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1.
拟除虫菊酯对家蝇Na-K-ATPase抑制作用的研究 总被引:16,自引:6,他引:16
通过对家蝇神经系统Na-K-ATPase性质的研究,表明Na-K-ATPase反应的适宜pH值为7.0~8.0,适温为35~40℃,Km为0.22 mmol/L,Vmax为555.56 nmol/(mg·min)。比较测定了家蝇敏感品系、Del-R、2Cl-R抗性品系的Na-K-ATPase活性及溴氰菊酯和氯菊酯对该酶的抑制作用。实验证明,敏感与抗性品系间Na-K-ATPase活力无显著的差异,但溴氰菊酯和氯菊酯对不同家蝇品系Na-K-ATPase的抑制作用有明显区别,两种拟除虫菊酯可抑制敏感家蝇品系Na-K-ATPase的活性,而对抗性品系无明显的抑制作用。 相似文献
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测定敏感、抗溴氰菊酯(Del-R)、抗氯菊酯(2Cl-R)的家蝇品系对有机磷杀虫剂敌敌畏、辛硫磷及马拉硫磷的LD50,α-乙酸萘酯(α-NA)酯酶动力学,酯酶的活性和酯酶的抑制作用。Del-R和2Cl-R的家蝇品系对三种有机磷杀虫剂的抗性倍数为0.966~7.190倍,均为低抗水平。三个家蝇品系的羧酸酯酶活性水平与抑制中浓度存在正相关性,说明羧酸酯酶在抗拟除虫菊酯家蝇对有机磷杀虫剂的抗性中起一定的作用。 相似文献
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家蝇对拟除虫菊酯农药的抗性机制 总被引:15,自引:8,他引:15
本文对二氯苯醚菊酯和溴氰菊酯分别选择的高抗性家蝇(Musca domestica vicina)品系2Cl-R及Dec-R的抗性机制进行了研究.应用生物测定、增效剂,体内试验的表皮穿透作用、离体条件的解毒酶系活性的增加以及家蝇头部ATP酶活力的研究结果表明,两种拟除虫菊酯高抗性家蝇品系的表皮穿透性均比正常品系NP为慢,特别是Dec-R品系极慢.酯酶和多功能氧化酶及其末端的细胞色素P-450的活性在两个抗性品系中都比NP品系有不同程度的增高,但2Cl-R品系以氧化酶为主,而Dec-R品系似以酯酶占优势.Dec-R品系的Na+—K+-ATP酶活力低于NP品系的46%,而2Cl-R品系与NP品系相等.Mg2+-ATP酶活性在两个抗性品系中均高于正常品系.Mg2+-ATP酶可能也是拟除虫菊酯的一个重要靶标部位. 相似文献
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烟粉虱对拟除虫菊酯杀虫剂的抗性机理 总被引:3,自引:1,他引:3
通过增效剂生物测定、生化分析以及钠离子通道基因ⅡS4-6 cDNA片段的RT-PCR扩增,探讨了烟粉虱Bemisia tabaci(Gennadius)对拟除虫菊酯杀虫剂的抗性机理。结果表明:对于采自田间的6个烟粉虱抗性品系,磷酸三苯酯(TPP)和胡椒基丁醚(PBO)对氯氰菊酯、溴氰菊酯、氯氟氰菊酯和甲氰菊酯均有显著的增效作用,而DEM对4种拟除虫菊酯杀虫剂均无明显的增效作用。烟粉虱抗性品系的α-NA羧酸酯酶和β-NA羧酸酯酶活性分别是敏感品系的2.16~2.65倍和1.22~1.41倍,抗性品系的谷胱甘肽S转移酶活性与敏感品系没有差异,表明羧酸酯酶和多功能氧化酶在烟粉虱对拟除虫菊酯类杀虫剂的抗性中具有重要的作用,而谷胱甘肽S转移酶与抗性无关。通过RT-PCR克隆了6个烟粉虱田间抗性品系的钠离子通道结构域ⅡS4-6 cDNA片段的序列(420 bp),发现与敏感品系相比,有2个位点发生突变,分别为L925I突变和I917V突变,L925I突变在所有6个烟粉虱田间抗性种群中均有发生,该位点突变已被证实与拟除虫菊酯类杀虫剂密切相关,表明神经不敏感性可能是烟粉虱对拟除虫菊酯产生抗性的另一个重要因子。 相似文献
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化学防治是控制蚊虫传播疾病的主要方法, 抗性监测表明我国蚊虫已对有机磷、 有机氯、 氨基甲酸酯和拟除虫菊酯类杀虫剂产生了不同程度的抗性。蚊虫抗药性的分子机制主要包括靶标抗性和三大解毒酶家族带来的代谢抗性。筛选对杀虫剂敏感的品系是抗性监测和抗性机理研究必不可少的材料。本研究通过从一个致倦库蚊Culex pipiens quinquefasciatus野生种群筛选无乙酰胆碱酯酶G119S突变且具有低活性羧酸酯酶、 P450单加氧酶和谷胱甘肽-S-转移酶的单雌系, 建立了一个对杀虫剂敏感的致倦库蚊品系。该品系的羧酸酯酶活性是敏感品系S-lab的2.5倍, P450单加氧酶和谷胱甘肽-S-转移酶的活性与S-lab相当。生物测定表明, 与S-lab相比, 该品系对有机磷杀虫剂有低于2倍的抗性, 对氨基甲酸酯和拟除虫菊酯类杀虫剂没有抗性, 可以作为相对敏感品系用于抗性监测。 相似文献
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云南烟蚜抗药性机制研究 总被引:1,自引:0,他引:1
通过比较云南烟蚜敏感品系和抗性品系的解毒酶(α-乙酸萘酯羧酸酯酶、β-乙酸萘酯羧酸酯酶)和靶标酶(乙酰胆碱酯酶)的活力,研究了烟蚜对有机磷、拟除虫菊酯和氨基甲酸酯类杀虫剂抗性的生化机制,并通过酯酶基因扩增检测和钠离子通道突变检测,研究了其抗性的分子机制。结果表明:α-乙酸萘酯羧酸酯酶活力增强是烟蚜对有机磷类、氨基甲酸酯类杀虫剂及拟除虫菊酯类杀虫剂的抗性机制之一;乙酰胆碱酯酶在烟蚜对有机磷杀虫剂抗性中起重要作用;3个抗性品系烟蚜均没有发生酯酶基因扩增,抗拟除虫菊酯品系烟蚜发生了钠离子通道突变。 相似文献
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家蝇对DDT和拟除虫菊酯的重要抗性机制——中枢神经系统的不敏感性 总被引:4,自引:1,他引:3
本文以电生理技术研究了四个品系的家蝇Musca domestica vicina Macq.中枢神经系统(CNS)对DDT、二氯苯醚菊酯和澳氰菊酯的敏感性,结果表明:三种抗性家蝇,DDT高抗品系(DDT-R)、二氯苯醚菊酯高抗品系(2C1-R)和溴氰菊酯高抗品系(Dec-R)的中枢神经系统(CNS)对三种杀虫剂的敏感性与敏感家蝇相比均明显降低,而且,GNS的不敏感性随杀虫剂LD50的升高有逐渐上升的趋势.我们认为,CNS不敏感性是家蝇对DDT相拟除虫菊酯产生抗性的一个重要机制,也是产生交互抗性的一个重要原因. 相似文献
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利用免疫印迹,免疫电镜和ATP水解活性的测定对豌豆(Pisum sativum L.)根细胞胞质中V1-ATPase复合物的存在进行鉴定。用兔抗绿豆V-typeH^ -ATPase的A,B亚基的抗体进行的immuno-blotting和胶体金电镜结果都表明,胞质中存在有A,B亚基。活性测定结果进一步表明胞质具有ATP水解活性,这些结果说明豌豆根胞质具有活性的V1-ATPase复合物。这是首次直接证明植物中有胞质V1-ATPase的存在。 相似文献
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We report here characterization of calmodulin-stimulated Ca2+ transport activities in synaptic plasma membranes (SPM). The calcium transport activity consists of a Ca2+-stimulated, Mg2+-dependent ATP hydrolysis coupled with ATP-dependent Ca2+ uptake into membraneous sacs on the cytosolic face of the synaptosomal membrane. These transport activities have been found in synaptosomal subfractions to be located primarily in SPM-1 and SPM-2. Both Ca2+-ATPase and ATP-dependent Ca2+ uptake require calmodulin for maximal activity (KCm for ATPase = 60 nM; KCm for uptake = 50 nM). In the reconstituted membrane system, KCa was found to be 0.8 microM for Ca2+-ATPase and 0.4 microM for Ca2+ uptake. These results demonstrate for the first time the calmodulin requirements for the Ca2+ pump in SPM when Ca2+ ATPase and Ca2+ uptake are assayed under functionally coupled conditions. They suggest that calmodulin association with the membrane calcium pump is regulated by the level of free Ca2+ in the cytoplasm. The activation by calmodulin, in turn, regulates the cytosolic Ca2+ levels in a feedback process. These studies expand the calmodulin hypothesis of synaptic transmission to include activation of a high-affinity Ca2+ + Mg2+ ATPase as a regulator for cytosolic Ca2+. 相似文献
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Active transport of lead by human red blood cells 总被引:1,自引:0,他引:1
T J Simons 《FEBS letters》1984,172(2):250-254
Human red cells suspended in lead-citrate buffers (2.6 microM Pb2+) take up much less Pb than predicted from studies of equilibrium binding of Pb to haemolysates. Pb uptake is increased by ATP depletion, or by loading at 0 degrees C. Tracer studies with 203Pb indicate that the low uptake at 37 degrees C in the presence of substrate is not due to membrane impermeability to Pb. Cold-loaded cells extrude Pb against a concentration gradient at 37 degrees C when glucose is present. These results suggest that the cellular loading of Pb is dependent on the balance between an inward leak and an outward pump. The extrusion of Pb from the cells is possibly brought by the Ca pump. 相似文献
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Ca2+ uptake into the endoplasmic reticulum (ER) is mediated by Ca2+ ATPase isoforms, which are all selectively inhibited by nanomolar concentrations of thapsigargin. Using ATP/Mg2+-dependent 45Ca2+ transport in rat brain microsomes, tissue sections, and permeabilized cells, as well as Ca2+ imaging in living cells we distinguish two ER Ca2+ pools in the rat CNS. Nanomolar levels of thapsigargin blocked one component of brain microsomal 45Ca2+ transport, which we designate as the thapsigargin-sensitive pool (TG-S). The remaining component was only inhibited by micromolar thapsigargin, and thus designated as thapsigargin resistant (TG-R). Ca2+ ATPase and [32P]phosphoenzyme assays also distinguished activities with differential sensitivities to thapsigargin. The TG-R Ca2+ uptake displayed unique anion permeabilities, was inhibited by vanadate, but was unaffected by sulfhydryl reduction. Ca2+ sequestered into the TG-R pool could not be released by inositol-1,4,5-trisphosphate, caffeine, or cyclic ADP-ribose. The TG-R Ca2+ pool had a unique anatomical distribution in the brain, with selective enrichment in brainstem and spinal cord structures. Cell lines that expressed high levels of the TG-R pool required micromolar concentrations of thapsigargin to effectively raise cytoplasmic Ca2+ levels. TG-R Ca2+ accumulation represents a distinct Ca2+ buffering pool in specific CNS regions with unique pharmacological sensitivities and anatomical distributions. 相似文献
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急性运动所致线粒体某些功能的改变及胆红素的保护作用 总被引:9,自引:0,他引:9
目的:探讨急性运动所致疲劳的机理以及胆红素的保护作用。方法:将Wistar大鼠随机分为对照组、运动组、运动恢复组、胆红素处理运动组、胆红素处理运动恢复组,共5组,分别灌胃1μmol/Kg体重的胆红素或生理盐水4周,负重(体重的5%)游泳2h后处死,测定有关指标。结果:急性运动后即刻大鼠排肠肌胞、线粒体Ca^2 含量明显升高,恢复12h后线粒体Ca^2 含量呈继续升高的趋势;线粒体C^a2 -Mg^2 -ATP酶、Ca^2 -ATP酶活性明显下降,12h后有所回升,胆红素处理后可以明显抑制这些指标的改变,但线粒体Mg^2 -ATP酶活性的变化在胆红素处理组和未处理组间无明显差异,均明显低于对照组,只是胆红素组的恢复相对较快。结论:生理浓度的胆红素可能通过抑制线粒体某些功能的改变而保护细胞名受急性运动所致的损伤,从而延缓疲劳的发生,加速恢复。 相似文献
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Ca2+预处理对热胁迫下辣椒叶肉细胞中Ca2+-ATP酶活性的影响 总被引:2,自引:0,他引:2
在常温下生长的辣椒(Capsicum
annum L.)叶肉细胞中Ca2+-ATP酶主要分布于质膜、液泡膜上,叶绿体的基质和基粒片层上也有少量分布;在40℃下热胁迫不同的时间,酶活性逐渐下降,直至叶绿体超微结构解体.同样条件下,经过Ca2+预处理后,分布在上述细胞器膜或片层上的酶活性大大提高,表明Ca2+预处理对该酶活性具有激活作用;Ca2+预处理对热胁迫下的超微结构的完整性具有一定的保护作用,并且能使Ca2+-ATP酶在热胁迫下维持较高活性.结果表明,Ca2+预处理增强辣椒幼苗的抗热性,可能与其稳定细胞膜、从而使Ca2+-ATP酶在热胁迫下保持较高活性有一定关系. 相似文献
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The Ca(2+)-ATPase activity of rat brain microsomes was studied in streptozotocin (STZ)-induced diabetes. Male rats, 200-250 g, were rendered diabetic by injection of STZ (45 mg kg(-1) body weight) via the teil vein. Brain tissues were collected at 1, 4 and 10 weeks after diabetes was induced for determination of Ca(2+)-ATPase activity, lipid peroxidation and tissue calcium levels. Diabetic rats had significantly elevated blood glucose levels compared to controls. Blood glucose levels were 92.92 +/- 1.22 mg dl(-1) (mean +/- SEM) for the control group, 362.50 +/- 9.61 mg dl(-1) at 1 week and >500 mg dl(-1) at 4, 8 and 10 weeks for the diabetics. Enzyme activities were significantly decreased at 1, 4, 8 and 10 weeks of diabetes relative to the control group (p < 0.001). Ca(2+)-ATPase activity was 0.084 +/- 0.008 U l(-1), 0.029 +/- 0.005 U l(-1), 0.029 +/- 0.006 U l(-1), 0.033 +/- 0.003 U l(-1) and 0.058 +/- 0.006 U l(-1) (mean +/- SEM) at control, 1, 4, 8 and 10 week of diabetes respectively. The change in calcium levels in diabetic rat brain at 8 and 10 weeks of diabetes was significantly higher than that of the control group (p < 0.05). On the other hand lipid peroxidation measured as TBARS (thiobarbituric acid reactive substances) was significantly higher at 8 and 10 weeks of diabetes (p < 0.05). The increase in lipid peroxidation observed in diabetic rat brain may be partly responsible for the decrease in calcium ATPase activity. 相似文献
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K S Prasada Rao C S Chetty C H Trottman J E Uzodinma D Desaiah 《Cell biochemistry and function》1985,3(4):267-272
Effect of tricyclohexylhydroxytin (plictran) on Ca2+-ATPase activity was studied in rat brain synaptosomes under in vitro and in vivo conditions. Plictran inhibited basal Ca2+-ATPase activity with an IC50 value of 6 nM suggesting its interaction with calcium transport phenomenon. Plictran inhibited calmodulin (CaM) activated Ca2+-ATPase in a concentration-dependent manner. A complete reversal of calmodulin activation of Ca2+-ATPase was observed with 2-3 nM plictran. A 50 per cent decrease of CaM activated Ca2+-ATPase was observed with 0.5 nM plictran, a concentration at which no significant effect was observed on basal enzyme activity. Of all the brain fractions studied, calmodulin levels in P2 fractions alone were reduced significantly to about 75 per cent of control values in plictran treated rats. The synaptosomal Ca2+-ATPase was also decreased by 35 per cent, 42 per cent and 65 per cent in 10, 20 and 40 mg plictran kg-1 day-1 treated rats for 3 days respectively. The activity levels of Ca2+-ATPase in 10 and 20 mg plictran kg-1 day-1 treated rats were restored to normal level by exogenously added calmodulin. These results suggest that plictran may disrupt synaptic function by altering calcium and calmodulin regulated processes in the central nervous system. 相似文献
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钙池排空操纵的外钙内流决定甘草诱导MGC-803细胞凋亡 总被引:4,自引:0,他引:4
用 EGTA螯合胞外 Ca2 +和异搏定抑制钙通道 ,研究胞外 Ca2 +在甘草诱导 MGC- 80 3细胞中的作用 .流式细胞仪检测凋亡峰和 DNA ladder分析均表明 ,EGTA和异博定阻断细胞凋亡 .分别以 PI或 Rh1 2 3活染后的相应荧光强度表示细胞膜通透性和线粒性膜电位 (ΔΨm) .结果表明 ,细胞膜通透性增强和线粒体 ΔΨm 下降均为细胞凋亡的早期事件 ,EGTA和异博定均可抑制细胞膜通透性增强 ,但 EGTA促进线粒体 ΔΨm 下降 ,而异博定作用相反 .进一步经 PI和 Hoechst33342荧光双染后同时观察细胞膜通透性和细胞核形态 .结果表明 ,凋亡细胞均可 PI着色 ,EGTA和异博定完全阻断染色质凝聚 ,但不能完全抑制细胞膜通透性变化 .借助 Ca2 +探针 Fluo- 3/AM研究凋亡时胞内游离钙的时相变化 ,发现 Ca2 +升高也是细胞凋亡的早期事件 . EGTA和异博定轻微促进凋亡早期 Ca2 +升高 ,但抑制随后 Ca2 +的继续升高 .所有结果提示 ,钙池排空操纵的外 Ca2 +内流在甘草诱导 MGC- 80 3细胞凋亡中发挥决定性的作用 . 相似文献
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G Nicaise K Maggio S Thirion M Horoyan E Keicher 《Biology of the cell / under the auspices of the European Cell Biology Organization》1992,75(2):89-99
The review focuses on calcium accumulation by secretory organelles. The observation that secretory granules contain variable and often important quantities of calcium (1-200 mM of total calcium) can be interpreted as a maturation index. A progressive loading with calcium would be permitted by a Ca2(+)-transport mechanism on the granular membrane and calcium-binding molecules in the granular core. The saturation of this store by the stimulus-induced calcium transient would permit in mature (calcium-loaded) granules the ionic crisis leading to exocytosis. The inside of secretory organelles being acidic, calcium influx into the granule can be driven by calcium-proton exchange. The calcium-proton exchanger could be a Ca2(+)-ATPase. 相似文献