Cd2+处理对5个柳树无性系气体交换参数及叶绿素荧光参数的影响

该实验以筛选的速生、抗逆性强的5个柳树无性系(‘竹柳’、‘苏柳172’、‘苏柳795’、‘旱快柳’、‘旱柳’)一年生扦插幼苗为试验材料,设置对照、轻度(30mg·L-1)、中度(60mg·L-1)和重度(90mg·L-1)4个重金属Cd2+处理水平,通过水培方式探究Cd2+对各柳树无性系气体交换参数、荧光参数及叶绿素含量的影响。结果显示:(1)在Cd2+浓度为30mg·L-1时,导致‘苏柳172’、‘苏柳795’和‘旱快柳’净光合速率降低的原因主要是气孔因素;当Cd2+浓度≥60mg·L-1时,‘苏柳172’、‘苏柳795’、‘旱快柳’及‘竹柳’净光合速率下降的主要原因是非气孔因素,而‘旱柳’在整个Cd2+胁迫过程中净光合速率下降的因素均属于非气孔限制型。(2)整个Cd2+胁迫过程中,‘苏柳172’、‘苏柳795’、‘旱快柳’和‘旱柳’叶片叶绿素含量和Chl a/Chl b值与对照相比均呈下降趋势,而竹柳先上升后下降至低于对照水平。(3)随着Cd2+胁迫程度的增加,叶片PSⅡ最大光化学量子产量(Fv/Fm)和光化学淬灭(qP)整体呈下降趋势,非光化学淬灭系数(NPQ)呈逐渐上升的趋势,而表观光合电子传递速率(ETR)先增加后下降,但5个柳树无性系叶绿素荧光参数的变化幅度以‘竹柳’最小,‘旱柳’最大。研究表明,5个柳树无性系均能在Cd2+浓度为30mg·L-1的培养液中正常生长,且竹柳的光合作用较对照有略微增加,且在90mg·L-1 Cd2+浓度下各无性系生长虽受到抑制但仍能存活,说明它们对重金属Cd2+胁迫都有一定的耐受性,并以‘竹柳’的耐受性最强。     

盐旱交叉胁迫对柽柳幼苗渗透调节物质含量的影响

以2年生柽柳幼苗为试验材料,采用盆栽模拟试验研究不同盐分和干旱胁迫对其叶片中渗透调节物质的影响,以探讨柽柳幼苗对盐旱交叉胁迫的适应性.结果表明:(1)随盐旱胁迫的不断加剧,幼苗叶片中可溶性糖含量呈先升高后降低的趋势,且中度和重度盐旱胁迫下均显著高于对照(CK).(2)幼苗叶片中脯氨酸含量在不同盐旱胁迫下均呈逐渐上升趋势,但在重度盐分和中度、重度干旱交叉胁迫下显著高于CK.(3)幼苗叶片中Na+、Cl含量在不同干旱胁迫下,随盐胁迫的加剧呈不同的变化规律,盐旱胁迫的各个处理水平下均显著高于CK,而K+、Ca2+、SO42-含量在轻度和重度干旱胁迫下随盐胁迫增强不断降低.(4)在中度盐旱胁迫下,K+、Ca2+含量与CK无明显差异.研究表明,柽柳幼苗中渗透调节物质在其抗旱耐盐性方面具有积极的调节作用;柽柳幼苗在盐旱胁迫下表现出一定的交叉适应性,适度的干旱胁迫能增强柽柳幼苗对盐分胁迫的耐受能力.     

不同瑞典能源柳无性系对干旱胁迫的生理响应

以瑞典能源柳无性系2、4、C、E的一年生盆栽扦插苗为材料,采用土壤持续强化干旱胁迫实验,系统测定不同胁迫阶段叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性,以及丙二醛(MDA)、可溶性蛋白含量和株高净生长量,并采用隶属函数模型综合评定了不同能源柳无性系的耐旱性。结果表明:(1)各能源柳无性系叶片的保护酶活性随干旱胁迫的加剧均呈现先增后降趋势:土壤水分含量>11.4%时,SOD、POD和CAT活性均呈增长趋势,三者协同作用;土壤水分含量<11.4%,能源柳无性系2、4的CAT和POD活性显著降低,细胞膜受损。(2)各无性系MDA含量随干旱胁迫的加剧呈增长趋势,土壤含水量降至11.4%时,MDA含量增长显著,其中能源柳无性系4变化最为剧烈。(3)各无性系可溶性蛋白含量随干旱胁迫的持续而增加,但当土壤含水量低于8.1%时有所下降。(4)各能源柳无性系胁迫阶段株高净生长量随干旱胁迫的加剧而降低。(5)隶属函数综合分析发现,4个无性系耐旱潜力表现为能源柳C>能源柳E>能源柳2>能源柳4。     

干旱胁迫对枇杷生理特性及生长的影响

以3年生‘大五星’枇杷嫁接苗为试验材料,通过盆栽控水试验设置4个水分处理梯度:对照(CK)、轻度胁迫(LS)、中度胁迫(MS)和重度胁迫(SS),研究不同程度土壤干旱对枇杷幼树的生长和生理特性的影响。结果显示:(1)枇杷的株高、地上和地下生物量随干旱胁迫的增强呈下降趋势。(2)在轻度和中度胁迫下叶片叶绿素含量增加,随着土壤水分的减少,叶绿素含量和叶片相对含水量(LRWC)均显著下降。(3)叶片净光合速率(Pn)、蒸腾速率(Tr)和气孔导度(Gs)随着干旱胁迫的加剧均显著下降,其中Gs下降幅度最大,而胞间CO2浓度(Ci)则表现为先下降后上升,重度胁迫时叶片的水分利用率(WUE)最低。(4)干旱的加重使叶片超氧化物歧化酶(SOD)和过氧化物酶(POD)的活性呈现先增加后降低的趋势,过氧化氢酶(CAT)活性在轻度胁迫下最为活跃但在重度胁迫时显著降低,丙二醛(MDA)含量在中度胁迫第10天开始显著升高。(5)随干旱胁迫的加剧游离脯氨酸(Pro)含量增加且在重度胁迫第10天达到最大值,而可溶性蛋白(SP)含量在胁迫后期与对照无显著差异,可溶性糖(SS)含量在重度胁迫后期达到峰值且与对照差异显著。研究表明,在轻度和中度干旱胁迫下,枇杷叶片光合作用受到抑制,但能够积极调控抗氧化酶的活性和渗透调节物质的含量等来增强耐受性,而重度胁迫下,叶片膜系统和光合系统受到损伤,枇杷生长受到严重抑制。     

5种速生柳无性系对Pb~(2+)胁迫的生理抗性比较

以5种一年生速生柳(竹柳、苏柳172、苏柳795、旱快柳、旱柳)幼苗为材料,通过盆栽铅胁迫实验,测定其生长过程中的生理生化指标,分析其对铅胁迫的生理响应及耐受性。结果表明:(1)随着Pb2+浓度的增加,5种速生柳叶片保护酶SOD、POD和CAT活性均呈先升高后降低的趋势,其中竹柳的3种保护酶活性均为最高。(2)随着Pb2+浓度的增加,各无性系叶片中渗透性调节物质可溶性蛋白、脯氨酸和可溶性糖含量均呈先上升后下降的趋势,竹柳和苏柳172在高Pb2+浓度(1500mg·kg-1)胁迫下表现出对Pb2+的强耐受性。(3)随着Pb2+浓度的增加,各速生柳无性系叶片丙二醛含量均呈逐渐增高的趋势,但增幅有所不同,竹柳增幅最小,旱柳增幅最大。(4)随着Pb2+浓度的增加,5种速生柳叶片中叶绿素含量均呈先上升后下降的趋势,但各无性系的下降幅度存在差异,其中竹柳的下降幅度最小,旱柳下降幅度最大。(5)随着Pb2+浓度的增加,5种速生柳根系活力都呈先上升后下降的趋势,其中竹柳的根系活力最强,苏柳795的最弱。(6)各速生柳根系对铅均有一定的吸附能力,竹柳对铅积累量最大。(7)对各生理指标的Fuzzy数学隶属函数综合评价结果表明,5种速生柳对Pb2+耐受性强弱表现为:竹柳旱快柳旱柳苏柳795苏柳172。     

外源亚精胺对Hg2+胁迫下凤眼莲抗氧化系统的影响

通过Hoagland溶液培养实验,研究了外源亚精胺(Spd)(0.1 mmol?L-1)对Hg2+(0、10、20、30和40μmol?L-1)胁迫下凤眼莲叶片细胞内叶绿素、可溶性糖、可溶性蛋白含量及抗氧化系统的调节作用.结果显示,(1)随Hg2+处理浓度的升高,各处理凤眼莲叶片叶绿素a(Chl a)和叶绿素b(Chl b)含量均先升后降,并均在10μmol?L-1时达到最高值,但外源Spd处理组显著高于相应对照.(2)各处理凤眼莲叶片可溶性蛋白含量随Hg2+处理浓度的升高也呈先升后降趋势,而可溶性糖含量则呈持续上升趋势,但外源Spd处理亦明显高于相应对照.(3)随Hg2+处理浓度的升高,抗氧化物质AsA和GSH含量及抗氧化酶SOD、CAT、POD、APX及GR活性也均呈先升后降的变化趋势,而外源Spd处理植株的含量和活性均显著高于相应对照.(4)各处理凤眼莲叶片的H2O2、MDA含量及O2?-产生速率随Hg2+处理浓度的升高均持续上升,但在外源Spd处理后均比对照组下降.研究表明,Hg2+胁迫使凤眼莲生长受到严重伤害,外源Spd可大幅度地提高其抗氧化物质含量和保护酶活性,从而增强凤眼莲抗Hg2+胁迫的能力.     

不同能源柳无性系对NaCl胁迫的生理响应

为了探索不同能源柳无性系的耐盐潜力,在盆栽条件下,对1年生能源柳无性系2、4、C、E扦插苗设置NaCl盐分梯度,使其土壤含盐量保持在0、0.2%、0.4%、0.6%、0.8%,并以乡土旱柳为对照,胁迫45d后,对5个无性系的气体交换参数、保护酶活性和有机渗透调节物质进行测定。研究发现:(1)在低浓度盐胁迫下(土壤含盐量0 0.4%),导致各能源柳光合速率降低的是气孔因素;在高浓度盐胁迫下(土壤含盐量大于0.4%),导致各能源柳无性系光合速率下降的主要是非气孔因素。(2)随着土壤盐浓度的增加,各能源柳无性系的过氧化氢酶(CAT)活性逐渐减小,过氧化物酶(POD)活性先增加后减小,而超氧化物歧化酶(SOD)活性呈现增加的趋势,并且4个能源柳无性系在盐胁迫条件下保护酶活性强于旱柳。(3)在土壤含盐量小于0.6%时,4个能源柳无性系的可溶性蛋白含量始终大于旱柳;在整个胁迫过程中,能源柳C的脯氨酸含量始终大于旱柳。(4)与旱柳相比,4个能源柳无性系在土壤含盐量0 0.4%的土壤中都能良好生长,其中能源柳C的耐盐潜力更大,可在土壤含盐量为0.6%的土壤中正常生长。这说明,4个能源柳无性系均可在天津轻、中度盐渍化地区栽培。     

大青杨对不同干旱胁迫强度的形态和生理响应

为了研究大青杨对干旱环境的耐受性,通过聚乙二醇(PEG-6000)模拟干旱胁迫,对不同胁迫强度下大青杨的生长状态、生物量分配、生理生化应答特征进行测定分析。结果显示,随着干旱胁迫时间的延长和强度的增加,大青杨幼苗的苗高、地径生长量和生物量积累明显减少,轻度、中度、重度胁迫下苗高生长量分别下降了17.71%、20.29%、45.96%,地径生长量下降了4.78%、13.62%、17.29%,根冠比上升了11.93%、32.61%、64.10%。叶片相对含水量呈下降趋势,相对电导率呈上升趋势,轻度、中度、重度胁迫下相对含水量分别比对照低23.60%、38.56%和66.78%,相对电导率比对照高106.97%、161.84%、241.75%,说明干旱胁迫对叶片造成了明显的损伤。脯氨酸、可溶性蛋白、可溶性糖含量均呈现先上升再下降的趋势,且轻度胁迫比中度、重度胁迫变化幅度小。在胁迫前期和中期均高于对照,说明大青杨幼苗有一定的抗旱能力。在胁迫末期有低于对照的趋势,此时幼苗细胞、组织受到了不可逆转的伤害。大青杨叶片的净光合速率明显呈现双峰模式,受胁迫幼苗显著低于对照且到达峰值的时间不同。胁迫后幼苗的第一个峰值出现的时间比对照提前2h。大青杨能够忍耐短时轻度的干旱胁迫环境,中度和重度干旱胁迫对大青杨幼苗期的生长和生理过程有显著影响。     

干旱胁迫对杠柳光合特性及抗氧化酶活性的影响

以盆栽杠柳幼苗为材料,用称重控水的方法设置4个含水量梯度,研究不同程度干旱胁迫对杠柳光合特性和抗氧化系统的影响,为黄河三角洲贝壳堤岛植被恢复过程中的物种培育和种植管理提供依据.结果表明:(1)随着干旱胁迫的加剧,杠柳幼苗叶片净光合速率(Pn)先增后降,蒸腾速率(Tr)和气孔导度(Gs)逐渐下降,胞间CO2浓度(Ci)先降后升,说明杠柳幼苗Pn下降在轻度干旱胁迫下主要是由气孔导度下降引起,而在重度胁迫下主要由非气孔因素引起.(2)随着干旱胁迫程度的加剧,杠柳幼苗瞬时水分利用效率(WUE)、表观光能利用效率(LUE)、羧化效率(CE)均呈现先增加后下降趋势;WUE在重度胁迫下才开始下降,比对照显著下降了39%,LUE(CE)则在中度胁迫下就开始下降,中度和重度胁迫分别比对照下降了48%(33%)、71%(69%).(3)随着干旱胁迫的加剧,杠柳叶片的超氧化物歧化酶(SOD)活性先升高后降低,过氧化物酶(POD)、过氧化氢酶(CAT)活性和MDA含量均逐渐增高,表明POD、CAT抑制杠柳叶片脂质过氧化效应不显著,SOD则抑制作用显著,且12.8%的贝壳沙含水量是SOD的耐受阈值,此时MDA含量比对照显著增加153%.     

盐碱胁迫对欧李叶片部分生理生化指标的影响

以盆栽欧李为试验材料,研究了轻度、中度和重度盐碱胁迫下其叶片部分生理生化指标的变化,探讨欧李植株的耐盐碱能力.结果表明:(1)欧李叶片的MDA含量在轻度和中度盐碱胁迫处理下与对照变化趋势相似,而在重度胁迫下从开始即以较快的速度增加;其叶片细胞膜透性在中度和重度胁迫下明显增大,而轻度胁迫对此基本没有影响.(2)欧李叶片可溶性糖含量随盐碱胁迫处理而明显改变,叶片脯氨酸含量随着盐碱胁迫程度的增强和处理时间的延长而增加;叶片的束缚水/自由水在轻度和中度胁迫处理下较大,而重度胁迫处理下则较小.(3)欧李叶片的叶绿素含量在各盐碱处理末期均显著下降,但其在处理的不同阶段反应不同;类胡萝卜素含量受盐碱胁迫影响较大,其在长期盐碱胁迫下显著下降,但其在胁迫较轻或胁迫时间较短条件下反而增加.研究发现,欧李植株可通过调节叶片可溶性糖和脯氨酸含量以及束缚水/自由水比值来提高其耐盐碱能力.     

Palytoxin Acts on Na+,K+-ATPase but not Nongastric H+,K+-ATPase

Palytoxin (PTX) opens a pathway for ions to pass through Na,K-ATPase. We investigate here whether PTX also acts on nongastric H,K-ATPases. The following combinations of cRNA were expressed in Xenopus laevis oocytes: Bufo marinus bladder H,K-ATPase α₂- and Na,K-ATPase β₂-subunits; Bufo Na,K-ATPase α₁- and Na,K-ATPase β₂-subunits; and Bufo Na,K-ATPase β₂-subunit alone. The response to PTX was measured after blocking endogenous Xenopus Na,K-ATPase with 10 μm ouabain. Functional expression was confirmed by measuring ⁸⁶Rb uptake. PTX (5 nm) produced a large increase of membrane conductance in oocytes expressing Bufo Na,K-ATPase, but no significant increase occurred in oocytes expressing Bufo H,K-ATPase or in those injected with Bufo β₂-subunit alone. Expression of the following combinations of cDNA was investigated in HeLa cells: rat colonic H,K-ATPase α₁-subunit and Na,K-ATPase β₁-subunit; rat Na,K-ATPase α₂-subunit and Na,K-ATPase β₂-subunit; and rat Na,K-ATPase β₁- or Na,K-ATPase β₂-subunit alone. Measurement of increases in ⁸⁶Rb uptake confirmed that both rat Na,K and H,K pumps were functional in HeLa cells expressing rat colonic HKα₁/NKβ₁ and NKα₂/NKβ₂. Whole-cell patch-clamp measurements in HeLa cells expressing rat colonic HKα₁/NKβ₁ exposed to 100 nm PTX showed no significant increase of membrane current, and there was no membrane conductance increase in HeLa cells transfected with rat NKβ₁- or rat NKβ₂-subunit alone. However, in HeLa cells expressing rat NKα₂/NKβ₂, outward current was observed after pump activation by 20 mm K⁺ and a large membrane conductance increase occurred after 100 nm PTX. We conclude that nongastric H,K-ATPases are not sensitive to PTX when expressed in these cells, whereas PTX does act on Na,K-ATPase.     

The sodium cycle: A novel type of bacterial energetics

The progress of bioenergetic studies on the role of Na⁺ in bacteria is reviewed. Experiments performed over the past decade on several bacterial species of quite different taxonomic positions show that Na⁺ can, under certain conditions, substitute for H⁺ as the coupling ion. Various primary Na⁺ pumps ( generators) are described, i.e., Na⁺-motive decarboxylases, NADH-quinone reductase, terminal oxidase, and ATPase. The formed is shown to be consumed by Na⁺ driven ATP-synthase, Na⁺ flagellar motor, numerous Na⁺, solute symporters, and the methanogenesis-linked reverse electron transfer system. InVibrio alginolyticus, it was found that , generated by NADH-quinone reductase, can be utilized to support all three types of membrane-linked work, i.e., chemical (ATP synthesis), osmotic (Na⁺, solute symports), and mechanical (rotation of the flagellum). InPropionigenum modestum, circulation of Na⁺ proved to be the only mechanism of energy coupling. In other species studied, the Na⁺ cycle seems to coexist with the H⁺ cycle. For instance, inV. alginolyticus the initial and terminal steps of the respiratory chain are Na⁺ - and H⁺-motive, respectively, whereas ATP hydrolysis is competent in the uphill transfer of Na⁺ as well as of H⁺. In the alkalo- and halotolerantBacillus FTU, there are H⁺ - and Na⁺-motive terminal oxidases. Sometimes, the Na⁺-translocating enzyme strongly differs from its H⁺-translocating homolog. So, the Na⁺-motive and H⁺-motive NADH-quinone reductases are composed of different subunits and prosthetic groups. The H⁺-motive and Na⁺-motive terminal oxidases differ in that the former is ofaa ₃-type and sensitive to micromolar cyanide whereas the latter is of another type and sensitive to millimolar cyanide. At the same time, both Na⁺ and H⁺ can be translocated by one and the sameP. modestum ATPase which is of the F₀F₁-type and sensitive to DCCD. The sodium cycle, i.e., a system composed of primary generator(s) and consumer(s), is already described in many species of marine aerobic and anaerobic eubacteria and archaebacteria belonging to the following genera:Vibrio, Bacillus, Alcaligenes, Alteromonas, Salmonella, Klebsiella, Propionigenum, Clostridium, Veilonella, Acidaminococcus, Streptococcus, Peptococcus, Exiguobacterium, Fusobacterium, Methanobacterium, Methanococcus, Methanosarcin, etc. Thus, the sodium world seems to occupy a rather extensive area in the biosphere.     

Role of the furosemide-sensitive Na+/K+ transport system in determining the steady-state Na+ and K+ content and volume of human erythrocytesin vitro andin vivo

Summary To study the physiological role of the bidirectionally operating, furosemide-sensitive Na⁺/K⁺ transport system of human erythrocytes, the effect of furosemide on red cell cation and hemoglobin content was determined in cells incubated for 24 hr with ouabain in 145mm NaCl media containing 0 to 10mm K⁺ or Rb⁺. In pure Na⁺ media, furosemide accelerated cell Na⁺ gain and retarded cellular K⁺ loss. External K⁺ (5mm) had an effect similar to furosemide and markedly reduced the action of the drug on cellular cation content. External Rb⁺ accelerated the Na⁺ gain like K⁺, but did not affect the K⁺ retention induced by furosemide. The data are interpreted to indicate that the furosemide-sensitive Na⁺/K⁺ transport system of human erythrocytes mediates an equimolar extrusion of Na⁺ and K⁺ in Na⁺ media (Na⁺/K⁺ cotransport), a 1:1 K⁺/K⁺ (K⁺/Rb⁺) and Na⁺/Na⁺ exchange progressively appearing upon increasing external K⁺ (Rb⁺) concentrations to 5mm. The effect of furosemide (or external K⁺/Rb⁺) on cation contents was associated with a prevention of the cell shrinkage seen in pure Na⁺ media, or with a cell swelling, indicating that the furosemide-sensitive Na⁺/K⁺ transport system is involved in the control of cell volume of human erythrocytes. The action of furosemide on cellular volume and cation content tended to disappear at 5mm external K⁺ or Rb⁺. Thein vivo red cell K⁺ content was negatively correlated to the rate of furosemide-sensitive K⁺ (Rb⁺) uptake, and a positive correlation was seen between mean cellular hemoglobin content and furosemide-sensitive transport activity. The transport system possibly functions as a K⁺ and waterextruding mechanism under physiological conditiosin vivo. The red cell Na⁺ content showed no correlation to the activity of the furosemide-sensitive transport system.     

Sodium ATPase and Sodium/proton Antiporter Are Not Obligatory for Sodium Homeostasis of Enterococcus hirae at Acid pH

Enterococcus hirae grows in a broad pH range from 5 to 11. An E. hirae mutant 7683 lacking the activities of two sodium pumps, Na⁺-ATPase and Na⁺/H⁺ antiporter, does not grow in high Na⁺ medium at pH above 7.5. We found that 7683 grew normally in high Na⁺ medium at pH 5.5. Although an energy-dependent sodium extrusion at pH 5.5 was missing, the intracellular levels of Na⁺ and K⁺ were normal in this mutant. The Na⁺ influx rates of 7683 and two other strains at pH 5.5 were much slower than those at pH 7.5. These results suggest that Na⁺ elimination of this bacterium at acid pH is achieved by a decrease in Na⁺ entry and a normal K⁺ uptake.     

Nigericin-induced Na+/H+ and K+/H+ exchange in synaptosomes: Effect on [3H]GABA release

The effect of the putative K⁺/H⁺ ionophore, nigericin on the internal Na⁺ concentration ([Na _i ]), the internal pH (pH _i ), the internal Ca²⁺ concentration ([Ca _i ]) and the baseline release of the neurotransmitter, GABA was investigated in Na⁺-binding benzofuran isophtalate acetoxymethyl ester (SBFIAM), 2′,7′-bis(carboxyethyl)-5(6) carboxyfluorescein acetoxymethyl ester (BCECF-AM), fura-2 and [³H]GABA loaded synaptosomes, respectively. In the presence of Na⁺ at a physiological concentration (147 mM), nigericin (0.5 μM) elevates [Na _i ] from 20 to 50 mM, increases thepH _i , 0.16 pH units, elevates four fold the [Ca _i ] at expense of external Ca²⁺ and markedly increases (more than five fold) the release of [³H]GABA. In the absence of a Na⁺ concentration gradient (i.e. when the external Na⁺ concentration equals the [Na _i ]), the same concentration (0.5 μM) of nigericin causes the opposite effect on thepH _i (acidifies the synaptosomal interior), does not modify the [Na _i ] and is practically unable to elevate the [Ca _i ] or to increase [³H]GABA release. Only with higher concentrations of nigericin than 0.5 μM the ionophore is able to elevate the [Ca _i ] and to increase the release of [³H]GABA under the conditions in which the net Na⁺ movements are eliminated. These results clearly show that under physiological conditions (147 mM external Na⁺) nigericin behaves as a Na⁺/H⁺ ionophore, and all its effects are triggered by the entrance of Na⁺ in exchange for H⁺ through the ionophore itself. Nigericin behaves as a K⁺/H⁺ ionophore in synaptosomes just when the net Na⁺ movements are eliminated (i.e. under conditions in which the external and the internal Na⁺ concentrations are equal). In summary care must be taken when using the putative K⁺/H⁺ ionophore nigericin as an experimental tool in synaptosomes, as under standard conditions (i.e. in the presence of high external Na⁺) nigericin behaves as a Na⁺/H⁺ ionophore.     

Intracellular calcium content of human erythrocytes: Relation to sodium transport systems

Summary To study the possible role of intracellular Ca (Ca _i ) in controlling the activities of the Na⁺–K⁺ pump, the Na⁺–K⁺ cotransport and the Na⁺/Li⁺ exchange system of human erythrocytes, a method was developed to measure the amount of Ca embodied within the red cell. For complete removal of Ca associated with the outer aspect of the membrane, it proved to be essential to wash the cells in buffers containing less than 20nm Ca. Ca was extracted by HClO₄ in Teflon^® vessels boiled in acid to avoid Ca contaminations and quantitated by flameless atomic absorption. Ca _i of fresh human erythrocytes of apparently healthy donors ranged between 0.9 and 2.8 mol/liter cells. The mean value found in females was significantly higher than in males. The interindividual different Ca contents remained constant over periods of more than one year. Sixty to 90% of Ca _i could be removed by incubation of the cells with A23187 and EGTA. The activities of the Na⁺–K⁺ pump, of Na⁺–K⁺ cotransport and Na⁺/Li⁺ exchange and the mean cellular hemoglobin content fell with rising Ca _i ; the red cell Na⁺ and K⁺ contents rose with Ca _i . Ca depletion by A23187 plus EGTA as well as chelation of intracellular Ca²⁺ by quin-2 did not significantly enhance the transport rates. It is concluded that the large scatter of the values of Ca _i of normal human erythrocytes reported in the literature mainly results from a widely differing removal of Ca associated with the outer aspect of the membrane.     

Functional Consequences of Various Leucine Mutations in the M3/M4 Loop of the Na<Superscript>+</Superscript>,K<Superscript>+</Superscript>-ATPase α-Subunit

Leucines were mutated within the sequence L³¹¹ILGYTWLE³¹⁹ of the extracellular loop flanking the third (M3) and fourth (M4) transmembrane segments (M3/M4 loop) of the Torpedo Na⁺,K⁺-ATPase α-subunit. Replacement of Leu³¹¹ with Glu resulted in a considerable loss of Na⁺,K⁺-ATPase activity. Replacement of Leu³¹³ with Glu shifted the equilibrium of E₁P and E₂P toward E₁P and reduced the rate of the E₁P to E₂P transition. The reduction of the transition rate and stronger inhibition of Na⁺,K⁺-ATPase activity by Na⁺ at higher concentrations together suggest that there is interference of Na⁺ release on the extracellular side in the Leu³¹³ mutant. Thus, Leu³¹³ could be in the pathway of Na⁺ exit. Replacement of Leu³¹⁸ with Glu yielded an enzyme with significantly reduced apparent affinity for both vanadate and K⁺, with an equilibrium shifted toward E₂P and no alteration in the transition rate. The reduced vanadate affinity is due to the lower rate of production of vanadate-reactive [K⁺ ₂]E₂ caused by inhibition of dephosphorylation through reduction of the K⁺ affinity of E₂P. Thus, Leu³¹⁸ may be a critical position in guiding external K⁺ to its binding site.     

蛋白质感染颗粒与二价铜离子

蛋白质感染颗粒（PrP）的错误折叠被认为是引起一些神经退化性疾病的主因,但其正常构象（PrP^C）的功能却一直不为人所知．近年来研究发现,在正常细胞中,尤其是脑细胞中,细胞膜PrP^C可通过内吞作用进入细胞质而将Cu²⁺载运至SOD1,从而参与调节SOD1 的活性及细胞铜代谢．另有研究表明,Cu²⁺对于PrP^Sc（错误构象）的蛋白水解酶K抗性的恢复及不同“病株”的形成也有很重要的作用.