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Attenuation of nMdA receptor activity and neurotoxicity by nitroxyl anion, nO^-

nEUROn 1999年第2期24卷 461-469页

作者： Kim, WK Choi, YB Rayudu, PV das, P Asaad, W Arnelle, dR Stamler, JS Lipton, SA Brigham & Womens Hosp Cerebrovasc & Neurosci Res Inst Boston MA 02115 USA Harvard Univ Sch Med Program Neurosci Boston MA 02115 USA Duke Univ Med Ctr Howard Hughes Med Inst Dept Med Durham NC 27710 USA Duke Univ Med Ctr Dept Cell Biol Durham NC 27710 USA

Recent evidence indicates that the nO-related species, nitroxyl anion (nO-), is produced in physiological systems by several redox metal-containing proteins, including hemoglobin, nitric oxide synthase (nOS), superoxide dismutase, and S-nitrosothiols (SnOs), which have recently been identified in brain. However, the chemical biology of nO- remains largely unknown. Here, we show that nO--unlike nO., but reminiscent of nO+ transfer (or S-nitrosylation) - reacts mainly with Cys-399 in the nR2A subunit of the n-methyl-d-aspartate (nMdA) receptor to curtail excessive Ca2+ influx and thus provide neuroprotection from excitotoxic insults. This effect of nO- closely resembles that of nOS, which also downregulates nMdA receptor activity under similar conditions in culture.

关键词：阴离子/代谢细胞培养的大脑皮质/细胞学 n-甲基天冬氨酸/药理学神经元/代谢神经毒素类/代谢一氧化氮/药理学一氧化氮供体/药理学一氧化氮合酶/代谢氧化氮类/代谢卵母细胞大鼠 Sprague-dawley 受体 n-甲基-d-天冬氨酸/化学受体 n-甲基-d-天冬氨酸/药物作用受体 n-甲基-d-天冬氨酸/代谢受体 n-甲基-d-天冬氨酸/生理学巯基化合物/化学巯基化合物/代谢光滑爪蟾动物女(雌)性大鼠

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Early postnatal switch in magnesium sensitivity of nMdA receptors in rat CA1 pyramidal cells

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JOURnAL OF PHYSIOLOGY-LOndOn 1999年第1期521卷 99-111页

作者： Kirson, Ed Schirra, C Konnerth, A Yaari, Y Hebrew Univ Jerusalem Hadassah Fac Med Inst Med Sci Dept Physiol IL-91120 Jerusalem Israel Univ Saarland Inst Physiol 1 D-66421 Homburg Germany

1. Whole-cell patch-damp recordings of iontophoretically induced n-methyl-d-aspartate (nMdA) receptor-mediated currents (I-nMdA) in CA1 pyramidal cells in hippocampal slices from 1- to 40-day-old rats were used to characterize developmental changes in the Mg2+ sensitivity of nMdA receptors. 2. The dose-response relations for extracellular Mg2+ blockade of I-nMdA indicated a high affinity binding of Mg2+ to nMdA receptors at membrane potentials more negative than -60 mV, independent of postnatal age. 3. depolarizing the cells unblocked nMdA receptors by decreasing their affinity for Mg2+. The efficacy of depolarization in unblocking nMdA receptors markedly increased after postnatal day 4 (P4), endowing the receptors with a greater voltage dependence. 4. The nR2B subunit-specific nMdA antagonist ifenprodil reduced I-nMdA in pyramidal cells of all ages. The sensitivity of I-nMdA to ifenprodil was greatest during the first postnatal week and decreased thereafter, indicating an enhanced contribution of nR2B subunit-containing nMdA receptors to I-nMdA in the first week after birth. 5. In the first postnatal week, the ifenprodil-insensitive InMdA component had a lower voltage dependence than the total I-nMdA. In older pyramidal cells, the voltage dependence of the ifenprodil-insensitive component and the total I-nMdA were similar. 6. In another set of CA1 pyramidal cells, single-cell reverse transcription and polymerase chain reaction (RT-PCR) were used to characterize concomitant developmental changes in nMdA subunit mRnA expression. The mRnA for the nR2d subunit was detected during the first postnatal week in 50% of the cells and disappeared thereafter. The proportion of cells expressing the nR2A and nR2B subunits remained relatively constant throughout the first five postnatal weeks. 7. We conclude that nMdA receptors in hippocampal CA1 pyramidal cells are effectively blocked by Mg2+ at all ages. After 4 days they become much less sensitive to Mg2+ at depolarized membra

关键词：动物新生基因表达调控发育期海马/细胞学海马/生长和发育海马/代谢镁/药理学膜电位膜片钳术哌啶类/药理学蛋白质结构四级锥体细胞/药物作用锥体细胞/代谢 RnA 信使/遗传学 RnA 信使/代谢大鼠 Wistar 受体 n-甲基-d-天冬氨酸/药物作用受体 n-甲基-d-天冬氨酸/遗传学受体 n-甲基-d-天冬氨酸/代谢逆转录聚合酶链反应动物大鼠

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d-serine is an endogenous ligand for the glycine site of the n-methyl-d-aspartate receptor

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PROCEEdInGS OF THE nATIOnAL ACAdEMY OF SCIEnCES OF THE UnITEd STATES OF AMERICA 2000年第9期97卷 4926-4931页

作者： Mothet, JP Parent, AT Wolosker, H Brady, RO Linden, dJ Ferris, Cd Rogawski, MA Snyder, SH Johns Hopkins Univ Sch Med Dept Neurosci Baltimore MD 21205 USA Johns Hopkins Univ Sch Med Dept Pharmacol & Mol Sci Baltimore MD 21205 USA Johns Hopkins Univ Sch Med Dept Psychiat Baltimore MD 21205 USA NINDS Epilepsy Res Branch NIH Bethesda MD 20892 USA

Functional activity of n-methyl-d-aspartate (nMdA) receptors requires both glutamate binding and the binding of an endogenous coagonist that has been presumed to be glycine, although d-serine is a more potent agonist, Localizations of d-serine and it biosynthetic enzyme serine racemase approximate the distribution of nMdA receptors more closely than glycine, We now show that selective degradation of d-serine with d-amino acid oxidase greatly attenuates nMdA receptor-mediated neurotransmission as assessed by using whole-cell patch-clamp recordings or indirectly by using biochemical assays of the sequelae of nMdA receptor-mediated calcium flux. The inhibitory effects of the enzyme are fully reversed by exogenously applied d-serine, which by itself did not potentiate nMdA receptor-mediated synaptic responses. Thus, d-serine is an endogenous modulator of the glycine site of nMdA receptors and fully occupies this site at some functional synapses.

关键词：衰老星形细胞/生理学细胞培养的小脑/生长和发育小脑/生理学 d-氨基酸氧化酶/代谢胚胎哺乳动物甘氨酸海马/生理学配体神经元/生理学喹恶啉类/药理学大鼠 Sprague-dawley 受体 n-甲基-d-天冬氨酸/化学受体 n-甲基-d-天冬氨酸/药物作用受体 n-甲基-d-天冬氨酸/生理学丝氨酸/代谢立体异构现象动物男(雄)性大鼠

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dynorphin A elicits an increase in intracellular calcium in cultured neurons via a non-opioid, non-nMdA mechanism

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JOURnAL OF nEUROPHYSIOLOGY 2000年第5期83卷 2610-2615页

作者： Tang, QB Lynch, RM Porreca, F Lai, J Univ Arizona Hlth Sci Ctr Dept Pharmacol Tucson AZ 85724 USA Univ Arizona Hlth Sci Ctr Dept Physiol Tucson AZ 85724 USA

The opioid peptide dynorphin A is known to elicit a number of pathological effects that may result from neuronal excitotoxicity. An up-regulation of this peptide has also been causally related to the dysesthesia associated with inflammation and nerve injury. These effects of dynorphin A are not mediated through opioid receptor activation but can be effectively blocked by pretreatment with n-methyl-d-aspartate (nMdA) receptor antagonists, thus implicating the excitatory amino acid system as a mediator of the actions of dynorphin A and/or its fragments. A direct interaction between dynorphin A and the nMdA receptors has been well established;however the physiological relevance of this interaction remains equivocal. This study examined whether dynorphin A elicits a neuronal excitatory effect that may underlie its activation of the nMdA receptors. Calcium imaging of individual cultured cortical neurons showed that the nonopioid peptide dynorphin A(2-17) induced a time- and dose-dependent increase in intracellular calcium. This excitatory effect of dynorphin A(2-17) was insensitive to (+)- 5-methyl-10,11-dihydro-5H-dibenzo[a, d]-cyclohepten-5,10-imine (MK-801) pretreatment in nMdA-responsive cells. Thus dynorphin A stimulates neuronal cells via a nonopioid, non-nMdA mechanism. This excitatory action of dynorphin A could modulate nMdA receptor activity in vivo by enhancing excitatory neurotransmitter release or by potentiating nMdA receptor function in a calcium-dependent manner. Further characterization of this novel site of action of dynorphin A may provide new insight into the underlying mechanisms of dynorphin excitotoxicity and its pathological role in neuropathy.

关键词：钙/代谢细胞培养的大脑皮质/细胞学大脑皮质/药物作用地卓西平马来酸盐/药理学剂量效应关系药物药物协同作用强啡肽类/药理学兴奋性氨基酸拮抗剂/药理学细胞内液/药物作用细胞内液/代谢 n-甲基天冬氨酸/药理学神经元/细胞学神经元/药物作用神经元/代谢肽碎片/药理学受体 n-甲基-d-天冬氨酸/药物作用受体阿片样/药物作用动物女(雌)性男(雄)性大鼠

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n-methyl-d-aspartate (nMdA) receptor: friend or foe?

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Indian Journal of Experimental Biology 1998年第10期36卷 947-959页

作者： Robichon, Regine Ticku, Maharaj K. Department of Pharmacology University of Texas Health Science Center at San Antonio 78284-7764 USA.

Glutamate is a nonessential amino acid involved in biochemical reactions such as detoxification of ammonia in CnS, synthesis of proteins, and a precursor for the neurotransmitter, GABA. It is also an excitatory amino acid neurotransmitter so-called glutamate receptors, however, it can also be a powerful neurotoxin. Over the years, specific agonists and antagonists have been designed to distinguish between various glutamate receptors. So far, these receptors are comprised ionotropic receptors such as n-methyl-d-aspartate (nMdA), α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) and kainate, and metabotropic receptors (mGluR) and L-AP4. Of these receptor subtypes, nMdA receptors have received a great deal of attention because of their involvement in both physiological and pathological processes in CnS. Thus, it is important to understand how receptor works as a protein complex in order to answer the question whether it is an enemy or a friend. In this review, we will describe its functions, composition, and inter-relationship of various binding sites associated with this receptor protein complex.

关键词：受体谷氨酸/药物作用受体谷氨酸/生理学受体 n-甲基-d-天冬氨酸/药物作用受体 n-甲基-d-天冬氨酸/生理学构效关系动物人类

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Zinc and ifenprodil allosterically inhibit two separate polyamine-sensitive sites at n-methyl-d-aspartate receptor complex

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JOURnAL OF PHARMACOLOGY And EXPERIMEnTAL THERAPEUTICS 1999年第3期289卷 1584-1591页

作者： Berger, ML Rebernik, P Univ Vienna Inst Biochem Pharmacol A-1090 Vienna Austria

In this study, we investigated the hypothesis that inhibition of the n-methyl-d-aspartate (nMdA) receptor complex by zinc involves a polyamine-sensitive regulatory site. We found that the specific binding of the open channel ligand [H-3]MK-801 to rat hippocampal membranes 1) was inhibited by low concentrations of Zn2+ (IC50 = 5.5 mu M) by 65%. 2) This high-affinity component of inhibition was reversed by the polyamine spermine to an extent that could be reconciled with competitive interaction between Zn2+ and spermine. 3) Partial inhibition by Zn2+ was additive with partial inhibition by ifenprodil, an inhibitor of the nMdA receptor complex supposed to act at a polyamine-sensitive regulatory site, and 4) in membranes prepared from several other brain regions, inhibition of [H-3]MK-801 binding by Zn2+ and by ifenprodil was either less than additive, or superadditive. Our observation that ifenprodil, at concentrations saturating its high-affinity component of inhibition, prevented spermine from reversing the inhibition by Zn2+ indicates that spermine did not increase [H-3]MK-801 binding by competition with Zn2+ but rather via another polyamine regulatory site not sensitive to zinc but sensitive to ifenprodil. We conclude that Zn2+ reduces channel opening of the nMdA receptor complex by allosteric inhibition of a polyamine-sensitive regulatory site different from that inhibited by ifenprodil and that these two allosteric sites influence each other in a manner dependent on the brain region investigated. The different proportions of zinc/ifenprodil inhibition in different regions could reflect different percentages of various nMdA receptor subtypes.

关键词：别构调节结合竞争性脑/代谢细胞膜/代谢齿状回/代谢地卓西平马来酸盐/药代动力学兴奋性氨基酸拮抗剂/药理学海马/代谢动力学神经元/代谢器官特异性哌啶类/药理学放射配体测定大鼠 Wistar 受体 n-甲基-d-天冬氨酸/化学受体 n-甲基-d-天冬氨酸/药物作用受体 n-甲基-d-天冬氨酸/代谢精胺/药理学氚锌/药理学动物男(雄)性大鼠

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Geometry and charge determine pharmacological effects of steroids on n-methyl-d-aspartate receptor-induced Ca²⁺ accumulation and cell death

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JOURnAL OF PHARMACOLOGY And EXPERIMEnTAL THERAPEUTICS 2000年第3期293卷 747-754页

作者： Weaver, CE Land, MB Purdy, RH Richards, KG Gibbs, TT Farb, dH Boston Univ Sch Med Dept Pharmacol Mol Neurobiol Lab Boston MA 02118 USA

Modulation of n-methyl-d-aspartate (nMdA) receptor function by a series of sulfated steroids and dicarboxylic acid ester analogs of pregnenolone sulfate and pregnanolone sulfate was investigated in cultured hippocampal neurons. The "bent" steroid ring structure associated with 5 beta-stereochemistry favors receptor inhibition, whereas the more planar ring structure of the pregn-5-enes and 5 alpha-pregnanes favors potentiation of nMdA-induced [Ca2+] increases and neuronal cell death. The nature of the negatively charged group attached to the steroid C3 position is important for both the neuroprotection afforded by pregnane steroids and the exacerbation of nMdA-induced neuronal death by pregn-5-enes. dicarboxylic acid hemiesters of various lengths can substitute for the sulfate group of the positive modulator pregnenolone sulfate and the negative modulator pregnanolone sulfate. This result suggests that precise coordination with the oxygen atoms of the sulfate group is not critical for modulation and that the steroid recognition sites can accommodate bulky substituents at C3. The capacity of charged steroids to enhance or protect against nMdA-induced death of hippocampal neurons is strongly correlated with modulation of nMdA-induced Ca2+ accumulation, indicating that direct enhancement or inhibition of nMdA receptor function is responsible for the proexcitotoxic or neuroprotective effects of these steroids.

关键词：钙/代谢细胞死亡/药物作用海马/药物作用分子构象 n-甲基天冬氨酸/毒性大鼠 Sprague-dawley 受体 n-甲基-d-天冬氨酸/药物作用受体 n-甲基-d-天冬氨酸/生理学甾类/药理学构效关系动物大鼠

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Modulation of neuronal and recombinant GABA_A receptors by redox reagents

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JOURnAL OF PHYSIOLOGY-LOndOn 1999年第1期517卷 35-50页

作者： Amato, A Connolly, Cn Moss, SJ Smart, TG Univ London Sch Pharm Dept Pharmacol London WC1N 1AX England UCL MRC Inst Mol Cell Biol London WC1E 6BT England UCL Dept Pharmacol London WC1E 6BT England

1. The functional role played by the postulated disulphide bridge in gamma-aminobutyric acid type A (GABA(A)) receptors and its susceptibility to oxidation and reduction were studied using recombinant (murine receptor subunits expressed in human embryonic kidney cells) and rat neuronal GABA(A) receptors in conjunction with whole-cell and single channel patch-clamp techniques. 2. The reducing agent dithiothreitol (dTT) reversibly potentiated GABA-activated responses (I(GABA)) of alpha 1 beta 1 or alpha 1 beta 2 receptors while the oxidizing reagent 5,5'-dithio-bis-(2-nitrobenzoic acid) (dTnB) caused inhibition. Redox modulation of I(GABA) was independent of GABA concentration, membrane potential and the receptor agonist and did not affect the GABA EC(50) or Hill coefficient. The endogenous antioxidant reduced glutathione (GSH) also potentiated I(GABA) in alpha 1 beta 2 receptors, while both the oxidized form of dTT and glutathione (GSSG) caused small inhibitory effects. 3. Recombinant receptors composed of alpha 1 beta 1 gamma 2S or alpha 1 beta 2 gamma 2S were considerably less sensitive to dTT and dTnB. 4. For neuronal GABA(A) receptors, I(GABA) was enhanced flurazepam and relatively unaffected by redox reagents. However, in cultured sympathetic neurones, nicotinic acetylcholine-activated responses were inhibited by dTT whilst in cerebellar granule neurones, nMdA-activated currents were potentiated by dTT and inhibited by: dTnB. 5. Single GABA-activated ion channel currents exhibited a conductance of 16 pS for alpha 1 beta 1 constructs. dTT did not affect the conductance or individual open time constants determined from dwell time histograms, but increased the mean open time by affecting the channel open probability without increasing the number of cell surface receptors. 6. A kinetic model of the effects of dTT and dTnB suggested that the receptor existed in equilibrium between oxidized and reduced forms. dTT increased the rate of entry into reduced receptor forms

关键词：细胞系细胞培养的二硫硝基苯甲酸/药理学二硫苏糖醇/药理学动力学膜电位模型生物学神经元/药物作用神经元/代谢氧化还原膜片钳术蛋白质构象受体 GABA-A/药物作用受体 GABA-A/遗传学受体 GABA-A/代谢受体 n-甲基-d-天冬氨酸/药物作用受体 n-甲基-d-天冬氨酸/代谢受体烟碱/药物作用受体烟碱/代谢重组蛋白质类/化学重组蛋白质类/遗传学重组蛋白质类/代谢巯基试剂/药理学动物人类小鼠大鼠

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Wind-up of spinal cord neurones and pain sensation: much ado about something?

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PROGRESS In nEUROBIOLOGY 2000年第2期61卷 169-203页

作者： Herrero, JF Laird, JMA Lopez-Garcia, JA Univ Alcala de Henares Dept Fisiol Madrid 28871 Spain

Wind-up is a frequency-dependent increase in the excitability of spinal cord neurones, evoked by electrical stimulation of afferent C-fibres. Although it has been studied over the past thirty years, there are still uncertainties about its physiological meaning. Glutamate (nMdA) and tachykinin nK1 receptors are required to generate wind-up and therefore a positive modulation between these two receptor types has been suggested by some authors. However, most drugs capable of reducing the excitability of spinal cord neurones, including opioids and nSAIds, can also reduce or even abolish wind-up. Thus, other theories involving synaptic efficacy, potassium channels, calcium channels, etc, have also been proposed for the generation of this phenomenon. Whatever the mechanisms involved in its generation, wind-up has been interpreted as a system for the amplification in the spinal cord of the nociceptive message that arrives from peripheral nociceptors connected to C-fibres. This probably reflects the physiological system activated in the spinal cord after an intense or persistent barrage of afferent nociceptive impulses. On the other hand, wind-up, central sensitisation and hyperalgesia are not the same phenomena, although they may share common properties. Wind-up can be an important tool to study the processing of nociceptive information in the spinal cord, and the central effects of drugs that modulate the nociceptive system. This paper reviews the physiological and pharmacological data on wind-up of spinal cord neurones, and the perceptual correlates of wind-up in human subjects, in the context of its possible relation to the triggering of hyperalgesic stales, and also the multiple factors which contribute to the generation of wind-up. (C) 2000 Elsevier Science Ltd. All rights reserved.

关键词：传入通路/生理学镇痛药/药理学麻醉药局部/药理学消炎药非甾类/药理学痛觉过敏/病理生理学炎症离子转运/药物作用模型神经学吗啡/药理学神经纤维/药物作用神经纤维/生理学神经组织蛋白质类/药物作用神经组织蛋白质类/生理学神经痛/病理生理学神经元可塑性/药物作用神经元可塑性/生理学神经元传入/药物作用神经元传入/生理学神经肽类/生理学疼痛感受器/生理学疼痛/药物疗法疼痛/病理生理学受体亲代谢性谷氨酸盐/药物作用受体亲代谢性谷氨酸盐/生理学受体 n-甲基-d-天冬氨酸/药物作用受体 n-甲基-d-天冬氨酸/生理学受体神经激肽1/药物作用受体神经激肽1/生理学反射/生理学脊髓/细胞学脊髓/病理生理学 P物质/生理学突触传递/药物作用内脏/神经支配动物猫人类

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Regional distribution and pharmacological characteristics of [³H]n-acetyl-aspartyl-glutamate (nAAG) binding sites in rat brain

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nEUROCHEMISTRY InTERnATIOnAL 2001年第1期38卷 53-62页

作者： Shave, E Pliss, L Lawrance, ML FitzGibbon, T Stastny, F Balcar, VJ Univ Sydney Dept Anat & Histol Sydney NSW 2006 Australia Univ Sydney Inst Biomed Res Sydney NSW 2006 Australia Acad Sci Prague Inst Physiol Dept Mol Neurobiol CZ-14220 Prague Czech Republic

Autoradiographical studies revealed that 10 nM [H-3]n-acetyl-aspartyl-glutamate (nAAG) labelled grey matter structures, particularly in the hippocamus, cerebral neocortex, striatum, septal nuclei and the cerebellar cortex. The binding was inhibited by (2S,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl)-glycine (dCG IV), an agonist at group II metabotropic glutamate receptors (mGluR II). (RS)-alpha-Methyl-4-tetrazolylphenylglycine (MTPG), (RS)-alpha-cyclopropyl-4-phosphonoglycine (CPPG) and (RS)-alpha-methylserine-O-phosphate monophenyl ester (MSOPPE), all antagonists at mGluR II and mGluR III, also inhibited [H-3]nAAG binding. Other inhibitors were (1S,3R)-1-aminocyclopentane-1,3-dicarboxylate (ACPd), a broad-spectrum mGluR agonist with preference for groups I and II and the mGluR I agonists/mGluR II antagonists (S)-3-carboxy-4-hydroxyphenylglycine (3,4-CHPG) and (S)-4-carboxy-3-hydroxyphenylglycine (4,3-CHPG). neither the mGluR I specific agonist (S)-dihydroxyphenylglycine nor any of the ionotropic glutamate receptor ligands such as kainate, AMPA and MK-801 had strong effects (except for the competitive nMdA antagonist CGS 19755, which produced 20-40% inhibition at 100 mu M) suggesting that, at low nM concentrations, [3H]nAAG binds predominantly to metabotropic glutamate receptors, particularly those of the mGluR II type. Several studies have indicated that nAAG can interact with mGluR II and the present study supports this notion by demonstrating that sites capable of binding nAAG at low concentrations and displaying pharmacological characteristics of mGluR II exist in the central nervous tissue. Furthermore, the results show that autoradiography of [H-3]nAAG binding can be used to quantify the distribution of such sites in distinct brain regions and study their pharmacology at the same time. (C) 2000 Elsevier Science Ltd. All rights reserved.

关键词：丙氨酸/类似物和衍生物丙氨酸/药理学结合部位脑/超微结构脑化学羧肽酶类/代谢冷温度环亮氨酸/类似物和衍生物环亮氨酸/药理学环丙烷类/药理学二肽类/代谢二肽类/药理学地卓西平马来酸盐/药理学兴奋性氨基酸激动剂/药理学兴奋性氨基酸拮抗剂/药理学谷氨酸羧肽酶Ⅱ 甘氨酸/类似物和衍生物甘氨酸/药理学红藻氨酸/药理学神经组织蛋白质类/药物作用神经组织蛋白质类/代谢磷酸丝氨酸/类似物和衍生物磷酸丝氨酸/药理学哌可酸/药理学大鼠 Sprague-dawley 受体谷氨酸/药物作用受体谷氨酸/代谢受体亲代谢性谷氨酸盐/药物作用受体亲代谢性谷氨酸盐/代谢受体 n-甲基-d-天冬氨酸/药物作用受体 n-甲基-d-天冬氨酸/代谢四唑类/药理学 α-氨基-3-羟基-5-甲基-4-异唑丙酸/药理学动物女(雌)性男(雄)性大鼠

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