乙酰胆碱对黄化玉米幼苗中胚轴通透性与物质运转的凋控效应

对乙酰胆碱处理前后的黄化玉米（Zea mays L.）幼苗中胚轴切段节和节间的通透性和物质运转进行了比较。用电阻法测定通透性；通过加在胚芽鞘切口的经共质体转运的特异荧光物质羧基荧光素（carboxyfluorescein,CF)的分布检察物质运转。节间由柱状的、排列方向多少和长轴方向平行的细胞构成；而构成节的细胞短、排列不规则。结果表明跨节比电阻比节间的高，通透性低的短的节细胞比柱状节间细胞累积更多的CF。0.1mmol/L乙酰胆碱（ACh）处理增加了节和节间的通透性，促进了CF的运转。物质在节和节间运转的不同，以及ACh对此的调控效应很大程度上可归结于细胞通透性的差异。     

乙酰胆碱通过作用于α7烟碱型乙酰胆碱受体抑制大鼠小胶质细胞炎性反应

小胶质细胞是中枢神经系统中主要的免疫细胞。本研究旨在探讨乙酰胆碱(acetylcholine,ACh)抑制小胶质细胞炎症反应的具体机制。原代培养Sprague-Dawley(SD)大鼠小胶质细胞,脂多糖(lipopolysaccharide,LPS)诱导建立炎症反应模型,ACh处理24 h后,用Western blot检测多种炎性因子、胰岛素样生长因子1(insulin-like growth factor 1,IGF-1)和α7烟碱型乙酰胆碱受体(α7 nicotinic acetylcholine receptor,α7nAChR)的蛋白表达,用ELISA检测多种炎性因子和IGF-1的释放情况,用慢病毒转染沉默α7nAChR后观察ACh作用的变化。结果显示,LPS可促进小胶质细胞的激活,上调诱生型一氧化氮合酶(inducible nitric oxide synthase,iNOS)蛋白表达,增加白介素1β(interleukin-1β,IL-1β)和肿瘤坏死因子α(tumor necrosis factorα,TNF-α)的表达和释放,减少神经营养因子IGF-1的表达和释放,ACh能逆转LPS的这些作用;LPS可下调小胶质细胞的α7nAChR蛋白表达,ACh逆转该作用;α7n AChR-shRNA慢病毒转染小胶质细胞后,ACh对抗LPS的作用消失。以上结果提示,ACh对抗LPS诱导的小胶质细胞炎症反应的保护作用是通过α7nAChR实现的,这为今后神经炎症疾病的治疗提供了新的治疗靶点。     

豇豆螺旋茎段维管组织观察及其成因分析

豇豆(Vigna unguiculata Linn.)的茎缠绕支持物形成右手螺旋,其茎段经水浸泡去表皮后可见维管纤维沿茎轴方向呈左手螺旋状排列.用徒手切片法观察螺旋型和直线型茎段各节间中部的横切面,发现二者木质部导管分子均由内至外呈对称的扇形排列;螺旋型茎段各节间扇形的角度均显著小于直线型茎段对应节间的扇形角度(p＜0.05);除第1节间外,螺旋型茎段各节间扇形纵横比均显著大于直线型茎段对应节间的扇形纵横比(p＜0.05).分析表明,茎的两种螺旋之间存在扭力平衡;螺旋型与直线型茎段横切面导管分子扇形排列的差异可能是茎段扭转过程中受挤压所致.此外,茎段扭转并不是细胞横向不对称扩张的结果,而可能是由各侧细胞不均衡的纵向生长所造成.     

胚胎型与成年型肌肉乙酰胆碱受体分子有区别

电鱼N 型乙酰胆碱(ACh)受体是由五条多肽链:α_2βγδ组成。这四种亚基构成一个离子通道。现已弄清这四个亚基的一级结构。当两个ACh 分子与α亚基的细胞外受点结合时,激活离子通道,完成一个快速的化学传递。哺乳动物其ACh 受体的生理特性随胚胎发育的进行而不同。在运动神经开始对肌肉支配之     

AChE在水稻抗性诱导的褐飞虱凋亡中肠细胞中的定位及表达

【目的】褐飞虱Nilaparvata lugens是水稻的重要害虫之一。本研究旨在了解水稻品种抗性诱导的褐飞虱中肠细胞凋亡与细胞中乙酰胆碱酯酶(acetylcholinesterase,ACh E)的关系。【方法】从4龄Ⅰ型褐飞虱若虫中肠组织分离原代细胞,用不同抗性的水稻幼苗汁液处理第一代继代细胞,利用TUNEL染色法检测细胞的凋亡情况,再分别利用免疫组织化学和荧光定量PCR技术对ACh E进行亚细胞定位和检测其表达水平的变化。【结果】在对照组(未处理)细胞中,检测不到代表凋亡细胞核的绿色荧光,而免疫组化后阳性反应颜色很浅,表明细胞中存在ACh E的本底水平表达。感虫水稻品种TN1幼苗汁液处理细胞中,细胞的凋亡率为8%,抗性水稻B5幼苗汁液处理的细胞中有65%的细胞发生凋亡;而抗性水稻TKM-6幼苗汁液处理的细胞中则有85%的细胞发生凋亡。免疫组化的检测结果表明,所有凋亡的细胞中都存在ACh E的积累,且主要分布在细胞质中;ACh E在凋亡后期并不迁入凋亡小体中,而是集中在细胞核附近。荧光定量PCR检测表明,TKM-6,B5和TN1幼苗汁液处理的细胞中ACh E的表达量分别为对照细胞的29.9,18.4和8倍,该结果与细胞水平上免疫组化的检测结果一致。【结论】本研究结果证实了水稻品种抗性与其诱导的中肠细胞凋亡率呈正相关,且凋亡细胞的细胞质中存在ACh E的积累。这些发现为揭示ACh E在褐飞虱与水稻的互作中的功能、促进抗性水稻新品种的选育及开发新的褐飞虱防治措施提供了一定的参考信息。     

指数施肥对白桦裸根苗生长动态、生物量分配及光合作用的影响

设定常规施肥(CF)、指数施肥(EF)和1.5倍指数施肥(EF1.5)3种施肥处理研究不同氮素施入方法和剂量对白桦1年生裸根苗生长(苗高、地径、生物量)动态、生物量分配、相对生长速率(RGR)和光合作用的影响。研究结果表明:(1)3种处理苗木生长前期苗高、地径和生物量均无显著差异,生长后期EF与EF1.5显著高于CF(P<0.05),EF与EF1.5间差异不显著(P>0.05);(2)生长结束时,EF、EF1.5处理苗高分别比CF高18.2%和25%(P>0.05),地径分别比CF高11.2%和5.8%(P<0.05),单株生物量分别比CF高41.5%和25.4%(P<0.05);(3)EF与EF1.5处理苗木的茎、叶、根生物量均显著高于CF,各处理间苗木的根冠比差异不显著(P>0.05);(4)EF1.5处理和EF处理苗木苗高、地径及生物量相对生长速率(RGR)均显著高于CF处理(P<0.05),而EF1.5处理和EF处理间仅生物量相对生长速率达到显著水平,且EF>EF1.5(P<0.05);(5)苗木生长高峰期,EF处理提高了苗木叶片的光合能力。EF处理苗木叶面积高于CF处理16.8%(P<0.05),叶绿素含量及光合速率较CF处理分别提高了4.5%(P>0.05)、5.0%(P>0.05)。EF、EF1.5处理常规施肥处理促进了苗木生长和生物量积累,二者促进效果类似,认为EF为最佳的施肥处理。     

猴和猫初级视皮层细胞方向选择性及其功能组织的比较研究

虽然许多视皮层区内的方向选择性细胞以柱状方式排列组构, 但猴初级视皮层(V1)区方向选择性功能组构仍然不甚明了. 定量地比较研究了猴和猫初级视皮层方向选择性细胞的比例、选择性强度和功能组织, 结果表明, 猴的方向选择性细胞的比例比猫少, 选择性强度和方向功能组织的程度均显著地比猫弱, 提示这些种属差别可能源于其神经通路的不同.     

乙酰胆碱对自然杀伤细胞活性的影响

目的:观察乙酰胆碱(ACh)对自然杀伤(NK)细胞活性的影响,并初步探讨其作用的受体机制.方法:根据不同的实验目的,选择ACh、胆碱能受体激动剂和拮抗剂分别作用于NK细胞,以乳酸脱氢酶(lactate dehydrogenase,LDH)自然释放法检测不同实验条件下NK细胞杀伤肿瘤靶细胞(Yac)的活性.结果:ACh、M受体激动剂毛果芸香碱和N受体激动剂烟碱在10-10～10-6mol/L浓度范围内都能显著抑制NK细胞杀伤肿瘤细胞的活性.M受体拮抗剂阿托品(10-8和10-7mol/L)能完全阻断同浓度ACh抑制NK细胞活性的作用;但N受体拮抗剂筒箭毒碱(10-8和10-7mol/L)不能阻断同浓度ACh抑制NK细胞活性的作用.结论:ACh可抑制NK细胞对肿瘤细胞的杀伤作用,此作用主要由淋巴细胞上的M受体和N1受体介导.     

全反式维甲酸对乙酰胆碱受体特异性淋巴细胞的免疫调控作用

目的:观察全反式维甲酸(ATRA)对乙酰胆碱受体(AChR)特异性淋巴细胞的体外调控作用,探讨其治疗重症肌无力(MG)的可能机制。方法:建立完全弗氏佐剂(CFA)对照组及实验性自身免疫性重症肌无力(EAMG)组大鼠,并获取淋巴结单个细胞悬液,以ACh R97-116多肽片段以及不同浓度的ATRA体外培养72 h,采用流式细胞仪法、CCK-8法、ELISA法分别检测活细胞比例、细胞凋亡和周期的改变以及Th亚群的格局和B细胞抗体分泌能力的变化。结果:ATRA显著降低活细胞比例(P0.001);不同浓度的ATRA均促进了特异性细胞群的凋亡(P0.001),且呈剂量依赖性,而ATRA未改变AChR特异性淋巴细胞的生长周期;ATRA处理后,CFA和EAMG组的淋巴细胞增殖均受到明显抑制,且ATRA对ACh R特异性的淋巴细胞的抑制明显(EAMG组,P0.01)于CFA组(P0.05);ATRA干预后,ACh R特异性CD4+T淋巴细胞的比例下降(P0.01),且ATRA促进了Th2、Treg细胞亚群百分比(P_(IL-4)0.001,P_(Foxp3)0.001),而抑制了促炎性的Th17、Th1细胞亚群百分比(P_(IL-17)0.05,P_(IFN-γ)0.001);ATRA能够降低ACh R特异性B细胞的抗体分泌能力(P0.01)。结论:ATRA不仅能抑制ACh R特异性T细胞功能,同时也能抑制ACh R特异性B细胞功能,其在MG的临床治疗中可能起治疗作用。     

大鼠肠道内NOS与AChE、VIP阳性神经元的分布关系研究

应用一氧化氮合酶 (NOS)、乙酰胆碱酯酶 (ACh E)组织化学及血管活性肠肽 (VIP)免疫组织化学方法 ,光镜下比较观察大鼠肠道内 NOS、ACh E、VIP阳性神经元的形态学特征。结果显示 ,肠肌间丛 NOS阳性神经元胞体大小不等 ,形态不一 ,NOS、ACh E和 VIP阳性神经元的分布密度为 ACh E>NOS>VIP,在不同的肠段和层次分布密度有差异 ,NOS与 ACh E存在共染。在肌间丛和粘膜下丛 ,少数 VIP与 NOS共染。在粘膜下丛 ,三种阳性神经元的分布密度为 ACh E>VIP>NOS。在肌间丛和粘膜下丛 ,可见 VIP阳性末梢环抱 NOS阳性神经元胞体 ,两者呈终扣样接触。上述结果提示 NOS阳性神经元与 ACh E、 VIP阳性神经元有密切的形态学联系。在消化道功能调节上 ,它们可能起协调作用。     

Intercellular communication in Chara: factors affecting transnodal electrical resistance and solute fluxes

The permeability of plasmodesmata in the nodal complex of branch cells of Chara corallina was examined by measuring both the transnodal electrical resistance and transnodal fluxes of ³⁶CI and ¹⁴C-buty-rate. Under normal circumstances, the resistance across the node was low, but increased rapidly in response to metabolic inhibition, pressure gradients across the node or excision of one of the cells. For each of these treatments, there was a substantial reduction in solute transport between the cells. Acidification of the cytoplasm by weak acids or alkalinization by amines did not affect either the electrical resistance or the flux of solutes through the node between whorl cells. The transnodal resistance was significantly higher in older cell pairs, but was unaffected by large transnodal voltage differences or by the passage of action potentials. There was no evidence that short-term increases in cytoplasmic calcium have any effect on plasmodesmatal permeability.     

Transcellular translocation of Campylobacter jejuni across human polarised epithelial monolayers

The mechanisms whereby Campylobacter jejuni translocates across the host intestinal epithelium are not yet understood and the transepithelial route remains undefined. During C. jejuni translocation, the transmonolayer electrical resistance (TER) across polarised monolayers of Caco-2 cells is not affected and the penetration of [(14)C]inulin across the monolayers does not increase. Over 24 h, however, bacteria damage the monolayer integrity, causing a decrease in the TER. These results support C. jejuni translocation through the cytoplasm of invaded cells (transcellular) rather than via intercellular spaces (paracellular).     

Distribution of Intercellular Material in the Apical Part of Pea Seedlings

The sections from the upper part of the third internode, counted from the seed, in etiolated pea seedlings are studied for the distribution of two types of Intercellular spaces: the transparent and the dark. The transparent spaces represent the result of the water logging of passages originally water-lined, while the dark spaces are lined with a lipid-containing substance which may be impregnated with melted paraffin and which forms a ramifying network within the tissue. The intercellular material of the dark spaces has the appearance of a tube, since it concentrically lines the intercellular space, leaving a lumen for air or gas. It is a plastic, isotropic subslance which may be cut transversely and identified in successive sections of the inlernode as belonging to the same continuous material from the intercellular space. The dark and the transparent spaces have a distinct pattern of distribution in the growing internode, and the relative quantity of dark spaces present at different levels of the internode may be measured. The dark spaces predominate in those parts of the stem with a greater potential for growth, while the transparent spaces are located in the regions where growth has ceased or subsided. Since the paraffin is infiltrated instantaneously throughout the network of dark intercellular spaces, it is possible that these may represent a channel for the translocation of substances.     

Etiolation of stock plants affects adventitious root formation and hormone content of pea stem cuttings

Stock pea plants (Pisum sativum L.) were etiolated fully or partially at the third internode that acted as the cutting base. The etiolation started the fifth day after sowing and lasted till cutting preparation. Cuttings derived from partially etiolated plants rooted more than non-etiolated ones while fully etiolated ones rooted more only after treatment with 1% sucrose solution for 4 days. Endogenous IAA in the base of etiolated cuttings was higher during the first 24 h after cutting preparation than in the control. Z/ZR did not show significant differences while iAde/iAdo was higher in the control. Ethylene was increased 24 h after cutting preparation and the increase was greater from partially etiolated cuttings. The results showed that besides IAA and cytokinins, which played a role in the rooting of cuttings, sucrose influenced rooting in the case of fully etiolated stock plants.     

Effects of human neutrophil chemotaxis across human endothelial cell monolayers on the permeability of these monolayers to ions and macromolecules

We have developed a method for studying the permeability properties of human endothelia in vitro. Human umbilical vein endothelial cells (HUVEC) were cultured on a substrate of human amnion. Confluent monolayers of these cells demonstrated 6-12 delta.cm2 of electrical resistance (a measure of their permeability to ions) and restricted the transendothelial passage of albumin from their apical to their basal surface. To determine whether leukocyte emigration alters endothelial permeability in this model, we examined the effects of migrating human polymorphonuclear leukocytes (PMN) on these two parameters. Few PMN migrated across the HUVEC monolayers in the absence of chemoattractants. In response to chemoattractants, PMN migration through HUVEC monolayers was virtually complete within 10 minutes and occurred at random locations throughout the monolayer. PMN migrated across the monolayer via the paracellular pathway. Although one PMN migrated across the monolayer for each HUVEC, PMN migration induced no change in electrical resistance or albumin permeability of these monolayers. At this PMN:HUVEC ratio, these permeability findings were correlated morphologically to measurements that HUVEC paracellular pathway size increases by less than 0.22% with PMN migration. This increase is insufficient to effect a measurable change in the electrical resistance of the endothelial cell monolayer. These findings demonstrate that increased permeability of cultured endothelial cell monolayers is not a necessary consequence of PMN emigration.     

Neutrophil migration across a cultured epithelial monolayer elicits a biphasic resistance response representing sequential effects on transcellular and paracellular pathways

Migration of polymorphonuclear leukocytes across epithelia is a hallmark of many inflammatory disease states. Neutrophils traverse epithelia by migrating through the paracellular space and crossing intercellular tight junctions. We have previously shown (Nash, S., J. Stafford, and J.L. Madara. 1987. J. Clin. Invest. 80:1104-1113), that leukocyte migration across T84 monolayers, a model human intestinal epithelium, results in enhanced tight junction permeability--an effect quantitated by the use of a simple, standard electrical assay of transepithelial resistance. Here we show that detailed time course studies of the transmigration-elicited decline in resistance has two components, one of which is unrelated to junctional permeability. The initial decrease in resistance, maximal 5-13 min after initiation of transmigration, occurs despite inhibition of transmigration by an antibody to the common beta subunit of neutrophil beta 2 integrins, and is paralleled by an increase in transepithelial short-circuit current. Chloride ion substitution and inhibitor studies indicate that the early-phase resistance decline is not attributable to an increase in tight junction permeability but is due to decreased resistance across epithelial cells resulting from chloride secretion. Since T84 cells are accepted models for studies of the regulation of Cl- and water secretion, our results suggest that neutrophil transmigration across mucosal surfaces (for example, respiratory and intestinal tracts) may initially activate flushing of the surface by salt and water. Equally important, these studies, by providing a concrete example of sequential transcellular and paracellular effects on transepithelial resistance, highlight the fact that this widely used assay cannot simply be viewed as a direct functional probe of tight junction permeability.     

Electrotonic Coupling between Adjacent Internodal Cells of Chara braunii: Transmission of Action Potentials beyond the Node

Many nodal cells are interposed between two internodal cellsof Chara braunii. When an action potential conducted in an internodereached the node, no electrical activation in the nodal cellscould be found, although an area of the membrane bordering thenodal cells in this internode was partially activated (end-membraneaction potential). When the action potential approached thenode along the stimulated internode, an electrotonic potentialchange (depolarization) was produced in the other internode.This depolarization was greatly depressed by the end-membraneaction potential of the stimulated internode, so that hardlyany transmission took place. The ratio of the potential changein the surface membrane of the adjoining ("postsynaptic") internode(cell b) to that of the stimulated one (cell a), the couplingratio, e_b/e_a, can be estimated from a simple equivalent circuitof the nodal region composed of two surface-membrane resistances(R_a, R_b) and intercellular resistance (R_n). If R_n remains thesame, a higher ratio should be produced with a larger R_b, butthe ratio does not depend on any change in R_a, which could beproved experimentally. Transmission of the action potentialbeyond the node was frequent when the coupling ratio was increasedand when the threshold that elicits the action potential waslowered by immersing the node in a K or Na salt solution. ¹ Present address: Department of Physiology, Tohoku UniversitySchool of Dentistry, Sendai 980, Japan. (Received December 1, 1980; Accepted January 23, 1981)     

Functional significance of differential eNOS translocation

Nitric oxide (NO) regulates flow and permeability. ACh and platelet-activating factor (PAF) lead to endothelial NO synthase (eNOS) phosphorylation and NO release. While ACh causes only vasodilation, PAF induces vasoconstriction and hyperpermeability. The key differential signaling mechanisms for discriminating between vasodilation and hyperpermeability are unknown. We tested the hypothesis that differential translocation may serve as a regulatory mechanism of eNOS to determine specific vascular responses. We used ECV-304 cells permanently transfected with eNOS-green fluorescent protein (ECVeNOS-GFP) and demonstrated that the agonists activate eNOS and reproduce their characteristic endothelial permeability effects in these cells. We evaluated eNOS localization by lipid raft analysis and immunofluorescence microscopy. After PAF and ACh, eNOS moves away from caveolae. eNOS distributes both in the plasma membrane and Golgi in control cells. ACh (10(-5) M, 10(-4) M) translocated eNOS preferentially to the trans-Golgi network (TGN) and PAF (10(-7) M) preferentially to the cytosol. We suggest that PAF-induced eNOS translocation preferentially to cytosol reflects a differential signaling mechanism related to changes in permeability, whereas ACh-induced eNOS translocation to the TGN is related to vasodilation.     

Translocation of influenza virus by migrating neutrophils.

Influenza, a predominantly upper respiratory tract infection, replicates in the respiratory epithelia and spreads by an unknown mechanism to the regional lymph nodes. Neutrophils, which accumulate during the early stages of the infection, may be involved in this process. An in vitro model system was used to examine the effect of migrating neutrophils on the permeability of the infected epithelium and on the spread of virus. Epithelial cells (MDCK) infected with influenza virus (WSN H1N1) maintained a stable transepithelial electrical resistance (a measure of epithelial permeability) for 12 hrs. However, when neutrophils migrated across the epithelium toward the virus budding on the apical surface of the epithelium (6 hrs. after infection), the transepithelial electrical resistance fell 24% (P less than 0.001). Neutrophils adhered specifically to the virus and to hemagglutinin expressed exclusively on the apical surface of the cells and phagocytized the free virions. In response to a chemotactic gradient, the infected neutrophils were able to leave the lumenal surface of the infected epithelium, and were able to migrate across the epithelium in equal numbers and at the same rate as uninfected neutrophils. Migration across infected monolayers from the lumenal to the ablumenal surface also caused a fall in resistance (21%, P less than 0.01). Electron microscopic examination of emigrating neutrophils revealed that the leukocytes transported the influenza virions within phagocytic vacuoles and on their surface to the ablumenal side of the monolayer. The results of these studies suggest that the passage of leukocytes across influenza-infected epithelia increases the permeability of the epithelium and provides a route for viral spread.     

我国玉米自交系茎秆性状多样性分析

研究我国玉米自交系茎秆性状特征及其多样性,是培育宜机收玉米品种的重要前提。本研究以兰卡斯特、PB、四平头、旅大红骨和瑞德五大主要类群70份主要玉米自交系为材料,调查12个茎秆相关性状(茎高、穗位高、穗位系数、茎节数、穗位节、穗节系数、穗茎长、穗茎粗、茎鲜重、茎干重、含糖量和含水量),分析性状相关性和类群多样性。结果表明,我国地方种质四平头和旅大红骨茎秆性状表型变异丰富;灌浆期玉米茎秆含水量比较稳定;玉米植株高度与茎节长度显著相关;玉米雌、雄穗节之间的节间数比较恒定;玉米茎秆含糖量与茎节长度、茎粗、果穗着生位置有关;有效降低穗位高度应从降低果穗着生节入手;类群茎秆特征鲜明:兰卡斯特茎节较少,瑞德茎秆较粗,PB茎秆较细,旅大红骨茎秆较粗、茎节较短,四平头植株较矮、茎秆含糖量较低、干物质含量较低;兰卡斯特×四平头和兰卡斯特×PB类群间存在较强的生物量及籽粒产量杂种优势;挖掘和利用茎节较长、穗位较低的玉米地方种质是我国宜机收玉米育种的技术途径。本研究结果对玉米育种具有重要指导意义。