Kinesin superfamily motor protein KIF17 and mLin-10 in NMDA receptor-containing vesicle transport

Experiments with vesicles containing N-methyl-D-aspartate (NMDA) receptor 2B (NR2B subunit) show that they are transported along microtubules by KIF17, a neuron-specific molecular motor in neuronal dendrites. Selective transport is accomplished by direct interaction of the KIF17 tail with a PDZ domain of mLin-10 (Mint1/X11), which is a constituent of a large protein complex including mLin-2 (CASK), mLin-7 (MALS/Velis), and the NR2B subunit. This interaction, specific for a neurotransmitter receptor critically important for plasticity in the postsynaptic terminal, may be a regulatory point for synaptic plasticity and neuronal morphogenesis.     

Effects of cold stress on hypothalamic corticotrophin-releasing and thyrotropin-releasing hormone messenger RNA levels in chickens

There is increasing evidence that stress can activate the hypothalamic-pituitary-adrenal-axis and hypothalamic-pituitary- thyroid-axis,and further affect the synthesis and secretion of corticotrophin-releasing hormone(CRH)and thyrotropin-releasing hormone(TRH).To evaluate the effect of cold stress on the hypothalamic CRH and TRH messenger RNA(mRNA)levels in Yisha chickens,male Yisha chickens were subjected to acute(1,6,12 h)and chronic(5,10,20 d)cold stress(12±1)℃.Hypothalami were collected for assessment of mRNA levels by semi-quantitative RT-PCR.Acute stress resulted in a significant decrease of CRH mRNA levels at 6 and 12 h,and a significant increase of TRH mRNA levels at every stress time point.Chronic cold stress resulted in a significant increase of CRH mRNA levels and a significant decrease of TRH mRNA levels compared with the control group at every stress time point.The results suggest that the two genes differently respond to cold stress at the mRNA levels.And the different degrees of cold stress will produce different effects on the identical gene.     

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...     

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.     

Ca2+参与硝普钠(SNP)对干旱胁迫下小麦幼苗叶片NO水平的调控

研究干旱胁迫下外源一氧化氮(NO)供体硝普钠(SNP)处理对小麦幼苗叶片NO水平的影响及其与Ca2+的关系.在15%PEG-6000胁迫下测定NO含量和NO合成酶活性,结果表明,干旱胁迫下小麦幼苗叶片一氧化氮合酶(NOS)活性显著增加,且钙依赖型cNoS(CaM的组成型NOS)快速调控NO产生,但是随着胁迫时间的延长,不依赖钙iNOS(CaM的诱导型NOS)的活性在NOS活性比例缓慢增加,而硝酸还原酶(NR)产生NO的能力只占总NR提取物活性的很小一部分;0.1 mmol/L SNP处理可显著提高干旱胁迫下小麦幼苗叶片NOS和NR活性,诱导NO水平提高,显著缓解膜脂过氧化;用质膜Ca2+通道抑制剂LaCl3与SNP共处理,显著减弱或抵消SNP促进NO合成作用.SNP显著提高干旱胁迫下小麦幼苗叶片NO合成酶活性和NO含量,有效缓解膜的氧化损伤,而Ca2+参与SNP对干旱胁迫下小麦幼苗叶片NO水平的调控.     

Ethanol augments GABAergic transmission in the central amygdala via CRF1 receptors

The central amygdala (CeA) plays a role in the relationship among stress, corticotropin-releasing factor (CRF), and alcohol abuse. In whole-cell recordings, both CRF and ethanol enhanced gamma-aminobutyric acid-mediated (GABAergic) neurotransmission in CeA neurons from wild-type and CRF2 receptor knockout mice, but not CRF1 receptor knockout mice. CRF1 (but not CRF2) receptor antagonists blocked both CRF and ethanol effects in wild-type mice. These data indicate that CRF1 receptors mediate ethanol enhancement of GABAergic synaptic transmission in the CeA, and they suggest a cellular mechanism underlying involvement of CRF in ethanol's behavioral and motivational effects.     

球孢白僵菌硝酸还原酶基因同源重组载体的构建

根据已报道球孢白僵菌(Beauveria bassiana)硝酸还原酶基因(nitrate reductase,NR)序列设计引物,从球孢白僵菌D1-5菌株基因组DNA中扩增并克隆该基因上下游片段NR1和NR2,成功构建了球孢白僵菌NR基因同源重组敲除载体,以期敲除球孢白僵菌的NR基因,建立白僵菌同源重组基因敲除体系。     

Neurokinin 1 receptor antagonism as a possible therapy for alcoholism

Alcohol dependence is a major public health challenge in need of new treatments. As alcoholism evolves, stress systems in the brain play an increasing role in motivating continued alcohol use and relapse. We investigated the role of the neurokinin 1 receptor (NK1R), a mediator of behavioral stress responses, in alcohol dependence and treatment. In preclinical studies, mice genetically deficient in NK1R showed a marked decrease in voluntary alcohol consumption and had an increased sensitivity to the sedative effects of alcohol. In a randomized controlled experimental study, we treated recently detoxified alcoholic inpatients with an NK1R antagonist (LY686017; n = 25) or placebo (n = 25). LY686017 suppressed spontaneous alcohol cravings, improved overall well-being, blunted cravings induced by a challenge procedure, and attenuated concomitant cortisol responses. Brain functional magnetic resonance imaging responses to affective stimuli likewise suggested beneficial LY686017 effects. Thus, as assessed by these surrogate markers of efficacy, NK1R antagonism warrants further investigation as a treatment in alcoholism.     

Autocrine or paracrine inflammatory actions of corticotropin-releasing hormone in vivo

Corticotropin-releasing hormone (CRH) functions as a regulator of the hypothalamic-pituitary-adrenal axis and coordinator of the stress response. CRH receptors exist in peripheral sites of the immune system, and CRH promotes several immune functions in vitro. The effect of systemic immunoneutralization of CRH was tested in an experimental model of chemically induced aseptic inflammation in rats. Intraperitoneal administration of rabbit antiserum to CRH caused suppression of both inflammatory exudate volume and cell concentration by approximately 50 to 60 percent. CRH was detected in the inflamed area but not in the systemic circulation. Immunoreactive CRH is therefore produced in peripheral inflammatory sites where, in contrast to its systemic indirect immunosuppressive effects, it acts as an autocrine or paracrine inflammatory cytokine.     

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。     

表油菜素内酯对水分胁迫下菘蓝相关指标的影响

目的:研究水分胁迫下表油菜素内酯(EBR)对菘蓝相关酶活性和生理指标的影响,为提高菘蓝的抗涝能力提供方法。方法:采用盆栽试验,测定水分胁迫下,不同浓度EBR(0.01、0.050、.10、0.501、.00 mg/L)处理下的菘蓝的抗氧化保护酶(SOD、POD、CAT、APX)、乙醇脱氢酶(ADH)、硝酸还原酶(NR)活性和脯氨酸(Pro)、可溶性蛋白、丙二醛(MDA)的含量。结果:当EBR浓度为0.100、.50 mg/L时,菘蓝的黄叶数最少,随着EBR浓度的增加,抗氧化酶、ADH、NR活性先增后降,Pro、可溶性蛋白含量也呈相同变化趋势,MDA含量变化则与之相反。结论:适宜浓度的EBR能显著提高菘蓝的抗氧化能力,减少水分胁迫造成的伤害。     

CRH、UⅠ和UⅡ基因在斑马鱼发育早期的表达谱

促肾上腺皮质激素释放激素(Corticotropin-releasing hormone,CRH)、尾加压素Ⅰ(UrotensinⅠ,UⅠ)和尾加压素Ⅱ(UrotensinⅡ,UⅡ)广泛存在于脊椎动物中。在鱼类中,它们在渗透压调节、繁殖、营养代谢以及应激反应中发挥着重要作用。本研究中,通过收集0、1.5、3.7、5.3、8、10、16、24、48、72、96、120和144 hpf(hours post-fertilization)的斑马鱼早期发育样品,运用荧光定量PCR技术进行检测发现CRH、UⅠ、UⅡα和UⅡβmRNA从0 hpf开始存在表达,而且CRH、UⅠ、UⅡα和UⅡβ的表达量随发育而逐渐增加,并分别在120 hpf或144 hpf达到最高水平。相关性分析揭示在斑马鱼早期发育过程中,CRH、UⅠ、UⅡα和UⅡβmRNA的表达水平两两之间呈现极显著正相关,这表明CRH、UⅠ、UⅡα和UⅡβ在斑马鱼早期发育过程中关系紧密并且相互合作发挥作用。     

Effect of Salt Stress on Nitrogen Assimilation of Functional Leaves and Root System of Rice in Cold Region

The aims were to investigate the effect of salt stress on key enzyme activity of nitrogen metabolism and the concentration of nitrate nitrogen and ammonium nitrogen response to salt stress.Two rice cultivars,Mudanjiang 30 (sensitive cultivar) and Longdao 5 (salt-tolerant cultivar),were treated with different salt concentrations (CK 0%,S1 0.075%,S2 0.15%,S3 0.225% and S4 0.3%).The results showed that the activities of nitrate reductase (NR),glutamine synthase (GS),glutamate synthase (GOGAT) and glutamate dehydrogenase (GDH) in the functional leaves and roots of rice in cold region presented a single peak curve change and the peak occurred in the heading stage;compared with those of the CK,the activities of NR,GS and GOGAT of rice in cold region decreased,but the activity of GDH increased in the heading stage under salt stress.The variation for key enzyme activity of nitrogen metabolism was the highest under S4 treatment.The activities of NR,GS and GOGAT in the functional leaves significantly decreased compared with those in roots;the concentrations of nitrate nitrogen and ammonium nitrogen in the functional leaves and roots of rice in cold region presented a single peak curve change and the peak occurred in the heading stage;compared with that of the CK,the concentration of nitrate nitrogen decreased in leaves and roots,the concentration of ammonium nitrogen decreased and the concentration of ammonium nitrogen in roots increased under salt stress.The variations for the activities of NR,GS and GOGAT in the functional leaves and roots of Longdao 5 were less than those of Mudanjiang 30 under the same concentration of salt stress.     

Shade stress decreases stem strength of soybean through restraining lignin biosynthesis

Lodging is the most important constraint for soybean growth at seedling stage in maize-soybean relay strip intercropping system. In the field experiments, three soybean cultivars Nandou 032-4 (shade susceptible cultivar; B1), Jiuyuehuang (moderately shade tolerant cultivar; B2), and Nandou 12 (shade tolerant cultivar; B3) were used to evaluate the relationship between stem stress and lignin metabolism in the stem of soybean. Results showed that the intercropped soybean was in variable light condition throughout the day time and co-growth stage with maize. The xylem area and cross section ratio played a main role to form the stem stress. The B3 both in intercropping and monocropping expressed a high stem stress with higher xylem area, lignin content, and activity of enzymes (phenylalanine ammonia-lyase (PAL), 4-coumarate: CoA ligase (4CL), cinnamyl alcohol dehydrogenase (CAD), and peroxidase (POD)) than those of B1 and B2. Among the soybean cultivars and planting pattern, lignin content was positively correlated with stem stress. However, a negative correlation was found between lignin content and actual rate of lodging. In conclusion, the shade tolerant soybean cultivar had larger xylem area, higher lignin content and activities of CAD, 4CL, PAL, and POD than other soybean cultivars in intercropping. The lodging in maize-soybean intercropping can be minimized by planting shade tolerant and lodging resistant cultivar of soybean. The lignin content in stem could be a useful indicator for the evaluation of lodging resistance of soybean in intercropping and activities of enzymes were the key factors that influence the lignin biosynthesis.     

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。     

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.     

运输应激对刀鲚生理生化指标和HPI轴基因表达影响及甘草甜素的作用

以刀鲚(Coilia nasus)为研究对象,运用荧光定量和相关生化测定方法,研究运输应激对下丘脑-垂体-肾间组织轴(HPI)基因表达和肝脏氧化指标的影响;同时采用浸泡的方式研究甘草甜素(glycyrrhizin,GL)对以上指标的调控作用。结果显示:运输后,脑中促肾上腺皮质激素释放激素(CRH)基因、阿黑皮素原(POMC)基因和硬骨鱼紧张肽(UI)基因的表达水平显著降低;头肾中皮质醇受体(GR)基因的表达水平运输2 h明显下降,4 h时表达水平最低,8 h显著升高。运输应激可以明显降低肝脏过氧化氢酶(CAT)的活性,显著提高肝脏丙二醛(MDA)的水平,而对谷胱甘肽过氧化物酶(GSH-PX)和脂质过氧化物(LPO)无显著影响。添加甘草甜素6 h后头肾GR基因的表达水平降低,脑中CRH和POMC基因的表达水平显著升高,但是脑中UI基因的表达水平无明显变化。此外,添加甘草甜素对肝脏CAT、MDA、GSH-PX和LPO无显著影响。以上结果表明,运输应激调控HPI轴的基因表达发生了不同程度的变化,而且降低了机体的抗氧化能力,使刀鲚处于氧化应激状态中。GL也可以影响HPI轴的基因表达,但不能有效缓解刀鲚的运输应激引发的氧化应激。     

黄瓜幼苗耐低温指标研究初报

通过研究低温对黄瓜幼苗的影响，发现在低温条件下，黄瓜幼苗叶片叶绿素含量明显下降，其相对减少量与黄瓜耐低温能力呈负相关；大部分品种（系）可溶性蛋白质含量增加，低温下可溶性蛋白质含量及相对增加量与黄瓜耐低温能力表现为正相关。在低温条件下保持较高的叶绿素含量和ＮＲ活性是黄瓜植株保持较大生长量的基础。经相关分析，ＮＲ活性及其相对减少量与黄瓜耐低温能力呈显著相关，是鉴定黄瓜耐温能力的较好指标。并对植物耐低温能力的复杂性做了分析。     

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。