土壤盐碱胁迫对春小麦K^＋、Na^＋选择性吸收的影响

通过对2种浓度土壤盐分胁迫下春小麦各品种不同时期各器官K^ 、Na^ 含量以及K^ /Na^ 的变化及其与抗盐性的关系研究,结果表明,随土壤盐浓度的升高,各品种的产量及各农艺性状值均有所下降,但不同品种的下降程度不同。随土壤盐浓度的升高,植株中K^ 、Na^ 含量均有所增加,但K^ 增加的幅度小于Na^ 的增加幅度,因而K^ /Na^ 呈明显下降趋势;在不同土壤盐分胁迫下,小麦品种K^ 、Na^ 随生育进程在体内各器官的分配发生动态变化,在分蘖期地上部K^ /Na^ ＞根部,孕穗期各器官K^ /Na^ 依次为：幼穗＞旗叶＞茎＞倒4叶,而灌浆期则依次为：籽粒＞旗叶＞茎＞倒4叶,说明生长旺盛的器官拒Na^ 能力强于其它器官;不同品种的K^ 、Na^ 含量及K^ /Na^ 不同,一般抗盐性强的品种在各时期均具有较高的K^ /Na^ ,反之则K^ /Na^ 较低;小麦的籽粒产量在一定范围内与其植株地上部各器官的K^ /Na^ 中一定的正相关,其中与分蘖期植株地上部的K^ /Na^ 及叶（K^ /Na^ ）／根（K^ /Na^ ）呈极显著正相关,而与此时期的SNa^ K^ 相关性最强,γ为－0．9670。因而,以分蘖期的K^ /Na^ 尤其是SNa^ K^ 作为小麦田间抗盐性的指标,具有一定的可靠性。     

珠江口池养梭鱼鳃的光镜、扫描和透射电镜观察

应用光学显微镜、扫描电镜和透射电镜对珠江口池塘养殖梭鱼Liza haematocheila鳃的组织结构、表面形态特征及鳃小片超微结构进行了观察。结果表明梭鱼具有4对鳃,每个鳃由鳃弓、鳃丝、鳃小片和鳃耙组成。梭鱼鳃丝和鳃小片的表面结构和超微结构与其他硬骨鱼类的基本相似,鳃丝表面分布有众多规则或不规则的环形微嵴、沟、坑、孔等结构。鳃丝分为呼吸区和非呼吸区,呼吸区较为平滑,上皮细胞表面无微嵴,呈皱褶状;非呼吸区分布有沟、坑、孔等结构,上皮细胞有较规则的指纹状微嵴。鳃小片是最主要的呼吸场所,由基膜、上皮细胞、内皮细胞、柱细胞和毛细血管网组成。泌氯细胞主要分布在鳃小片基部,并有开口通往外界。本文还探讨了梭鱼鳃的结构与其功能的密切关系。     

He—Ne激光辐照与UV-B辐射对小麦幼苗细胞器中Na^＋／K^＋-ATP酶活性的影响

分别采用5mJ·s^-1·mm^-2He-Ne激光辐照、10．08kJ·m·^-2,d^-1增强UV-B辐射及二者组合对小麦（Triticum aestivum）晋麦8号（Triticum aestivum‘Jinmai8’）幼苗进行处理。第5天开始测定各处理小麦幼苗叶片中线粒体、叶绿体及细胞溶质中Na^＋／K^＋-ATP酶活性的变化。结果表明,随着处理天数的增加,小麦幼苗叶片线粒体、叶绿体及细胞溶质中Na^＋／K^＋-ATP酶活性均在第6天下降,第7天升高,而后又逐渐下降。在处理的第7天,仅He—Ne激光辐照可使小麦幼苗叶片线粒体、叶绿体及细胞溶质中Na^＋／K^＋-ATP酶活性升高：增强UV-B辐射使各细胞器中Na^＋／K^＋-ATP酶活性下降：复合处理后小麦各细胞器中Na^＋／K^＋-ATP酶活性均高于UV-B单独辐射处理。实验结果表明,一定剂量的He-Ne激光辐照能够部分修复UV-B辐射对小麦幼苗细胞器中Na^＋／K^＋-ATP酶造成的损伤。     

菲对斑马鱼鳃和肝组织结构的影响

采用浓度为0、0.05和100.0 μg·L-1的菲水溶液对斑马鱼(Brachydanio rerio)进行36 d的暴露实验,研究了菲对斑马鱼鳃、肝结构的影响.H-E染色显示:在0.05和100.0 μg·L-1的菲溶液暴露下,受试鱼的鳃受到损伤,均发生鳃小片上皮细胞肥大和水肿;菲浓度为100.0 μg·L-1时,受试鱼还发生鳃丝上皮增厚、鳃小片上皮隆起现象.此外,菲暴露造成斑马鱼的肝组织损伤,0.05 μg·L-1的菲溶液暴露下,受试鱼的肝细胞发生肿大,胞质产生空泡;菲浓度为100.0 μg·L-1时,受试鱼的肝细胞变得不规则,细胞核萎缩变形和偏离细胞中心,部分肝细胞空泡化程度加重,发生核溶解或细胞溶解,造成局部肝组织坏死.表明水环境中菲浓度达到0.05 μg·L-1即已对斑马鱼的鳃、肝产生毒性作用;而菲浓度达到100.0 μg·L-1时,受试鱼的鳃、肝将受到较严重的损伤;伴随菲浓度的升高,鱼的鳃、肝损伤加重.     

盐度对大麻哈鱼幼鱼存活率、鳃ATP酶活力及其组织结构的影响

为研究不同盐度对大麻哈鱼幼鱼存活率、鳃ATP酶活力及其组织结构的影响, 试验共设置4个盐度组(S0、S8、S16、S24), 试验周期42d, 解剖取鱼鳃测定ATP酶活力, 并运用组织切片及扫描电镜技术观察其鳃组织结构的变化。结果显示: S8和S16组大麻哈鱼幼鱼存活率最高, 均达到98.89%, S0组存活率为94.45%, 而S24组存活率最低, 为83.34%。随着盐度的升高, 大麻哈鱼幼鱼鳃组织Na+/K+-ATP和Ca2+/Mg2+-ATP酶活力均呈现出先升高后下降的趋势且酶活力最高的均为S8组、最低的为S24组。鳃丝宽度随盐度升高逐渐增大且各组之间呈现出显著性差异(P<0.05), 而鳃小片长度和宽度均随盐度升高逐渐减小。扫描电镜结果表明随着盐度的升高线粒体丰富细胞数量逐渐增多、顶膜变小且微绒毛消失; 同S0组相比, S8组和S16组鳃丝表面扁平上皮细胞之间的轮廓更加清楚且环形微脊条纹清晰, 而S24组鳃丝表面扁平上皮细胞之间界限模糊, 环形微脊间有融合或间断情况; 鳃小片底部扁平上皮细胞表层结构由清晰逐渐变得混乱、气孔数量逐渐减少且孔径变小。因此推测大麻哈鱼幼鱼在降海阶段适宜的盐度生存范围可能介于8‰—16‰, 具体有待进一步研究。     

久效磷对中国对虾细胞超微结构的影响Ⅲ.对鳃的毒性效应

对中国对虾鳃细胞超微结构的透射电镜观察结果表明 ,经 0 .1 0mg·L- 1 久效磷处理的中国对虾 ,鳃丝细胞的内质网水肿、扩张和囊泡化 ,粗面内质网膜上的核糖体颗粒大量脱落 ;高尔基膜囊轻微水肿 ,高尔基小泡水肿扩张 ,严重者破裂 ;线粒体的内嵴膨胀 ,局部瓦解乃至解体 ;鳃丝上皮细胞角质层变性脱落 ,核膜水肿 ,部分溶解 .     

慢性氨氮胁迫对台湾泥鳅幼鱼生长、免疫及组织结构的影响

以平均体质量为(2.00±0.56) g的台湾泥鳅(Paramisgurnus dabryanus ssp. Taiwan)幼鱼为研究对象, 探究氨氮胁迫浓度为0、3.5、6.0、10.4和18.0 mg/L时台湾泥鳅幼鱼生长性能、免疫酶活性及组织结构的变化。经过56d的氨氮胁迫试验结果显示: 台湾泥鳅终末体质量(W_F)、增重率(WGR)和特定生长率(SGR)随氨氮浓度的升高而逐渐降低, 各组之间存活率无显著差异。10.4和18.0 mg/L处理组肝脏超氧化物歧化酶(SOD)、碱性磷酸酶(ALP)和鳃Na+K+-ATP酶活性均显著低于对照组(P<0.05); 18.0 mg/L处理组肝脏溶菌酶(LZM)活性显著低于对照组(P<0.05), 而丙二醛(MDA)含量显著高于对照组(P<0.05)。组织切片结果显示, 氨氮处理组幼鱼均存在不同程度的鳃组织损伤, 表现为鳃小片短小、上皮细胞水肿、细胞空泡化、上皮细胞坏死和脱落; 肝组织则表现为肝血窦扩张、细胞轮廓模糊、细胞水样变性、细胞核偏移和细胞核溶解; 且两种组织中其病变区域随着氨氮浓度升高而增大。上述结果表明, 氨氮胁迫对台湾泥鳅幼鱼的生长和免疫酶活性具有显著的抑制作用, 并对其鳃和肝组织造成损伤。     

番茄碱对人红细胞膜Na+-K+-ATPase活性影响的研究

以低渗法从新鲜健康人红细胞中制得膜Na^ -K^ -ATPase,研究了番茄碱(tomatine)X~人红细胞膜Na^ -K^ -ATPase活性的影响。实验结果表明,反应体系中的tomatine浓度低于1mmol／L时,对不依赖钙调蛋白(CAM)激活的Na^ -K^ -ATPase(称之为基本酶活)影响不大,浓度达1mmol／L时,该酶活性仍保持在95％左右;而在此浓度范围内,tomatine对依赖CaM的Na^ -K^ -ATPase有明显的抑制作用,其IC50值为0．16mmol／L．说明tomatine对膜酶Na^ -K^ -ATPase活性的影响可能是通过阻断CaM激活的途径而起作用,从而为进一步研究番茄碱的作用机制奠定了基础。     

镉对黄颡鱼鳃线粒体结构和能量代谢的影响

采用室内模拟方法,研究了重金属镉对黄颡鱼[Pelteobagrus fulvidraco (Richardson)]鳃线粒体结构和能量代谢的影响及其作用机理.结果表明,50 μg·L^-1组黄颡鱼鳃线粒体结构未受损,且各项测定指标与对照组无显著差异(P＞0.05);500 μg·L^-1组黄颡鱼鳃线粒体严重受损,除LD、MDA、ADP、AMP和血浆K⁺含量显著高于对照组外,其余检测指标均显著低于对照组(P＜0.05).高浓度镉短期暴露将降低鱼鳃线粒体SOD活性而导致线粒体氧化损伤,同时抑制PFK和ATP酶活性, 影响鳃的能量供应和利用,最终降低鳃血浆渗透压和离子浓度调节能力可能是其毒性机理之一.     

新合成复合絮凝剂HECES絮凝性能研究

以元机铝盐和天然高分子玉米淀粉为原料,合成一种生态安全型复合高效絮凝剂HECES．结果表明,在一定范围里适当提高淀粉／铝盐比例,有利于HECES絮凝性能的提高;处理模拟废水时,3．0mg·L^-1的HECES相当于4．5mg·L^-2PAC和1．0mg·L^-1PAM复合投加效果;对生活污水、市政污水的最佳投加量分别为8．0mg·L^-1和4．0mg·L^-1,相当于PAC用量的50％和40％,浊度去除率高达95％和99％．研究还证实絮凝效果与模拟浊度废水的浓度、供试污水的组成成分有关,HECES对高浓度废水处理效果更佳．HECES在PH＝4．0-9.5范围内絮凝效果最佳。其生态安全特点体现在投加量少,具有较大生态毒性的游离态铝离子残留量低。     

Failure to up-regulate gill Na+,K+-ATPase alpha-subunit isoform alpha1b may limit seawater tolerance of land-locked Arctic char (Salvelinus alpinus)

Many populations of Arctic char (Salvelinus alpinus) are land-locked, physically separated from the ocean by natural barriers and unable to migrate to sea like anadromous populations. Previous studies which experimentally transferred land-locked Arctic char to seawater report high mortality rates due to osmoregulatory failure and an inability to up-regulate gill Na(+),K(+)-ATPase activity. This study examined the mRNA expression of two recently discovered alpha-subunit isoforms of gill Na(+)K(+)-ATPase (alpha1a and alpha1b) during seawater exposure of land-locked Arctic char. mRNA levels of these gill Na(+),K(+)-ATPasealpha-subunit isoforms were compared to Na(+),K(+)-ATPase activity and protein levels and related to osmoregulatory performance. Land-locked Arctic char were unable to regulate plasma osmolality following seawater exposure. Seawater exposure did not induce an increase in gill Na(+),K(+)-ATPase activity or protein levels. Na(+),K(+)-ATPase isoform alpha1a mRNA quickly decreased upon exposure to seawater, while isoform alpha1b levels were unchanged. These results suggest the inability of land-locked Arctic char to acclimate to seawater is due a failure to up-regulate gill Na(+),K(+)-ATPase activity which may be due to their inability to increase Na(+),K(+)-ATPase alpha1b mRNA expression.     

K+ and NH4(+) modulate gill (Na+, K+)-ATPase activity in the blue crab, Callinectes ornatus: fine tuning of ammonia excretion

To better comprehend the mechanisms of ionic regulation, we investigate the modulation by Na+, K+, NH4(+) and ATP of the (Na+, K+)-ATPase in a microsomal fraction from Callinectes ornatus gills. ATP hydrolysis obeyed Michaelis-Menten kinetics with KM=0.61+/-0.03 mmol L(-1) and maximal rate of V=116.3+/-5.4 U mg(-1). Stimulation by Na+ (V=110.6+/-6.1 U mg(-1); K0.5=6.3+/-0.2 mmol L(-1)), Mg2+ (V=111.0+/-4.7 U mg(-1); K0.5=0.53+/-0.03 mmol L(-1)), NH4(+) (V=173.3+/-6.9 U mg(-1); K0.5=5.4+/-0.2 mmol L(-1)) and K+ (V=116.0+/-4.9 U mg(-1); K0.5=1.5+/-0.1 mmol L(-1)) followed a single saturation curve, although revealing site-site interactions. In the absence of NH4(+), ouabain (K(I)=74.5+/-1.2 micromol L(-1)) and orthovanadate inhibited ATPase activity by up to 87%; the inhibition patterns suggest the presence of F0F1 and K+-ATPases but not Na+-, V- or Ca2+-ATPase as contaminants. (Na+, K+)-ATPase activity was synergistically modulated by K+ and NH4(+). At 10 mmol L(-1) K+, increasing NH4(+) concentrations stimulated maximum activity to V=185.9+/-7.4 U mg(-1). However, at saturating NH4(+) (50 mmol L(-1)), increasing K+ concentrations did not stimulate activity further. Our findings provide evidence that the C. ornatus gill (Na+, K+)-ATPase may be particularly well suited for extremely efficient active NH4(+) excretion. At elevated NH4(+) concentrations, the enzyme is fully active, regardless of hemolymph K+ concentration, and K+ cannot displace NH4(+) from its exclusive binding sites. Further, the binding of NH4(+) to its specific sites induces an increase in enzyme apparent affinity for K+, which may contribute to maintaining K+ transport, assuring that exposure to elevated ammonia concentrations does not lead to a decrease in intracellular potassium levels. This is the first report of modulation by ammonium ions of C. ornatus gill (Na+, K+)-ATPase, and should further our understanding of NH4(+) excretion in benthic crabs.     

K+-Phosphatase activity of gill (Na+, K+)-ATPase from the blue crab, Callinectes danae: low-salinity acclimation and expression of the alpha-subunit

The kinetic properties of a microsomal gill (Na(+), K(+)) ATPase from the blue crab, Callinectes danae, acclimated to 15 per thousand salinity for 10 days, were analyzed using the substrate p-nitrophenylphosphate. The (Na(+), K(+))-ATPase hydrolyzed the substrate obeying Michaelian kinetics at a rate of V=102.9+/-4.3 U.mg(-1) with K(0.5)=1.7+/-0.1 mmol.L(-1), while stimulation by magnesium (V=93.7+/-2.3 U.mg(-1); K(0.5)=1.40+/-0.03 mmol.L(-1)) and potassium ions (V=94.9+/-3.5 U.mg(-1); K(0.5)=2.9+/-0.1 mmol.L(-1)) was cooperative. K(+)-phosphatase activity was also stimulated by ammonium ions to a rate of V=106.2+/-2.2 U. mg(-1) with K(0.5)=9.8+/-0.2 mmol.L(-1), following cooperative kinetics (n(H)=2.9). However, K(+)-phosphatase activity was not stimulated further by K(+) plus NH(4) (+) ions. Sodium ions (K(I)=22.7+/-1.7 mmol.L(-1)), and orthovanadate (K(I)=28.1+/-1.4 nmol.L(-1)) completely inhibited PNPPase activity while ouabain inhibition reached almost 75% (K(I)=142.0+/-7.1 micromol.L(-1)). Western blotting analysis revealed increased expression of the (Na(+), K(+))-ATPase alpha-subunit in crabs acclimated to 15 per thousand salinity compared to those acclimated to 33 per thousand salinity. The increase in (Na(+), K(+))-ATPase activity in C. danae gill tissue in response to low-salinity acclimation apparently derives from the increased expression of the (Na(+), K( (+) ))-ATPase alpha-subunit; phosphate-hydrolyzing enzymes other than (Na(+), K(+))-ATPase are also expressed. These findings allow a better understanding of the kinetic behavior of the enzymes that underlie the osmoregulatory mechanisms of euryhaline crustaceans.     

Nitrophenylphosphate as a tool to characterize gill Na, K-ATPase activity in hyperregulating Crustacea

The kinetic properties of a gill Na(+), K(+)-ATPase from the freshwater shrimp Macrobrachium olfersii were studied using p-nitrophenylphosphate (PNPP) as a substrate. Sucrose gradient centrifugation of the microsomal fraction revealed a single protein fraction that hydrolyzed PNPP. The Na(+), K(+)-ATPase hydrolyzed PNPP (K(+)-phosphatase activity) obeying Michaelis-Menten kinetics with K(M)=1.72+/-0.06 mmol l(-1) and V(max)=259.1+/-11.6 U mg(-1). ATP was a competitive inhibitor of K(+)-phosphatase activity with a K(i)=50.1+/-2.5 micromol l(-1). A cooperative effect for the stimulation of the enzyme by potassium (K(0.5)=3.62+/-0.18 mmol l(-1); n(H)=1.5) and magnesium ions (K(0.5)=0.61+/-0.02 mmol l(-1), n(H)=1.3) was found. Sodium ions had no effect on K(+)-phosphatase activity up to 1.0 mmol l(-1), but above 80 mmol l(-1) inhibited the original activity by approximately 75%. In the range of 0-10 mmol l(-1), sodium ions did not affect stimulation of the K(+)-phosphatase activity by potassium ions. Ouabain (K(i)=762.4+/-26.7 micromol l(-1)) and orthovanadate (K(i)=0.25+/-0.01 micromol l(-1)) completely inhibited the K(+)-phosphatase activity, while thapsigargin, oligomycin, sodium azide and bafilomycin were without effect. These data demonstrate that the activity measured corresponds to that of the K(+)-phosphatase activity of the Na(+), K(+)-ATPase alone and suggest that the use of PNPP as a substrate to characterize K(+)-phosphatase activity may be a useful technique in comparative osmoregulatory studies of Na(+), K(+)-ATPase activities in crustacean gill tissues, and for consistent comparisons with well known mechanistic properties of the vertebrate enzyme.     

A kinetic study of the gill (Na+, K+)-ATPase, and its role in ammonia excretion in the intertidal hermit crab, Clibanarius vittatus

To better comprehend the role of gill ion regulatory mechanisms, the modulation by Na(+), K(+), NH(4)(+) and ATP of (Na(+), K(+))-ATPase activity was examined in a posterior gill microsomal fraction from the hermit crab, Clibanarius vittatus. Under saturating Mg(2+), Na(+) and K(+) concentrations, two well-defined ATP hydrolyzing sites were revealed. ATP was hydrolyzed at the high-affinity sites at a maximum rate of V=19.1+/-0.8 U mg(-1) and K(0.5)=63.8+/-2.9 nmol L(-1), obeying cooperative kinetics (n(H)=1.9); at the low-affinity sites, hydrolysis obeyed Michaelis-Menten kinetics with K(M)=44.1+/-2.6 mumol L(-1) and V=123.5+/-6.1 U mg(-1). Stimulation by Na(+) (V=149.0+/-7.4 U mg(-1); K(M)=7.4+/-0.4 mmol L(-1)), Mg(2+) (V=132.0+/-5.3 U mg(-1); K(0.5)=0.36+/-0.02 mmol L(-1)), NH(4)(+) (V=245.6+/-9.8 U mg(-1); K(M)=4.5+/-0.2 mmol L(-1)) and K(+) (V=140.0+/-4.9 U mg(-1); K(M)=1.5+/-0.1 mmol L(-1)) followed a single saturation curve and, except for Mg(2+), obeyed Michaelis-Menten kinetics. Under optimal ionic conditions, but in the absence of NH(4)(+), ouabain (K(I)=117.3+/-3.5 mumol L(-1)) and orthovanadate inhibited up to 67% of the ATPase activity. The inhibition studies performed suggest the presence of F(0)F(1), V- and P-ATPases, but not Na(+)-, K(+)- or Ca(2+)-ATPases as contaminants in the gill microsomal preparation. (Na(+), K(+))-ATPase activity was synergistically modulated by NH(4)(+) and K(+). At 20 mmol L(-1) K(+), a maximum rate of V=290.8+/-14.5 U mg(-1) was seen as NH(4)(+) concentration was increased up to 50 mmol L(-1). However, at fixed NH(4)(+) concentrations, no additional stimulation was found for increasing K(+) concentrations (V=135.2+/-4.1 U mg(-1) and V=236.6+/-9.5 U mg(-1) and for 10 and 30 mmol L(-1) NH(4)(+), respectively). This is the first report to detail ionic modulation of gill (Na(+), K(+))-ATPase in C. vittatus, revealing an asymmetrical, synergistic stimulation of the enzyme by K(+) and NH(4)(+), as yet undescribed for other (Na(+), K(+))-ATPases, and should provide a better understanding of NH(4)(+) excretion in pagurid crabs.     

Long-term exposure of the freshwater shrimp Macrobrachium olfersii to elevated salinity: effects on gill (Na+,K+)-ATPase alpha-subunit expression and K+-phosphatase activity

The kinetic properties of a microsomal gill (Na+,K+)-ATPase from the freshwater shrimp, Macrobrachium olfersii, acclimated to 21 per thousand salinity for 10 days were investigated using the substrate p-nitrophenylphosphate. The enzyme hydrolyzed this substrate obeying cooperative kinetics at a rate of 123.6+/-4.9 U mg-1 and K0.5=1.31+/-0.05 mmol L-1. Stimulation of K+-phosphatase activity by magnesium (Vmax=125.3+/-7.5 U mg-1; K0.5=2.09+/-0.06 mmol L-1), potassium (Vmax=134.2+/-6.7 U mg-1; K0.5=1.33+/-0.06 mmol L-1) and ammonium ions (Vmax=130.1+/-5.9 U mg-1; K0.5=11.4+/-0.5 mmol L-1) was also cooperative. While orthovanadate abolished p-nitrophenylphosphatase activity, ouabain inhibition reached 80% (KI=304.9+/-18.3 micromol L-1). The kinetic parameters estimated differ significantly from those for freshwater-acclimated shrimps, suggesting expression of different isoenzymes during salinity adaptation. Despite the approximately 2-fold reduction in K+-phosphatase specific activity, Western blotting analysis revealed similar alpha-subunit expression in gill tissue from shrimps acclimated to 21 per thousand salinity or fresh water, although expression of phosphate-hydrolyzing enzymes other than (Na+,K+)-ATPase was stimulated by high salinity acclimation.     

Mechanisms behind Pb-induced disruption of Na+ and Cl- balance in rainbow trout (Oncorhynchus mykiss)

The mechanism of Pb-induced disruption of Na(+) and Cl(-) balance was investigated in the freshwater rainbow trout (Oncorhynchus mykiss). Na(+) and Cl(-) influx rates were reduced immediately in the presence of 2.40 +/- 0.24 and 1.25 +/- 0.14 muM Pb, with a small increase in efflux rates occurring after 24-h exposure. Waterborne Pb caused a significant decrease in the maximal rate of Na(+) influx without a change in transporter affinity, suggesting a noncompetitive disruption of Na(+) uptake by Pb. Phenamil and bafilomycin markedly reduced Na(+) influx rate but did not affect Pb accumulation at the gill. Time-course analysis in rainbow trout exposed to 0, 0.48, 2.4, and 4.8 microM Pb revealed time- and concentration-dependent branchial Pb accumulation. Na(+)-K(+)-ATPase activity was significantly reduced, with 4.8 microM exposure resulting in immediate enzyme inhibition and 0.48 and 2.4 microM exposures inhibiting activity by 24 h. Reduced activity was weakly correlated with gill Pb accumulation after 3- and 8-h exposures; this relationship strengthened by 24 h. Reduced Na(+) uptake was correlated with gill Pb burden after exposures of 3, 8, and 24 h. Immediate inhibition of branchial carbonic anhydrase activity occurred after 3-h exposure to 0.82 +/- 0.05 or 4.30 +/- 0.05 microM Pb and continued for up to 24 h. We conclude that Pb-induced disruption of Na(+) and Cl(-) homeostasis is in part a result of rapid inhibition of carbonic anhydrase activity and of binding of Pb with Na(+)-K(+)-ATPase, causing noncompetitive inhibition of Na(+) and Cl(-) influx.     

Copper induced alterations of biochemical parameters in the gill and plasma of Oreochromis niloticus

The main objective of this study was to determine the effects of copper exposure on copper accumulated in branchial tissue, gill Na+/K+-ATPase activity and plasma Na+, Cl-, osmolality, protein, glucose and cortisol, in Oreochromis niloticus. Fish were experimentally exposed to 40 and 400 microg L(-1) of waterborne copper and sacrified after 0, 3, 7, 14 and 21 days. Copper accumulation and Na+/K+-ATPase activity were determined in branchial tissue, whereas osmolality, Na+, Cl-, protein, glucose and cortisol concentrations were measured in plasma samples. Gill copper accumulation increased linearly with exposure time and concentration, whereas gill Na+/K+-ATPase activity was maximally inhibited after 3 days of exposure and showed a significant negative correlation with copper tissue levels. Plasma Cl- values decreased with time of exposure but only at 400 microg L(-1) of copper. Plasma Na+, protein and osmolality decreased with exposure time at the highest copper concentration tested, whereas at 40 microg L(-1) of copper this effect was only observed after 21 days of exposure. Plasma glucose and cortisol levels increased in a dose and time dependent manner, while showing complex fluctuations during the intermediate exposure times. In conclusion, copper induces an early maximum inhibition of gill Na+/K+-ATPase activity in O. niloticus. The subsequent slow decrease in ion plasma levels was related to compensatory mechanisms involving a non-specific stress response that appeared overcome at long-term exposures.     

Wild Arctic char (Salvelinus alpinus) upregulate gill Na+,K+-ATPase during freshwater migration

The successful acclimation of eurhyhaline fishes from seawater to freshwater requires the gills to stop actively secreting ions and start actively absorbing ions. Gill Na(+),K(+)-ATPase is known to be an integral part of the active ion secretion model of marine fishes, but its importance in the active ion uptake model of freshwater fishes is less clear. This study, conducted in the high Arctic, examines gill Na(+),K(+)-ATPase regulation in wild anadromous arctic char returning to freshwater from the ocean. Gill Na(+),K(+)-ATPase activity, protein expression, and mRNA expression of Na(+),K(+)-ATPase isoforms alpha 1a and alpha 1b were monitored in arctic char at three points along their migration route to and from Somerset Island, Nunavut, Canada: out at sea (Whaler's Point), in seawater near the river mouth (Nat's Camp), and after entering the Union River. Arctic char collected from the Union River had more than twofold greater gill Na(+),K(+)-ATPase activity. This was associated with a significant increase (threefold) in Na(+),K(+)-ATPase isoform alpha 1a mRNA expression and a significant increase in plasma sodium and osmolality levels compared with seawater char. Compared with char sampled from Whaler's Point, Na(+),K(+)-ATPase isoform alpha 1b mRNA expression was decreased by approximately 50% in char sampled at Nat's Camp and the Union River. These results suggest that the upregulation of gill Na(+),K(+)-ATPase activity is involved in freshwater acclimation of arctic char and implicate a role for Na(+),K(+)-ATPase isoform alpha 1a in this process. In addition, we discuss evidence that arctic char go through a preparatory phase, or "reverse smoltification," before entering freshwater.     

Tissue ATPase activity and recovery in the freshwater crab Oziotelphusa senex senex exposed to a sublethal concentration of endosulfan for varying periods of time.

1. Na(+)-K+ and Mg(2+)-tissue ATPases of the freshwater crab Oziotelphusa senex senex showed increasing inhibition when exposed to a sublethal concentration (1.86 mg/l = 0.1 of LC50) of endosulfan for 1-30 days. 2. Na(+)-K(+)-ATPase activity in all tissues (thoracic nerve mass, gill, hepatopancreas and claw muscle) was higher than Mg(2+)-ATPase activity. 3. After 30 days exposure tissue Mg(2+)-ATPase was less affected than Na(+)-K(+)-ATPase. 4. Crabs exposed to endosulfan and then returned to uncontaminated water showed greater recovery of Mg(2+)-ATPase than Na(+)-K(+)-ATPase with 90-95% recovery after 1 day exposure and 60-65% recovery after 30 days exposure. 5. Changes in behaviour of the crabs were noted after 7 days exposure to endosulfan with progressive loss of coordination, decreased activity and increased exudation of mucus.