Ionic mechanisms controlling behavioral responses of paramecium to mechanical stimulation

A mechanical stimulus applied to the anterior part of Paramecium causes a transient increase in membrane permeability to calcium. This permits a calcium current to flow into the cell, causing the membrane potential to approach the equilibrium level for calcium. The transient depolarization which results elicits a reversal in the direction of ciliary beat. When the organisms are free-swimming this is seen as the reversed locomotion of Jennings' "avoiding reaction." In contrast, a mechanical stimulus applied to the posterior part results in increased permeability to potassium ions, and hence an outward potassium current. The hyperpolarization which results causes an increase in the frequency of ciliary beat in the normal direction. In free-swimming specimens this is seen as an increase in the velocity of forward locomotion.     

Cortical flow in animal cells

A concerted flow of actin filaments associated with the inner face of the plasma membrane may provide the basis for many animal cell movements. The flow is driven by gradients of tension in the cell cortex, which pull cortical components from regions of relaxation to regions of contraction. In some cases cortical components return through the cytoplasm to establish a continuous cycle. This cortically located motor may drive cell locomotion, growth cone migration, the capping of antigens on a lymphocyte surface, and cytokinesis.     

Wnt5a control of cell polarity and directional movement by polarized redistribution of adhesion receptors

Mechanisms by which Wnt pathways integrate the organization of receptors, organelles, and cytoskeletal proteins to confer cell polarity and directional cell movement are incompletely understood. We show that acute responses to Wnt5a involve recruitment of actin, myosin IIB, Frizzled 3, and melanoma cell adhesion molecule into an intracellular structure in a melanoma cell line. In the presence of a chemokine gradient, this Wnt-mediated receptor-actin-myosin polarity (W-RAMP) structure accumulates asymmetrically at the cell periphery, where it triggers membrane contractility and nuclear movement in the direction of membrane retraction. The process requires endosome trafficking, is associated with multivesicular bodies, and is regulated by Wnt5a through the small guanosine triphosphatases Rab4 and RhoB. Thus, cell-autonomous mechanisms allow Wnt5a to control cell orientation, polarity, and directional movement in response to positional cues from chemokine gradients.     

Rapid actin transport during cell protrusion

Transformed rat fibroblasts expressing two variants of green fluorescent protein, each fused to beta-actin, were used to study actin dynamics during cell protrusion. The recently developed FLAP (fluorescence localization after photobleaching) method permits the tracking of one fluorophore after localized photobleaching by using the other as a colocalized reference. Here, by visualizing the ratio of bleached to total molecules, we found that actin was delivered to protruding zones of the leading edge of the cell at speeds that exceeded 5 micrometers per second. Monte Carlo modeling confirmed that this flow cannot be explained by diffusion and may involve active transport.     

Integrins regulate Rac targeting by internalization of membrane domains

Translocation of the small GTP-binding protein Rac1 to the cell plasma membrane is essential for activating downstream effectors and requires integrin-mediated adhesion of cells to extracellular matrix. We report that active Rac1 binds preferentially to low-density, cholesterol-rich membranes, and specificity is determined at least in part by membrane lipids. Cell detachment triggered internalization of plasma membrane cholesterol and lipid raft markers. Preventing internalization maintained Rac1 membrane targeting and effector activation in nonadherent cells. Regulation of lipid rafts by integrin signals may regulate the location of membrane domains such as lipid rafts and thereby control domain-specific signaling events in anchorage-dependent cells.     

Quantitative imaging of lateral ErbB1 receptor signal propagation in the plasma membrane

Evidence for a new signaling mechanism consisting of ligand-independent lateral propagation of receptor activation in the plasma membrane is presented. We visualized the phosphorylation of green fluorescent protein (GFP)-tagged ErbB1 (ErbB1-GFP) receptors in cells focally stimulated with epidermal growth factor (EGF) covalently attached to beads. This was achieved by quantitative imaging of protein reaction states in cells by fluorescence resonance energy transfer (FRET) with global analysis of fluorescence lifetime imaging microscopy (FLIM) data. The rapid and extensive propagation of receptor phosphorylation over the entire cell after focal stimulation demonstrates a signaling wave at the plasma membrane resulting in full activation of all receptors.     

甘蔗叶绿体荧光参数、MDA含量及膜透性与耐旱性的关系

水分胁迫加速蔗叶活性氧产生并削弱蔗叶活性氧清除能力;随水分胁迫强度的加剧,膜脂过氧化产物丙二醛（ Ｍ Ｄ Ａ）含量迅速提高,细胞质膜透性增大,蔗叶叶绿体２,６ 二氯酚靛酚光还原活性下降,蔗叶叶绿体光系统Ⅱ原初光能转换效率、 Ｐ ＳⅡ潜在活性、叶绿体光合量子产额降低．上述荧光参数的受抑程度与品种的相对耐旱性强弱、膜脂过氧化作用产物 Ｍ Ｄ Ａ 含量增加及质膜相对透性提高程度密切相关     

苎麻生长素结合蛋白BnABP1与GFP融合在烟草中的表达

运用已克隆的苎麻生长素结合蛋白BnABP1基因cDNA及植物表达载体pCAMBIA1300 GFP,构建了35S启动子控制的苎麻BnABP1基因编码区段与绿色荧光蛋白(GFP)基因融合表达重组体(pCAMBIA1300 GFP BnABP1)。通过根癌农杆菌介导法将其转化烟草WS38,经抗性筛选和PCR检测获得了转基因烟草。对转基因烟草细胞进行荧光显微镜观察,发现在细胞质膜和内膜系统上都有较强烈的荧光信号,进一步证明苎麻生长素结合蛋白ABP1已结合在细胞的膜系统上。     

平面叶栅轴流涡轮设计冲角的确定

通过分析最高效率点的实际液流平均进口方向,提出了流道中线决定实际设计的液流平均流动方向的观点,分析了实际机械性能与设计存在偏差的原因,较好解决了涡轮叶片在传统设计过程中冲角选择的问题,从而提高了涡轮设计的正确性,其结论对其他同类涡轮机械的叶片造型设计也具有很好的参考价值。     

Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells

Many proteins associated with the plasma membrane are known to partition into submicroscopic sphingolipid- and cholesterol-rich domains called lipid rafts, but the determinants dictating this segregation of proteins in the membrane are poorly understood. We suppressed the tendency of Aequorea fluorescent proteins to dimerize and targeted these variants to the plasma membrane using several different types of lipid anchors. Fluorescence resonance energy transfer measurements in living cells revealed that acyl but not prenyl modifications promote clustering in lipid rafts. Thus the nature of the lipid anchor on a protein is sufficient to determine submicroscopic localization within the plasma membrane.     

硼对作物细胞膜功能影响的研究进展

综述了硼对植物细胞膜功能的影响机理，指出硼同根细胞膜对硼的吸收密切相关。敏感基因型作物对硼的大量积累和耐硼毒基因型作物对硼的少量积累，都与细胞膜的透性不同有关，并且还与细胞膜和细胞壁的组成相关。硼酸进入根皮细胞可能有两种不同的方式，通过细胞质膜磷脂双分子层的扩散和通过通道蛋白。硼高效品种和低效品种在硼的吸收、运输和再利用方面的差异是细胞膜的组成、细胞膜对硼的透性、膜通道蛋白及其转运效率差异的反映。     

硅对盐胁迫下黄瓜幼苗细胞膜伤害及其保护酶活性的影响

以抗盐性不同的两个黄瓜品种为材料,研究硅对NaCl胁迫下黄瓜幼苗细胞膜伤害及其保护酶活性的影响。结果表明:盐胁迫下细胞的膜质过氧化程度明显加剧,丙二醛(MDA)含量显著增加;超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)的活性均有不同程度的下降。外源硅降低了幼苗叶片质膜透性,提高了盐胁迫下黄瓜幼苗体内SOD、POD酶活性,但对CAT活性影响不大。     

Terrestrial and aquatic orientation in the starhead topminnow, fundulus notti

Starhead topminnows from various shores of a small woodland pond were displaced to unfamiliar surroundings, and their orientation was tested in aquatic and terrestrial arenas. These fish used a sun compass to move in a direction which, at the location of their capture, would have returned them to the land-water interface. The fish accomplished directional terrestrial locomotion by using the position of the sun to align its body for each jump. On heavily overcast days many fish were unable to orient their bodies in a consistent direction from, jump to jump; this inability to orient resulted in random rather than linear movement. There was considerable individual variation in terrestrial locomotor ability.     

高尔基体驻膜糖蛋白GP73启动子克隆

[目的]寻找并克隆有活性的高尔基体膜蛋白GP73的启动子。[方法]对GP73基因转录起始位点上游1 000 bp至下游400 bp序列进行软件分析预测,以肝癌细胞系Huh7基因组DNA为模板,扩增目标片段,构建增强型绿色荧光蛋白(EGFP)为报告基因的重组质粒,转染细胞后在荧光显微镜下观察EGFP的表达,并采用流式细胞仪定量检测转染细胞的荧光强度。[结果]发现GP73转录起始位点上游980到下游330 bp长1 310 bp的序列具有启动子功能。该区域可能具有两个核心启动子序列和多个保守序列,包括TATA box和NF-κB、AP1、GC-SP1等DNA结合序列。[结论]该研究为探讨GP73的转录机制提供了参考。     

高氧气调包装对宰后猪肉蛋白质氧化、钙蛋白酶活性及蛋白质降解的影响

【目的】研究高氧气调包装（HiOx,80% O₂/20% CO₂）对宰后猪肉蛋白质氧化、钙蛋白酶活性及蛋白质降解的影响,探讨高氧气调包装影响猪肉品质的内在机制。【方法】选取12条冷却（4℃）24 h后的杜洛克×长白×约克夏三元杂交猪背最长肌,分别进行高氧气调包装和真空包装（VP）,4℃冷库贮藏,分别在1、4、6 d测定羰基含量及分布、巯基含量、肌节变化、钙蛋白酶活性、肌联蛋白及肌钙蛋白-T降解变化。【结果】高氧气调包装组羰基含量高于真空包装组且贮藏第4和6天差异显著（P＜0.05）。贮藏第1和4天,高氧气调包装组肌细胞外围出现羰基氧化荧光信号,荧光以靠近细胞膜处密度更高,并且逐渐向细胞内部扩散;贮藏第6天,高氧气调包装组细胞膜呈高亮荧光圈,胞内荧光增强,而真空包装组荧光信号较弱。贮藏第6天,高氧气调包装组巯基含量显著低于真空包装组（P＜0.05）。真空包装组宰后肌节M线弱化、A带模糊、肌原纤维Z线断裂;高氧气调包装组肌节结构相对完整。高氧气调包装组在贮藏第1天钙蛋白酶活性显著低于真空包装组（P＜0.05）;高氧气调包装抑制了肌联蛋白和肌钙蛋白-T的降解,且在贮藏第4和6天差异显著（P＜0.05）。【结论】高氧气调包装能够显著提高宰后猪肉蛋白质氧化程度,抑制钙蛋白酶活性发挥及其底物蛋白质的降解。     

Structure of MsbA from E. coli: a homolog of the multidrug resistance ATP binding cassette (ABC) transporters

Multidrug resistance (MDR) is a serious medical problem and presents a major challenge to the treatment of disease and the development of novel therapeutics. ABC transporters that are associated with multidrug resistance (MDR-ABC transporters) translocate hydrophobic drugs and lipids from the inner to the outer leaflet of the cell membrane. To better elucidate the structural basis for the "flip-flop" mechanism of substrate movement across the lipid bilayer, we have determined the structure of the lipid flippase MsbA from Escherichia coli by x-ray crystallography to a resolution of 4.5 angstroms. MsbA is organized as a homodimer with each subunit containing six transmembrane alpha-helices and a nucleotide-binding domain. The asymmetric distribution of charged residues lining a central chamber suggests a general mechanism for the translocation of substrate by MsbA and other MDR-ABC transporters. The structure of MsbA can serve as a model for the MDR-ABC transporters that confer multidrug resistance to cancer cells and infectious microorganisms.     

Imaging intracellular fluorescent proteins at nanometer resolution

We introduce a method for optically imaging intracellular proteins at nanometer spatial resolution. Numerous sparse subsets of photoactivatable fluorescent protein molecules were activated, localized (to approximately 2 to 25 nanometers), and then bleached. The aggregate position information from all subsets was then assembled into a superresolution image. We used this method--termed photoactivated localization microscopy--to image specific target proteins in thin sections of lysosomes and mitochondria; in fixed whole cells, we imaged vinculin at focal adhesions, actin within a lamellipodium, and the distribution of the retroviral protein Gag at the plasma membrane.     

The casparian strip as a barrier to the movement of lanthanum in corn roots

The effectiveness of the Casparian strip as a barrier to apoplastic movement of solutes from cortex to stele of corn roots was investigated by using lanthanum in combination with electron microscopy. Lanthanum deposits were found only in cell walls and on the outside of the plasma membrane of epidermal, cortical, and endodermal cells up to the Casparian strip. Lanthanum was completely absent from the stele, indicating that the Casparian strip provides an effective barrier to apoplastic movement of solutes. Inhibitory effects of trivalent lanthanum ions on the absorption of potassium ions are discussed in relation to the nature of the lanthanum ion binding site on membranes.     

冷等离子体对单核增生李斯特菌的杀菌机理

【目的】作为一种新兴的非热灭菌技术,冷等离子在食品行业中得到了广泛的应用。通过探讨冷等离子体对细菌细胞膜的破坏效果,阐述其抗菌机制,为冷等离子体在食品行业的应用提供参考。【方法】本研究以单核增生李斯特菌(Listeria monocytogenes,LM)为试验菌株,研究冷等离子体处理对LM形态和胞内物质的影响,阐述LM细胞膜完整性的变化;冷等离子体处理后,通过测定细胞膜脂肪酸含量和类型的变化,比较8-苯胺-1-萘磺酸荧光强度的改变,反映冷等离子体对LM细胞膜流动性的影响。通过测定碘化丙啶荧光、电导率和β-半乳糖苷酶活性的变化,观察冷等离子体对LM细胞膜通透性的改变。最后,通过检测胞内活性氧和活性氧相关基因表达量的变化,阐明冷等离子体对LM细胞膜造成的氧化损伤。【结果】冷等离子体处理后,LM细胞膜表面观察到破损变形的结构,胞内蛋白质和DNA分别下降了68 mg·mL^-1和14μg·mL^-1,证明冷等离子体破坏了细胞膜的完整性。冷等离子体处理使LM细胞膜中不饱和脂肪酸的含量从40.17%上升至53.91%,饱和脂肪酸的含量从53.68%下降到4...     

Visual receptor potential: modification by injected current in the limulus lateral eye

The latent period of the light-evoked receptor potential was increased by hyperpolarizing currents injected directly into doubly impaled retinular cells. Indirect hyperpolarization of these cells by injection of hyperpolarizing current into the eccentric cell or other intraommatidial retinular cells either shortened or did not change the latent period. The modification of the latent period may depend upon the direction of current flow across some regions of the membrane system constituting the rhabdomere. The reduction in magnitude of the receptor potential obtained with strong hyperpolarizing currents may also depend upon the direction of current flow. The results support the conclusion that the receptor potential originates in retinular cells within the membrane system of the rhabdomere.