Role of NMDA receptor subtypes in governing the direction of hippocampal synaptic plasticity

Activation of N-methyl-d-aspartate subtype glutamate receptors (NMDARs) is required for long-term potentiation (LTP) and long-term depression (LTD) of excitatory synaptic transmission at hippocampal CA1 synapses, the proposed cellular substrates of learning and memory. However, little is known about how activation of NMDARs leads to these two opposing forms of synaptic plasticity. Using hippocampal slice preparations, we showed that selectively blocking NMDARs that contain the NR2B subunit abolishes the induction of LTD but not LTP. In contrast, preferential inhibition of NR2A-containing NMDARs prevents the induction of LTP without affecting LTD production. These results demonstrate that distinct NMDAR subunits are critical factors that determine the polarity of synaptic plasticity.     

Enhanced and delayed stress-induced alcohol drinking in mice lacking functional CRH1 receptors

There is a relation between stress and alcohol drinking. We show that the corticotropin-releasing hormone (CRH) system that mediates endocrine and behavioral responses to stress plays a role in the control of long-term alcohol drinking. In mice lacking a functional CRH1 receptor, stress leads to enhanced and progressively increasing alcohol intake. The effect of repeated stress on alcohol drinking behavior appeared with a delay and persisted throughout life. It was associated with an up-regulation of the N-methyl-d-aspartate receptor subunit NR2B. Alterations in the CRH1 receptor gene and adaptional changes in NR2B subunits may constitute a genetic risk factor for stress-induced alcohol drinking and alcoholism.     

NR2B的同源模建及其与Con-G的对接(英文)

利用3种同源蛋白结构,通过同源模建的方法构建N-甲基-D-天冬氨酸(NMDA)受体NR2B亚基的三维构象,并对模建蛋白进行多种参数的验证.结果表明:在构建的模型中,conantokin-G(Con-G,NR2B亚基受体拮抗剂)与NR2B对接后,Con-G能很好地嵌入到NR2B亚基激动剂结合部位,并且Con-G的构象基本不变,仍保持α-螺旋构象;Con-G的E2、Gla4、L5、Q9、I12和Q13,NR2B的E420、S421、D423、K458和D715是参与相互作用的重要氨基酸,它们之间形成了一些重要的氢键.该模型的建立对预测NMDA受体与其配体的相互作用具有一定的作用,为设计新的NMDA受体激动剂和拮抗剂提供了重要线索.     

Asymmetrical allocation of NMDA receptor epsilon2 subunits in hippocampal circuitry

Despite its implications for higher order functions of the brain, little is currently known about the molecular basis of left-right asymmetry of the brain. Here we report that synaptic distribution of N-methyl-D-aspartate (NMDA) receptor GluRepsilon2 (NR2B) subunits in the adult mouse hippocampus is asymmetrical between the left and right and between the apical and basal dendrites of single neurons. These asymmetrical allocations of epsilon2 subunits differentiate the properties of NMDA receptors and synaptic plasticity between the left and right hippocampus. These results provide a molecular basis for the structural and functional asymmetry of the mature brain.     

An oral vaccine against NMDAR1 with efficacy in experimental stroke and epilepsy

The brain is generally considered immunoprivileged, although increasing examples of immunological responses to brain antigens, neuronal expression of major histocompatibility class I genes, and neurological autoimmunity have been recognized. An adeno-associated virus (AAV) vaccine generated autoantibodies that targeted a specific brain protein, the NR1 subunit of the N-methyl-D-aspartate (NMDA) receptor. After peroral administration of the AAV vaccine, transgene expression persisted for at least 5 months and was associated with a robust humoral response in the absence of a significant cell-mediated response. This single-dose vaccine was associated with strong anti-epileptic and neuroprotective activity in rats for both a kainate-induced seizure model and also a middle cerebral artery occlusion stroke model at 1 to 5 months following vaccination. Thus, a vaccination strategy targeting brain proteins is feasible and may have therapeutic potential for neurological disorders.     

昼夜变温下高温与干旱胁迫对Bt棉毒蛋白含量的影响及其生理机制

【目的】明确Bt棉叶片Cry1Ac毒蛋白含量对昼夜变温下高温干旱胁迫响应及其生理机制,为生产中Bt棉抗虫性的安全稳定利用提供参考。【方法】2019—2020年在扬州大学农学院,以常规种泗抗1号（SK-1）和杂交种泗抗3号（SK-3）为材料,以温度和土壤水分含量为因子,温度分别设为34℃（白天,7:00—19:00）/28℃（夜间,19:00—7:00）(A1)、38℃/28℃（A2）;土壤水分含量分别为田间土壤最大持水量的50%(B1)和60%(B2),并以32℃/28℃、田间土壤最大持水量的75%为对照（CK）。各处理分别持续4、7、10 d(DAS)。【结果】不同处理导致叶片中Cry1Ac毒蛋白含量降低,且随着胁迫时间的延长,下降幅度增加。处理间相比,A1B2处理下降幅度最少,7 DAS后开始显著低于CK;A1B1处理下降幅度其次,4 DAS后显著低于CK;A2B1、A2B2处理在4 DAS显著下降。可溶性蛋白（SP）含量、硝酸还原酶（NR）、谷氨酸丙酮酸转氨酶（GPT）、谷氨酸草酰乙酸转氨酶（GOT）、谷氨酰胺合成酶（GS）、游离氨基酸（aa）和谷氨酸合成酶（GOGAT）等活性变...     

Functional expression of a new pharmacological subtype of brain nicotinic acetylcholine receptor

A new type of agonist-binding subunit of rat neuronal nicotinic acetylcholine receptors (nAChRs) was identified. Rat genomic DNA and complementary DNA encoding this subunit (alpha 2) were cloned and analyzed. Complementary DNA expression studies in Xenopus oocytes revealed that the injection of messenger RNAs (mRNAs) for alpha 2 and beta 2 (a neuronal nAChR subunit) led to the generation of a functional nAChR. In contrast to the other known neuronal nAChRs, the receptor produced by the injection of alpha 2 and beta 2 mRNAs was resistant to the alpha-neurotoxin Bgt3.1. In situ hybridization histochemistry showed that alpha 2 mRNA was expressed in a small number of regions, in contrast to the wide distribution of the other known agonist-binding subunits (alpha 3 and alpha 4) mRNAs. These results demonstrate that the alpha 2 subunit differs from other known agonist-binding alpha-subunits of nAChRs in its distribution in the brain and in its pharmacology.     

PirB restricts ocular-dominance plasticity in visual cortex

Experience can alter synaptic connectivity throughout life, but the degree of plasticity present at each age is regulated by mechanisms that remain largely unknown. Here, we demonstrate that Paired-immunoglobulin-like receptor B (PirB), a major histocompatibility complex class I (MHCI) receptor, is expressed in subsets of neurons throughout the brain. Neuronal PirB protein is associated with synapses and forms complexes with the phosphatases Shp-1 and Shp-2. Soluble PirB fusion protein binds to cortical neurons in an MHCI-dependent manner. In mutant mice lacking functional PirB, cortical ocular-dominance plasticity is more robust at all ages. Thus, an MHCI receptor is expressed in central nervous system neurons and functions to limit the extent of experience-dependent plasticity in the visual cortex throughout life. PirB is also expressed in many other regions of the central nervous system, suggesting that it may function broadly to stabilize neural circuits.     

小麦品种麦谷蛋白亚基的遗传变异分析

分析了９６个小麦品种高、低分子量麦谷蛋白亚基组成的及其分布规律,结果表明：强面筋品种与普通品种在高分子量及低分子量麦谷蛋白亚基组成上存在明显差异。强面筋品种含Ｇｌｕ－Ａｌａ（１亚基）、Ｇｌｕ－Ａ１ｂ（２＾＊亚基）、Ｇｌｕ－Ｂ１ｂ（７＋８亚基）、Ｇｌｕ－Ｂｌｉ（１７＋１８亚基）、Ｇｌｕ－Ｄｌｄ（５＋１０亚基）、Ｇｌｕ－Ａ３ｂ和Ｇｌｕ－Ｄ３ｃ的频率较高,而普通品种则含Ｇｌｕ－Ａｌｃ（Ｎ）、Ｇｌｕ－Ｂｌ     

微波对雏鸡的认知功能和NMDA受体亚基基因表达的影响（摘要）

[目的]研究微波对鸡胚生长发育及雏鸡认知功能的影响。[方法]采用磁控管发射2450MHz的微波来模拟微波辐射源辐射种蛋,直到雏鸡孵出后,分别采用一次性被动回避学习和RT-PCR检测微波对雏鸡的认知功能和NMDA受体NR1亚基与NR2亚基表达量的影响。[结果]微波辐射后暴露组的回避率显著低于对照组,特别在辐射强度最高组的回避率极显著低于对照组。同时暴露组有2组雏鸡体重增加,其中一组的孵化时间有所增加。经RT-PCR分析,NR2亚基表达量在第10天而及15天时都上调,NR1亚基表达量只在第15天时才下调。[结论]微波对个体发育具有一定影响,通过使端脑中NMDA受体结构组成和功能发生改变,使自身的调节能力下降,从而对认知功能有一定损害。     

Astrocytes potentiate transmitter release at single hippocampal synapses

Astrocytes play active roles in brain physiology. They respond to neurotransmitters and modulate neuronal excitability and synaptic function. However, the influence of astrocytes on synaptic transmission and plasticity at the single synapse level is unknown. Ca(2+) elevation in astrocytes transiently increased the probability of transmitter release at hippocampal area CA3-CA1 synapses, without affecting the amplitude of synaptic events. This form of short-term plasticity was due to the release of glutamate from astrocytes, a process that depended on Ca(2+) and soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein and that activated metabotropic glutamate receptors (mGluRs). The transient potentiation of transmitter release became persistent when the astrocytic signal was temporally coincident with postsynaptic depolarization. This persistent plasticity was mGluR-mediated but N-methyl-d-aspartate receptor-independent. These results indicate that astrocytes are actively involved in the transfer and storage of synaptic information.     

猪组成型雄烷受体激动剂高通量药物筛选模型的建立

构建猪组成型雄烷受体(pgCAR)激动剂体外高通量筛选模型。利用RT-PCR技术从猪肝脏总RNA中扩增出pgCAR基因序列,将测序后正确的pgCAR片段连接到pcDNA 3.1 vector上构建表达载体pcDNA 3.1-CAR;将合成的5个拷贝NR1(Nuclear receptor sites)插入PGL3-TK-promoter构成报告质粒(NR1)5-TK-luc,与PRL-TK报告质粒构成双荧光报告系统。用PEI转染技术将表达载体与报告质粒共转染HepG 2细胞株中,通过检测荧光素酶基因表达状况评价具有生物活性配体化合物对pgCAR激动活性。阳性药苯妥英钠明显提高荧光素酶的表达,最大上调倍增数可达约2.53倍,并且在一定浓度下阳性药与相对荧光酶的活性表达有较好量效关系。     

浙南小麦核心育种亲本高分子量谷蛋白亚基组成分析

为了解浙南小麦核心育种亲本的品质遗传基础,采用SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)技术对28份核心品种(系)的高分子量谷蛋白亚基(HMW-GS)组成进行分析。结果表明:供试品种(系)在Glu-A1位点具有Null(57.14%)和1(42.86%)2种类型;在Glu-B1位点具有7+8(67.86%)、7+9(25.00%)、17+18(3.57%)和13+16(3.57%)4种类型;在Glu-D1位点具有2+12(96.43%)和5+10(3.57%)2种类型。另外,各供试品种(系)共有5种亚基组合,依次为1/7+8/2+12(39.29%)、Null/7+8/2+12(28.57%)、Null/7+9/2+12(25.00%)、1/17+18/5+10(3.57%)和Null/13+16/2+12(3.57%)。     

中国小麦大面积推广品种及骨干亲本的高分子量谷蛋白亚基组成分析

 对在我国小麦育种中发挥过重要作用的 2 2个骨干亲本、在生产上曾发挥过巨大增产效益的 4 5个品种 ,以及 18个优质面包品种的高分子量谷蛋白亚基组成进行了比较系统的分析。结果表明 ,骨干亲本在 Glu- A1位点有两种等位变异类型“0”和“1”;Glu- B1有“7+8”、“7+9”、“14 +15”、“17+18”、“6 +8”　 5个等位变异类型 ,但主要以“7+8”和“7+9”为主 ;Glu- D1有“2 +12”、“2 +10”、“5 +10”、“4 +10”、“4 +12”、“2 +11” 6个等位变异类型 ,以“2 +12”和“2 +11”为主要类型。在骨干亲本中仅有一个“早洋麦”(Early Prem ium)携带“5 +10”;一个 St2 4 2 2 / 4 6 4携带“14 +15”。此外 ,两个比较重要的材料“玛拉”(Mara)和“阿龙爪”(Alondra)携带“5 +10”。在大面积推广品种中 ,只有“扬麦5号”为“5 +10”携带者 ;“小偃 6号”和“豫麦 7号”携带“14 +15”;“阿夫”、“农大 139”、“郑州 6 83”、“繁 6”等 4个品种携带“17+18”。我国第一批优质小麦品种主要有两个类群 ,即“5 +10”和“14 +15”,前者以“中作 8131- 1”及其变异后代选系为代表 ,后者以“小偃 6号”和其衍生品种为主。此外 ,还查明“17+18”,来源于意大利品种“阿夫” (Funo,或选系 )。上述结果基本反映了 5 0     

Driving AMPA receptors into synapses by LTP and CaMKII: requirement for GluR1 and PDZ domain interaction

To elucidate mechanisms that control and execute activity-dependent synaptic plasticity, alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptors (AMPA-Rs) with an electrophysiological tag were expressed in rat hippocampal neurons. Long-term potentiation (LTP) or increased activity of the calcium/calmodulin-dependent protein kinase II (CaMKII) induced delivery of tagged AMPA-Rs into synapses. This effect was not diminished by mutating the CaMKII phosphorylation site on the GluR1 AMPA-R subunit, but was blocked by mutating a predicted PDZ domain interaction site. These results show that LTP and CaMKII activity drive AMPA-Rs to synapses by a mechanism that requires the association between GluR1 and a PDZ domain protein.     

Direct evidence that endogenous GM1 ganglioside can mediate thymocyte proliferation

The B subunit of cholera toxin, which is multivalent and binds exclusively to a specific ganglioside, GM1, was mitogenic for rat thymocytes. When exposed to the B subunit, the cells proliferated, as measured by 3H-labeled thymidine incorporation. Mitogenesis depended on the direct interaction of the B subunit with GM1 on the surface of the cells. This demonstrates that endogenous plasma membrane gangliosides can mediate proliferation in lymphocytes.     

SV2 is the protein receptor for botulinum neurotoxin A

How the widely used botulinum neurotoxin A (BoNT/A) recognizes and enters neurons is poorly understood. We found that BoNT/A enters neurons by binding to the synaptic vesicle protein SV2 (isoforms A, B, and C). Fragments of SV2 that harbor the toxin interaction domain inhibited BoNT/A from binding to neurons. BoNT/A binding to SV2A and SV2B knockout hippocampal neurons was abolished and was restored by expressing SV2A, SV2B, or SV2C. Reduction of SV2 expression in PC12 and Neuro-2a cells also inhibited entry of BoNT/A, which could be restored by expressing SV2 isoforms. Finally, mice that lacked an SV2 isoform (SV2B) displayed reduced sensitivity to BoNT/A. Thus, SV2 acts as the protein receptor for BoNT/A.     

钼对甘蔗氮代谢的影响

本试验选用巴西固氮甘蔗品种B 1和B 8为供试材料,在温室条件下桶栽砂培甘蔗施以含不同钼水平的营养液(0m g/L、0.01m g/L、0.02m g/L、0.04m g/L),在不同时期测定甘蔗叶片中硝酸还原酶、G S酶活性及硝态氮、氨态氮含量。试验结果表明,在大棚含氮条件下,三个钼处理都提高了两个甘蔗品种的硝酸还原酶活性而降低了硝态氮含量。0.01m g/L钼处理在拔节期后提高了B 1叶片的G S酶活性,而在幼苗期提高了B 8叶片的G S酶活性,降低了氨态氮含量,促进了氮代谢。在大棚无氮条件下,三个钼处理在拔节期后都提高了两个甘蔗品种的NR酶活性,降低硝态氮含量;提高了B 1的G S酶活性,降低氨态氮含量;在分蘖期也提高了B 8叶片的G S酶活性而降低了其氨态氮含量。因而在大棚里,无论是含氮或无氮条件下,在不同时期不同钼处理对甘蔗氮代谢和氨同化过程的作用也不同,但是相对以较低水平钼处理较能提高两个甘蔗品种的硝酸还原酶活性而降低硝态氮含量;提高了B 1与B 8的G S酶活性,降低氨态氮含量,促进了甘蔗的氮代谢和氨同化。     

施氮量和种植密度对优质晚稻稻米品质的调控效应及其机制

以‘玉针香’为材料,设计4个施氮量处理,分别为N1(不施氮)、N2(尿素180 kg/hm2)、N3(尿素126 kg/hm2)、N4(180 kg/hm2尿素+生物炭200 t/hm2),2个种植密度处理,分别为D1(18 cm×25 cm)、D2(14 cm×25 cm),于2020—2021年开展大田试验,研究施氮量、种植密度及其互作对稻米品质与叶片氮代谢特征的影响。结果表明：种植密度对稻米出米率、直链淀粉含量、胶稠度、蛋白质组分均无显著影响,D2处理各指标均略高于D1处理;施氮提高了出米率与各蛋白质组分含量,降低了垩白粒率和垩白度,以N4处理的效果最好;施氮量与种植密度互作对稻米品质影响显著,N4D2处理的稻米加工品质与外观品质最优,胶稠度最长,蛋白质组分含量最高;种植密度对叶片含氮量、硝态氮含量以及硝酸还原酶(NR)、亚硝酸还原酶(NiR)活性均无显著影响;D2处理的叶片含氮量以及NR、NiR活性较高;施氮提高了叶片全氮含量、硝态氮含量以及NR、NiR活性,以N4处理的最高;相关性分析结果表明,叶片全氮含量、NR活性、NiR活性与整精米率、直链淀粉含量、胶稠度、蛋白组分含量呈显著或极显著正相关,与垩白粒率和垩白度呈显著或极显著负相关;增施生物炭可显著提高整精米率,显著降低垩白度和垩白粒率。本试验条件下,N4D2处理对优质稻稻米品质的调控效应最好,其调控机制在于提高了叶片全氮含量和氮代谢酶活性。