-affinity kainate receptor (kainate greater than quisqualate greater than glutamate much greater than AMPA) with a dissociation constant of about 5 nM for kainate. The selectively high expression of KA-1 messenger RNA in the CA3 region of the hippocampus closely corresponds to autoradiographically located high-affinity kainate binding sites. This correlation, as well as the particular in vivo pattern of neurodegeneration observed on kainate-induced neurotoxicity, suggests that KA-1 participates in receptors mediating the kainate sensitivity of neurons in the central nervous system.     

Glutamate release in severe brain ischaemia is mainly by reversed uptake

The release of glutamate during brain anoxia or ischaemia triggers the death of neurons, causing mental or physical handicap. The mechanism of glutamate release is controversial, however. Four release mechanisms have been postulated: vesicular release dependent on external calcium or Ca2+ released from intracellular stores; release through swelling-activated anion channels; an indomethacin-sensitive process in astrocytes; and reversed operation of glutamate transporters. Here we have mimicked severe ischaemia in hippocampal slices and monitored glutamate release as a receptor-gated current in the CA1 pyramidal cells that are killed preferentially in ischaemic hippocampus. Using blockers of the different release mechanisms, we demonstrate that glutamate release is largely by reversed operation of neuronal glutamate transporters, and that it plays a key role in generating the anoxic depolarization that abolishes information processing in the central nervous system a few minutes after the start of ischaemia. A mathematical model incorporating K+ channels, reversible uptake carriers and NMDA (N-methyl-D-aspartate) receptor channels reproduces the main features of the response to ischaemia. Thus, transporter-mediated glutamate homeostasis fails dramatically in ischaemia: instead of removing extracellular glutamate to protect neurons, transporters release glutamate, triggering neuronal death.