Ouabain resistance conferred by expression of the cDNA for a murine Na+, K+-ATPase alpha subunit

The molecular basis for the marked difference between primate and rodent cells in sensitivity to the cardiac glycoside ouabain has been established by genetic techniques. A complementary DNA encoding the entire alpha 1 subunit of the mouse Na+- and K+-dependent adenosine triphosphatase (ATPase) was inserted into the expression vector pSV2. This engineered DNA molecule confers resistance against 10(-4) M ouabain to monkey CV-1 cells. Deletion of sequences encoding the carboxyl terminus of the alpha 1 subunit abolish the activity of the complementary DNA. The ability to assay the biological activity of this ATPase in a transfection protocol permits the application of molecular genetic techniques to the analysis of structure-function relationships for the enzyme that establishes the internal Na+/K+ environment of most animal cells. The full-length alpha 1 subunit complementary DNA will also be useful as a dominant selectable marker for somatic cell genetic studies utilizing ouabain-sensitive cells.     

Isoform-specific modulation of Na+, K+-ATPase alpha-subunit gene expression in hypertension

Sodium, potassium-adenosine triphosphatase (Na+, K+-ATPase) is hypothesized to be involved in systemic vascular hypertension through its effects on smooth muscle reactivity and myocardial contractility. By means of RNA blot analyses of cardiac, aortic, and skeletal muscle RNAs in two rat hypertensive models, Na+,K+-ATPase alpha-subunit messenger RNA isoforms (alpha 2 and alpha 3) were shown to be deinduced in response to increased intravascular pressure. The changes were observed after 48 hours or more of experimental hypertension. Under these conditions, there is coordinate induction of another alpha isoform (alpha 1) and of beta-subunit messenger RNAs, probably in response to alterations in sodium flux rather than to elevated blood pressure.     

女贞子提取物对大鼠不同组织Na~+,K~+-ATPase活性的影响

通过女贞子提取物对大强度耐力训练大鼠不同组织Na+,K+-ATPase活性的影响,探讨女贞子提取物对大鼠运动能力的作用机制。以24只SD大鼠随机分为安静对照组、运动对照组和运动+女贞子组,每组8只。运动对照组进行6周大强度跑台训练,运动+女贞子组除大强度跑台训练外,每天灌服2 mL浓度为400 mg.kg-1女贞子提取物。安静对照组和运动对照组灌胃同体积0.5%Tween-80溶液。6周后安静对照组在安静时、运动对照组和运动+女贞子组进行一次力竭性运动后取材,测定不同组织Na+,K+-ATPase活性。结果:运动对照组和运动+女贞子组大鼠各组织Na+,K+-ATPase活性均显著低于安静对照组,运动+女贞子组大鼠不同组织Na+,K+-ATPase活性显著高于运动对照组;运动+女贞子组大鼠力竭运动时间比运动对照组延长23.09%。结论:补充女贞子提取物可以调节大鼠不同组织Na+,K+-ATPase活性,增加Na+,K+-ATPase活性,延长运动至疲劳的时间。     

T-cell recognition of Ia molecules selectively altered by a single amino acid substitution

T lymphocytes recognize foreign antigen together with allele-specific determinants on membrane-bound class I and class II (Ia) gene products of the major histocompatibility complex. To identify amino acids of class II molecules critical to this recognition process, the genes encoding the beta chains of the I-Ak molecule were cloned from a wild-type B-cell hybridoma and from an immunoselected variant subline showing distinct serological and T-cell stimulatory properties. Nucleotide sequencing and DNA-mediated gene transfer established that a single base transition (G----A) encoding a change from glutamic acid to lysine at position 67 in the I-Ak beta molecule accounted for all the observed phenotypic changes of the variant cells. These results confirm the importance of residues 62 to 78 in the amino terminal domain of I-A beta for class II-restricted T-cell recognition of antigen and demonstrate the ability of a single substitution in this region to alter this recognition event.     

松油烯-4-醇及其酯类衍生物对家蝇Na+，K+-ATPase活性的抑制作用

【目的】探讨松油烯-4-醇的杀虫作用机理。【方法】采用点滴法测定了松油烯-4-醇及其6种酯类衍生物乙酸酯（Z1）、丙酸酯（Z2）、丙烯酸酯（Z3）、苯甲酸酯（Z4）、苯磺酸酯（Z5）、对甲苯磺酸酯（Z6）对家蝇成虫的触杀活性及对其体内Na+,K+-ATPase活性的影响。【结果】松油烯-4-醇及其酯类衍生物对家蝇均具有一定的触杀活性,除丙烯酸酯外,其它酯类衍生物的触杀毒力均高于或相当于松油烯-4-醇;活体条件下,7种化合物均对Na+,K+-ATPase具有较强的抑制作用,且随着中毒症状的加剧,对该酶的抑制作用增强;离体条件下,松油烯-4-醇及其乙酸酯、丙酸酯、苯甲酸酯、苯磺酸酯、对甲苯磺酸酯衍生物对家蝇Na+,K+-ATPase的IC50分别为155.89、197.98、96.02、121.36、124.85、153.74μg•ml-1;分析发现松油烯-4-醇及其酯类衍生物对家蝇24 h的触杀LD50与对Na+,K+-ATPase的IC50之间具有显著相关性。【结论】Na+,K+-ATPase可能是松油烯-4-醇类化合物的主要作用靶标。     

Inhibition of Na＋,K＋-ATPase in Housefly （Musca domestica L.） by Terpinen- 4-ol and Its Ester Derivatives

To reveal the insecticidal mechanism of terpinen-4-ol, the activity of Na＋,K＋-ATPase in insects tested were determined in vivo and in vitro. The results showed that terpinen-4-ol and its ester derivatives had strong contact activity to housefly and the contact toxicities of its derivatives except Z3 were all superior or equivalent to terpinen-4-ol. All the 7 compounds had strong inhibition towards activity of Na＋,K＋-ATPase. With poisoning symptom exacerbating, the inhibition rates were gradually increased. In vitro, the IC50 of terpinen-4-ol, Z1, Z2, Z4, Z5, and Z6 was 155.89, 197.98, 96.02, 121.36, 124.85, and 153.74 μg mL% respectively. There was well correlation between the LDs0 of terpinen-4-ol derivatives to housefly and the IC50 of terpinen-4-ol derivatives to Na＋,K＋-ATPase in housefly. In conclusion, Na＋,K＋-ATPase was likely the target of terpinen-4-ol against insects.     

Widespread occurrence in frogs and toads of skin compounds interacting with the ouabain site of Na+, K+-ATPase

Amphibians of the family Bufonidae contain high levels of skin compounds that both inhibit Na+- and K+-dependent adenosinetriphosphatase and antagonize the binding of ouabain to the enzyme. In species of Bufo and Atelopus, these compounds are relatively nonpolar bufodienolides, whereas Dendrophryniscus and Melanophryniscus contain more polar compounds of unknown structure. Skin extracts from 30 of 48 species of frogs representing an additional eight families contained relatively low levels of compounds that inhibit binding of ouabain to Na+,K+-adenosinetriphosphatase. The widespread occurrence of low levels of inhibitory compounds is consonant with the role for these compounds as physiological regulators of Na+,K+-adenosinetriphosphatase in amphibian skin; high levels in the Bufonidae probably also serve as a defense against some predators.     

盐胁迫对香蕉幼苗体内Na^＋和K^＋含量及其分布的影响

采用营养液栽培,以香蕉幼苗为试材,研究了盐胁迫对香蕉幼苗根部、假茎和叶片各部位Na^＋和K^＋含量以及K＋／Na^＋的影响。结果表明,与对照相比植株体内Na^＋的含量增加,而植物体内K^＋含量减少,K^＋／Na^＋下降显著。     

NaCl胁迫对不同南瓜品种幼苗Na+、K+量的影响

采用19个不同类型南瓜品种,研究了300mmol/L NaCl下,幼苗地上部和根系Na+、K+离子含量、Na+/K+比值的变化.结果表明,NaCl胁迫8d后,不同南瓜品种幼苗Na+含量均明显增加,而K+含量下降,离子平衡被打破.南瓜幼苗体内的Na+含量、地上部的Na+/K+比值的变化反映植物对盐离子和营养元素相对的吸收情况.青栗(Q1)南瓜幼苗根系Na+含量、地上部Na+/K+比值明显高于黑蜜南瓜(H2)和黑子南瓜(H3).不同南瓜品种幼苗体内Na+含量、地上部Na+/K+比值变化趋势与NaCl胁迫下不同南瓜品种幼苗的盐害指数的结果基本一致.进一步验证了Q1耐盐性强与盐胁迫下地上部Na+/K+比值较低,K+离子含量较高有关;而品种H2和H3对盐敏感与盐胁迫下地上部Na+/K+比值较高,K+离子含量较低有关.     

不同盐碱条件下白榆器官中K~+、Na~+、Ca~(2+)和Mg~(2+)分布特征

为了探明白榆(Ulmus pumila)适应盐碱环境的机理,以高盐、中低盐、非盐碱3个土壤盐分立地条件下生长的白榆为试验材料,研究不同盐分浓度下新叶、老叶、新枝、老枝和根5个营养器官中4种金属盐离子分布特征.结果表明,白榆体内K 、Na 、Ca2 和Mg2 不同部位的含量不同,并存在显著差异;白榆各部位K 、Na 含量随着生长地区土壤盐分的增加而增加;K /Na 幼嫩器官显著高于成熟器官,茎叶显著高于根部;含盐中低等土壤中生长的白榆K /Na 高于高盐和非盐碱土壤中生长白榆.白榆器官中K 、Na 、Ca2 对盐碱的敏感性高于Mg2 ,K 和Na 是4种金属盐离子中白榆在盐渍生境下进行渗透调节作用的主要因素.     

盐胁迫对蓖麻Na+、K+吸收分布和叶绿素荧光的影响

[目的]探讨蓖麻的耐盐机制。[方法]比较了栽培蓖麻淄蓖6号和天然生长于滨海盐碱地的野生蓖麻体内K+、Na+的分布特点、叶绿素荧光参数、植株生长状况和叶片渗透调节能力对NaC l处理的响应。[结果]野生蓖麻较栽培蓖麻具更明显的喜盐特性;蓖麻的耐盐机制在于促进K+的吸收和向地上部的运输,进而维持叶片的K+/Na+平衡;通过维持光系统Ⅱ的稳定性进而维持盐胁迫下叶片的光合速率。[结论]该研究为蓖麻在盐碱滩涂地的推广应用提供了理论依据。     

盐胁迫对蓖麻Na~+、K~+吸收分布特点和叶绿素荧光的影响(英文)

[Objective] The aim of this study is to reveal the salt resistance of castor. [Method] Under salt stress, the growth, osmotic potential, chlorophyll fluorescence parameters, Na~+ and K~+ uptakes and transports in the seedlings of two Ricinus communis varieties (cultivar castor ‘Zibi 6’ and wild castor which grew naturally in coastal saline-alkali land), were comparatively studied. [Result] Wild castor preformed better in halophilism than that of cultivar castor Zibi 6 under the NaCl treatment. One of the salt tolerant mechanisms of castor is to improve K~+ uptake and transport to overground portion, thus to maintain K~+/Na~+ homeostasis in leaves; on the other hand, the high stability of Photoreaction System Ⅱ (PS Ⅱ) plays a key role in maintaining the leaf photosynthetic rate under salt stress. [Conclusion] The results of this study provided theoretical basis for the extension and application of castor in saline beach.     

钠、钾和钙离子对小麦幼苗叶片质膜透性和组织活力的效应

在等渗和等阴离子有效浓度条件下,Ｎａ＾＋和Ｋ＾＋的单效效应都使小麦幼苗叶片的质膜透性增大,且Ｎａ＾＋〉Ｋ＾＋,而Ｃａ＾２＋的单独效应却使其减小;Ｋ＾＋和Ｃａ＾２＋的单独效应都使小麦叶片组织ＴＴＣ还原率增大,且Ｃａ＾２＋〉Ｋ＾＋,而Ｎａ＾＋的单独效应却使其减小。     

喹乙醇对鲤鳃组织Na+、K+-ATP酶活性及血浆生化指标的影响

用含不同喹乙醇剂量的饲料饲喂健康鲤鱼种,研究饲料中不同喹乙醇剂量对鲤体成分含量、血浆生化指标以及鲤鳃组织．Na^ 、K^ -ATP酶活性的影响,探讨喹乙醇对鲤的生化毒性。试验分成6个喹乙醇处理组(0、200、400、800、1600、3200mg／kg),试验期60d。试验结束时分别从各组中采血、取全鱼样和鱼鳃样,进行不同指标测试。结果表明,喹乙醇使鱼体脂肪含量轻度上升,使灰分沉积量显著降低,并呈现剂量一反应关系。与对照组比较,在较高喹乙醇剂量下,谷草转氨酶(AST)、肌酸激酶(CK)活性降幅较大,血浆中葡萄糖(GLU)、胆固醇(CHO)、总胆红素(T-B)、直接胆红素(D-B)、高密度脂蛋白(HDL)、白蛋白(ALB)、K^ 、Ca^2 、P含量升高,肌酐(CRE)和CO2含量降低,其他测定指标变化不明显。鳃组织中Na^ 、K^ -ATP酶活性随喹乙醇剂量的升高而呈逐渐下降的趋势,呈现剂量效应关系,喹乙醇剂量达1600mg／k以上时,Na^ 、K^ -ATP酶活性显著降低。结论：1600mg／k以上的喹乙醇剂量可以明显干扰鲤的正常生化代谢功能。     

Alteration of four identified K+ currents in Drosophila muscle by mutations in eag

Voltage-clamp analysis of Drosophila larval muscle revealed that ether à go-go (eag) mutations affected all identified potassium currents, including those specifically eliminated by mutations in the Shaker or slowpoke gene. Together with DNA sequence analysis, the results suggest that the eag locus encodes a subunit common to different potassium channels. Thus, combinatorial assembly of polypeptides from different genes may contribute to potassium channel diversity.     

Conversion of a PI-anchored protein to an integral membrane protein by a single amino acid mutation

Qa-2, a cell-surface glycoprotein anchored by phosphatidylinositol (PI), is structurally related to the class I transplantation antigens H-2 K, D, and L, which are integral membrane glycoproteins. The predicted transmembrane segment of Qa-2 differs from those of H-2 K, D, and L by the presence of an aspartate in place of a valine at position 295. A single base change that replaced this aspartate with valine resulted in cell-surface Qa-2 molecules that were insensitive to hydrolysis by a PI-specific phospholipase C and more resistant to papain cleavage, properties shared by H-2D. Cells expressing Asp----Val mutant Qa-2 proteins were still able to attach a PI anchor to endogenous proteins such as Thy-1 and J11D. It therefore appears that this single amino acid change converts Qa-2 from a PI-linked form into an integral membrane protein.     

NaCl对咸海卡拉白鱼血清离子、皮质醇和组织Na+,K+-ATP酶活性的影响

将卡拉白鱼(Chalcalburnus chalcoides aralensis)在分别含NaCl 0,2,4,6,8 g/L的水体中饲养30 d后发现,NaCl质量浓度2 g/L和4 g/L组鱼相对增重率与对照组相比无显著性差异,生长良好;6 g/L和8 g/L组鱼相对增重率低于对照组(P<0.05),生长受抑制。血清渗透压随盐度升高而增高(P>0.05);NaCl 4,6,8 g/L组血的Na ,Cl-浓度高于对照组(P<0.05);6 g/L和8 g/L组血K 浓度低于对照组(P<0.05);鳃Na ,K -ATP酶活性除8 g/L组下降外,其余各组鱼均高于对照(P<0.05);血清皮质醇水平随NaCl的升高而呈线性增加的趋势(P>0.05)。     

盐胁迫对芦笋幼苗生长和体内 Na＋，K＋，Ca2＋分布的影响

以芦笋品种 NJ978为试材,研究了 NaCl 胁迫对芦笋幼苗生长及体内 Na ＋,K ＋,Ca2＋吸收和分布的影响,结果表明：盐分对芦笋幼苗生长的抑制作用随 NaCl 浓度的增加而加剧,低盐胁迫下（NaCl ≤50 mmol/L ）,芦笋生长与对照没有显著差异,高盐胁迫（200～300 mmol/L NaCl）显著抑制了幼苗生长。随 NaCl 浓度的增加,芦笋体内Na ＋含量增加,K ＋,Ca2＋含量降低。芦笋根系对 Na ＋有一定的截留能力,低盐环境下（NaCl ≤100 mmol/L ）,根部Na ＋迅速增加,而地上部 Na ＋增加缓慢,从而抑制了 Na ＋向地上部运输,并维持了地上部相对稳定的 K ＋,Ca2＋平衡;外界 NaCl 浓度高于根系 Na ＋截留阈值后（100 mmol/L）,大量的 Na ＋运输至地上部并限制了 K ＋,Ca2＋的吸收,表现为 K ＋/Na ＋和 Ca2＋/Na ＋迅速下降。芦笋根系对 K ＋,Ca2＋,Na ＋选择吸收性（ASK ,Na ,ASCa ,Na ）随盐胁迫增加而变高;根部向地上部运输 K ＋,Ca2＋能力（TSK ,Na ,TSCa ,Na ）在0～100 mmol/L NaCl 水平下逐渐升高,之后迅速下降。以上结果表明：根部对 Na ＋的截留和 Na ＋,K ＋,Ca2＋在器官水平上的再分布是芦笋适应盐胁迫的重要机制之一。     

H_2O_2预处理对盐胁迫下黄瓜幼苗生长和K~+,Na~+,Cl~-分布的影响

以黄瓜品种津春2号为试材,采用营养液水培,研究了H2O2预处理对NaCl胁迫下幼苗植株不同器官中K+,Na+和Cl-含量的影响。结果表明,NaCl胁迫下黄瓜植株体内K+含量下降、Na+和Cl-含量升高、K/Na和Cl/Na比值降低;H2O2预处理抑制了盐胁迫下黄瓜幼苗植株体内K+含量的降低,减少了Na+和Cl-在叶片的积累,提高了K+,Na+吸收和运输的选择性及K/Na比值,缓解了盐胁迫的伤害,改善了盐胁迫下黄瓜幼苗的生长,显著提高了根、茎、叶的干重和幼苗株高。说明,叶面喷施外源H2O2可通过调控盐胁迫下植株体内离子在器官水平的区域化分布,进而提高黄瓜幼苗的耐盐性。     

混合药剂吸湿回干技术对盐碱胁迫大豆苗期植株K~+、Ca~(2+)和Na~+的影响

为明确不同处理对大豆苗期耐盐碱能力的影响,对大豆种子采取引发剂处理、水处理和未处理三种处理方式,研究了盐碱胁迫下苗期植株K~+、Ca~(2+)和Na~+的变化。结果表明:种子引发剂处理的苗期植株,在盐碱胁迫下其地上部和地下部Na~+、K~+和Ca~(2+)含量均高于未处理和水处理;引发剂处理的TSK,Na最高,而其TSCa,Na高于水处理,低于未处理。通过混合药剂吸湿回干技术处理,可以增强大豆种子的抗盐碱性,为解决大豆耐盐碱能力提供新的理论依据。