盐度、温度对卵形鲳鲹选育群体肝抗氧化酶活力的影响

通过盐度渐变和温度骤变的方法,分别研究了不同盐度(10、20、30、40)处理和不同温度(18.0℃、21.0℃、24.6℃、29.0℃、32.0℃)处理对卵形鲳鲹(Trachinotus ovatus)选育群体肝超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GPX)活力的影响.在实验结束时,盐度10组酶活力与对照组无显著差异(P＞0.05),盐度20组SOD活力极显著低于对照组(P＜0.01);盐度40组120 h时,SOD和GPX活力极显著低于对照组(P＜0.01),CAT活力与对照组差异不显著(P＞0.05).18.0℃和21.0℃ SOD活力在1、3、6、12、24 h这5个取样时间点均高于对照组,CAT活力在实验结束时(24 h)极显著高于对照组(P ＜0.01);29.0℃SOD和CAT活力在实验结束时(24 h)显著高于对照组(P＜0.05);32.0℃ SOD和CAT活力在5个取样时间点均显著低于对照组(P＜0.05).结果表明,适当的盐度或温度变化可以改变卵形鲳鲹肝抗氧化酶活力,达到机体耐受极限时酶活力下降.     

高胆固醇血症病人的红细胞膜ATP酶活性变化

研究表明高胆固醇血症病人的红细胞膜Na⁺-K⁺-ATP酶和Ca²⁺-Mg²⁺-ATP酶活性均降低,并且血浆总胆固醇和低密度脂蛋白-胆固醇浓度与这两种酶活性呈高度负相关,而血浆高密度脂蛋白-胆固醇浓度与Na⁺-K⁺-ATP酶活性呈正相关。这些变化的研究,对于进一步探讨动脉粥样硬化的发生机制及其防治可能具有重要意义。     

急性和慢性低氧胁迫对卵形鲳鲹幼鱼肝组织损伤和抗氧化的影响

自然海域和养殖水体环境频现低氧,本研究针对卵形鲳鲹(Trachinotus ovatus)不耐低氧的特性,将(31.59±3.01)g(n=30)的卵形鲳鲹在(23±0.7)℃下进行3、6、12和24 h的急性和14 d的慢性低氧[溶解氧为(1.55±0.20)mg/L]胁迫。运用光学和电子显微技术,比较急、慢性低氧胁迫对卵形鲳鲹肝组织显微和超微结构的影响。通过测定肝组织中的丙二醛(MDA)含量及过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和谷胱甘肽还原酶(GSH)的活性,分析低氧胁迫对卵形鲳鲹肝是否造成氧化损伤。急性低氧胁迫下,卵形鲳鲹肝组织间出现空泡、小叶结构破坏,细胞内线粒体数量减少,出现过氧化物酶体,肝细胞间血窦剧烈扩张。这些病理损伤随胁迫时间延长更趋严重,24 h时甚至出现局部肝细胞融合、坏死,慢性低氧胁迫14 d时,肝细胞局部坏死,细胞膜溶解,细胞核破裂分解,胞质内细胞器不明显,只可分辨粗面内质网,细胞内空泡体积大,细胞结构松散,血窦扩张。急性低氧胁迫下丙二醛(MDA)含量随时间先上升后下降,慢性低氧胁迫14 d时丙二醛(MDA)则显著增加。急性低氧胁迫下超氧化物歧化酶(SOD)和谷胱甘肽还原酶(GSH)上升后恢复,过氧化氢酶(CAT)则持续上调,慢性低氧胁迫下超氧化物歧化酶(SOD)和谷胱甘肽还原酶(GSH)显著上调,过氧化氢酶(CAT)活性下降。结果表明,卵形鲳鲹幼鱼肝组织在低氧胁迫下病理变化明显,氧化损伤严重,且慢性低氧胁迫比急性更甚。     

不同季节荷斯坦牛3个微卫星座位与GSH-Px、SOD和Na+/K+-ATP酶活性及日产奶量的关系

选择分别与谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)和Na⁺/K⁺-ATP酶基因紧密连锁的3个微卫星座位BMS2258、SOD1和BM723, 采用PCR及非变性聚丙烯酰胺凝胶电泳分析其在130头荷斯坦牛中的遗传变异, 计算了3个微卫星座位的多态信息含量、有效等位基因数和遗传杂合度, 并利用最小二乘法拟合线性模型初步探索了它们与荷斯坦牛夏、秋季GSH-Px、SOD、Na⁺/K⁺-ATP酶活性及日产奶量的关系。结果表明, 3个微卫星座位与其紧密连锁基因的酶活性及日产奶量均存在显著相关(P<0.05)。BMS2258座位182 bp/164 bp的GSH-Px活性和日产奶量对应的最小二乘均值较高; SOD1座位148 bp/146 bp对应的SOD活性的最小二乘均值较高, 148 bp/148 bp对应的日产奶量最小二乘均值较高; BMS2258座位161 bp/111 bp对应的Na⁺/K⁺-ATP酶活性和日产奶量的最小二乘均值较高, 它们是各自座位上的最有利基因型。     

环境因子对卵形鲳鲹幼鱼耗氧率和排氨率的影响

运用封闭流水式实验方法研究温度、盐度、pH和流速对卵形鲳鲹(Trachinotus ovatus)幼鱼耗氧率和排氨率的影响.实验结果表明,随着温度的升高,耗氧率和排氨率均是先增大后减小,当温度为27℃时,耗氧率和排氨率达最大值,温度对卵形鲳鲹幼鱼耗氧率和排氨率的影响显著(P＜0.0l);耗氧率和排氨率随着盐度的升高均出...     

急性盐度胁迫对军曹鱼稚鱼渗透压调节的影响

研究了环境盐度急性胁迫对军曹鱼(Rachycentron canadum)稚鱼鳃Na⁺-K⁺ATPase(NKA)活性及血清渗透压、Na⁺、K⁺和Cl^-离子调节的影响.结果表明:将稚鱼从盐度37中直接转移至盐度0、5、15、25、37（对照）和45的水体中,12 h后仅盐度0处理出现死亡(死亡率100％).各处理鳃NKA活性和血清渗透压在最初3 h内出现一定波动,随后变化平稳.试验结束时(12 h), NKA活性与盐度梯度呈“U”型分布,盐度5处理酶活性显著高于其它处理(P<0.05),盐度15处理活性最低,而各处理的血清渗透压大小(293～399 mOsmol·kg^-1)与盐度呈正相关;在3～12 h内稚鱼血清Na⁺和Cl^-浓度随盐度升高而升高,但增幅较小,血清K⁺浓度则与盐度呈负相关;12 h稚鱼的等渗点为328.2 mOsm·kg^-1,相当于盐度11.48,而Na⁺、K⁺和Cl^-等离子点分别为155.2、6.16和137.1 mmol·L^-1,分别相当于盐度10.68、20.44及8.41.军曹鱼在生理上具有广盐性鱼类的“低渗环境高NKA活性”特征,有较强及迅速的渗透压和离子调节与平衡能力.     

卵形鲳鲹幼鱼耗氧率和排氨率的初步研究

运用封闭流水式实验方法对不同体重和放养密度的卵形鲳鲹(Trachinoms ovatus)幼鱼耗氧率和排氨率进行了初步的研究。结果表明,卵形鲳鲹幼鱼的耗氧率随着鱼体重的增加而逐渐下降,排氨率总体上也呈降低的趋势,体重对卵形鲳鲹耗氧率、排氨率的影响均达到显著水平(P0.05);光照对卵形鲳鲹幼鱼的耗氧率和排氨率影响非常显著(P0.01),幼鱼在遮光条件下的耗氧率和排氨率分别比在自然光照条件下要低25.18%～40.76%和16.28%～40.28%;随着放养密度的增加,卵形鲳鲹幼鱼的耗氧率逐渐降低,排氨率随着密度的增加呈先上升后下降的趋势,放养密度对卵形鲳鲹耗氧率、排氨率的影响均达到显著水平(P0.05)。卵形鲳鲹的耗氧率和排氨率具有明显的昼夜变化,白天的耗氧率和排氨率均高于夜间值,耗氧率的低谷值为高峰值的69.68%,排氨率的低谷值为高峰值的30.91%。卵形鲳鲹幼鱼的窒息点溶解氧含量为(0.991±0.058)mg/L。     

大鼠前脂细胞质膜Na+/H+交换同时参与细胞的增殖和分化

以原代培养的大鼠前脂细胞为模型 ,以 2′ ,7′ bis ( 2 carboxyethyl) 5 ( 6 ) carboxyfluorescein (BCECF)作为检测胞内pH(pHi)的荧光探针 ,测定不同生长因子刺激下胞内pH的变化 ,证明大鼠肾周前脂细胞质膜存在Na⁺/H⁺交换活性 ,胎牛血清(FCS)能快速激活Na⁺/H⁺交换 ,导致pHi升高 (约 0 .2pH单位 ) ,并引起DNA合成 .Ethyl isopropyl amiloride (EIPA)抑制Na⁺/H⁺交换与DNA合成 .在无血清条件下 ,胰岛素不刺激DNA合成但引起细胞分化 ,表现为胞内脂滴积累和 3 磷酸 甘油脱氢酶(G₃ PDH酶 )活性增强 ,同时激活Na⁺/H⁺交换活性导致pHi升高 ;EIPA既抑制胰岛素对Na⁺/H⁺交换的激活 ,也抑制G₃ PDH酶活性增强 .结果证明 :Na⁺/H⁺交换的激活不仅与大鼠前脂细胞增殖相关 ,同时也是细胞分化的早期事件 .     

短期NaCl胁迫对不同小麦品种幼苗K+吸收和Na+、K+积累的影响

为研究盐胁迫下小麦幼苗生长及Na⁺、K⁺的吸收和积累规律,以中国春、洲元9369和长武134等3种耐盐性不同小麦品种为材料,采用非损伤微测技术检测盐胁迫2 d后的根系K⁺离子流变化,并对植株体内的Na⁺、K⁺含量进行测定。结果表明:短期(2d)盐胁迫对小麦生长有抑制作用,且对根系的抑制大于地上部,耐盐品种下降幅度小于盐敏感品种。盐胁迫下,小麦根际的 K⁺大量外流,盐敏感品种中国春K⁺流速显著高于耐盐品种长武134,最高可达15倍。小麦幼苗地上部分和根系均表现为Na⁺积累增加,K⁺积累减少,Na⁺/K⁺比随盐浓度增加而上升。中国春限Na⁺能力显著低于长武134,Na⁺/K⁺则显著高于长武134。综上所述,盐胁迫下造成小麦组织器官中Na⁺/K⁺比上升的主要原因是根系K⁺大量外流和Na⁺的过量积累,耐盐性不同的小麦品种间差异显著,并认为根系对K⁺的保有能力可能是作物耐盐性评价的一个重要指标。     

温度和盐度对褐牙鲆幼鱼渗透生理及抗氧化水平的影响

采用双因素析因实验设计方法,研究了温度（20℃、24℃、28℃）和盐度（10、30）急性应激对褐牙鲆（Paralichthys olivaceus）幼鱼渗透生理和抗氧化水平的影响。结果表明:盐度和温度变化对各实验组1d和6d时褐牙鲆幼鱼血浆皮质醇含量均无显著性差异。在高温低盐（28℃、10）环境中1d时渗透压显著高于其他各实验组,6d时无显著性差异。牙鲆幼鱼在28℃环境中1d时鳃Na+-K+-ATP酶活性显著高于20℃和24℃;6d时,温度和盐度对牙鲆幼鱼鳃Na+-K+-ATP酶活性有显著交互影响作用。1d时,随着温度的升高或盐度的降低牙鲆幼鱼肝脏超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性呈现上升趋势,并且高温低盐（28℃、盐度10）组褐牙鲆幼鱼肝脏丙二醛（MDA）含量显著高于其他各组;在3个实验温度下,10环境中牙鲆幼鱼肝脏脂质过氧化物（LPO）的含量高于30。在6d时,各实验组间肝脏SOD、CAT活性及MDA含量无显著性差异。因此,褐牙鲆能够耐受温度2028℃和低至盐度10的环境条件,应激早期温度和盐度的变化可引起褐牙鲆幼鱼渗透生理和抗氧化水平的变化,高温低盐对褐牙鲆幼鱼抗氧化水平的影响最大,至6d可基本恢复稳定。       

温度和盐度对吉富品系尼罗罗非鱼幼鱼鳃Na+-K+-ATPase活力的联合效应

试验采用中心组合设计(central composite face-centered design,CCF)和响应曲面法(response surface methodology,RSM)研究了温度(12—34℃)和盐度(0—26)两因素对体长为(4.36±0.105)cm,体重为(2.45±0.153)g的吉富品系尼罗罗非鱼(GIFT Nile tilapia,Oreochromis niloticus;简称吉富罗非鱼)幼鱼鳃Na+-K+-ATPase活力的联合效应。结果表明:(1)温度和盐度的一次效应和二次效应对Na+-K+-ATPase活力影响极显著(P<0.01),温度和盐度的互作效应不显著(P>0.05);(2)经响应曲面法分析,随着温度和盐度的增大,Na+-K+-ATPase活力呈先减小后增大的趋势;(3)建立了Na+-K+-ATPase活力与温度、盐度间关系的模型方程(R2=0.9829,Pred.R2=0.8550,P<0.01),并可用于预测吉富罗非鱼幼鱼鳃Na+-K+-ATPase的活力;(4)优化结果显示,温度为24.15℃,盐度为11.75时,Na+-K+-ATPase活力最小为0.62μmol无机磷.mg-1蛋白.h-1,满意度函数值高达0.961。Na+-K+-ATPase活力可以作为检测罗非鱼生长性能的指标,其活力较低时,一般反映了鱼体生存环境适宜,生长代谢旺盛,消耗于渗透调节的能量较少。     

Response to environmental salinity of Na-KATPase activity in individual gills of the euryhaline crab Cyrtograpsus angulatus

The occurrence, localization and response to environmental salinity changes of Na⁺-K⁺ATPase activity were studied in each of the individual gills 4-8 of the euryhaline crab Cyrtograpsus angulatus from Mar Chiquita coastal lagoon (Buenos Aires Province, Argentina). Na⁺-K⁺ATPase activity appeared to be differentially sensitive to environmental salinity among gills. Upon an abrupt change to low salinity, a differential response of Na⁺-K⁺ATPase activity occurred in each individual gill which could suggest a differential role of this enzyme in ion transport process in the different gills of C. angulatus. With the exception of gill 8, a short-term increase of Na⁺-K⁺ATPase specific activity was observed in posterior gills, which is similar to adaptative variations of this activity described in other euryhaline crabs. However, and conversely to that described in other hyperregulating crabs, the highest increase of activity occurred in anterior gills 4 by 1 day after the change to dilute media which could suggest also a role for these gills in ion transport processes in C. angulatus. The fact that variations of Na⁺-K⁺ATPase activity in anterior and posterior gills were concomitant with the transition to hyperregulation indicate that this enzyme could be a component of the branchial ionoregulatory mechanisms at the biochemical level in this crab. The results suggest a differential participation of branchial Na⁺-K⁺ATPase activity in ionoregulatory mechanisms of C. angulatus. The possible existence of functional differences as well as distinct regulation mechanisms operating in individual gills is discussed.     

Survival,osmoregulation and ammonia-N excretion of blue swimmer crab, Portunus pelagicus, juveniles exposed to different ammonia-N and salinity combinations

Ammonia-N toxicity to early Portunus pelagicus juveniles at different salinities was investigated along with changes to haemolymph osmolality, Na⁺, K⁺, Ca²⁺ and ammonia-N levels, ammonia-N excretion and gill Na⁺/K⁺-ATPase activity. Experimental crabs were acclimated to salinities 15, 30 and 45‰ for one week and 25 replicate crabs were subsequently exposed to 0, 20, 40, 60, 80, 100 and 120 mg L^− 1 ammonia-N for 96-h, respectively. High ammonia-N concentrations were used to determine LC₅₀ values while physiological measurements were conducted at lower concentrations. When crabs were exposed to ammonia-N, anterior gill Na⁺/K⁺-ATPase activity significantly increased (p < 0.05) at all salinities, while this only occurred on the posterior gills at 30‰. For crabs exposed to 20 and 40 mg L^− 1 ammonia-N, both posterior gill Na⁺/K⁺-ATPase activity and ammonia-N excretion were significantly higher at 15‰ than those at 45‰. Despite this trend, the 96-h LC₅₀ value at 15‰ (43.4 mg L^− 1) was significantly lower (p < 0.05) than at both 30‰ and 45‰ (65.8 and 75.2 mg L^− 1, respectively). This may be due to significantly higher (p < 0.05) haemolymph ammonia-N levels of crabs at low salinities and may similarly explain the general ammonia-N toxicity pattern to other crustacean species.     

Effect of water temperature,salinity, and their interaction on growth,plasma osmolality,and gill Na,K-ATPase activity in juvenile GIFT tilapia Oreochromis niloticus (L.)

We used a central composite rotatable experimental design and response surface methodology to evaluate the effects of temperature (18–37 °C), salinity (0–20‰), and their interaction on specific growth rate (SGR), feed efficiency (FE), plasma osmolality, and gill Na⁺, K⁺-ATPase activity in GIFT tilapia juveniles. The linear and quadratic effects of temperature and salinity on SGR, plasma osmolality, and gill Na⁺, K⁺-ATPase activity were statistically significant (P<0.05). The interactive effects of temperature and salinity on plasma osmolality were significant (P<0.05). In contrast, the interaction term was not significant for SGR, FE, and gill Na⁺, K⁺-ATPase activity ⁽P>0.05). The regression equations for SGR, FE, plasma osmolality, and gill Na⁺, K⁺-ATPase activity against the two factors of interest had coefficients of determination of 0.944, 0.984, 0.966, and 0.960, respectively (P<0.01). The optimal temperature/salinity combination was 28.9 °C/7.8‰ at which SGR (2.26% d¹) and FE (0.82) were highest. These values correspond to the optimal temperature/salinity combination (29.1 °C/7.5‰) and the lowest plasma osmolality (348.38 mOsmol kg⁻¹) and gill Na⁺, K⁺-ATPase activity (1.31 µmol Pi. h⁻¹ g⁻¹ protein), and resulted in an energy-saving effect on osmoregulation, which promoted growth.     

Changes in the selected hematological parameters and gill Na/K ATPase activity of juvenile crucian carp Carassius auratus during elevated ammonia exposure and the post-exposure recovery

The increase in concentration of ammonia in lake water during the degradation of algal blooms may last for several weeks and thus cause chronic toxicity to aquatic organisms. The purpose of this study was to assess the chronic toxicity of ammonia on the selected hematological parameters and gill Na⁺/K⁺ ATPase activity of juvenile crucian carp Carassius auratus during elevated ammonia exposure and the post-exposure recovery. Juvenile crucian carp were exposed in different ammonia solutions for 45 days and then immediately transferred to pristine freshwater to initiate a 15-day recovery period. Results showed sub-lethal ammonia significantly deters growth and a 15-day recovery period was not sufficient for the fish to compensate for the loss of growth. The fish exhibited a continuous decrease in red blood cell (RBC), the total hemoglobin (Hb), and gill Na⁺/K⁺ ATPase activity as the concentration of NH₃-N increased. After the 15-day recovery period, RBC, Hb, and gill Na⁺/K⁺ ATPase activity had recovered to similar levels as the controls.     

A nhaD Na/Hantiporter and a pcd homologues are among the Rhodothermus marinus complex I genes

The NADH:menaquinone oxidoreductase (Nqo) is one of the enzymes present in the respiratory chain of the thermohalophilic bacterium Rhodothermus marinus. The genes coding for the R. marinus Nqo subunits were isolated and sequenced, clustering in two operons [nqo₁ to nqo₇ (nqo_A) and nqo₁₀ to nqo₁₄ (nqo_B)] and two independent genes (nqo₈ and nqo₉). Unexpectedly, two genes encoding homologues of a NhaD Na⁺/H⁺ antiporter (NhaD) and of a pterin-4α-carbinolamine dehydratase (PCD) were identified within nqo_B, flanked by nqo₁₃ and nqo₁₄. Eight conserved motives to harbour iron-sulphur centres are identified in the deduced primary structures, as well as two consensus sequences to bind nucleotides, in this case NADH and FMN. Moreover, the open-reading-frames of the putative NhaD and PCD were shown to be co-transcribed with the other complex I genes encoded by nqo_B. The possible role of these two genes in R. marinus complex I is discussed.     

不同饵料和饥饿对魁蚶幼虫生长和存活的影响

在水温23.2-24.0℃,盐度29.5-30.0条件下,研究了5种单胞藻饵料和饥饿对魁蚶浮游幼虫生长与存活的影响.投喂不同饵料试验中,球等鞭金藻与4种单胞藻混合投喂试验组幼虫的特定生长率、眼点幼虫比例和壳长均显著高于单一投喂试验组,其中球等鞭金藻与小球藻混合投喂的效果最好;投喂单一饵料试验中,球等鞭金藻组幼虫的生长、眼点幼虫比例与存活率显著高于其它试验组.在饥饿试验中,不同饥饿天数条件下幼虫的存活率差异不显著;生长方面,饥饿1-2d较短时间与一直投喂对照组的幼虫相比较,特定生长率、眼点幼虫比例、壳长的差异不显著,但随着饥饿时间延长,幼虫的生长变慢且眼点幼虫比例下降,一直饥饿情况下幼虫的生长基本停滞.研究结果可以为魁蚶人工苗种繁育技术的改进提供参考依据.     

Asp and Thr are critical for cation exchange mediated by NhaD, Na/H antiporter of Vibrio cholerae

The Vc-NhaD is an Na⁺/H⁺ antiporter from Vibrio cholerae belonging to a new family of bacterial Na⁺/H⁺ antiporters, the NhaD family. In the present work we mutagenized five conserved Asp and Glu residues and one conserved Thr residue to Ala in order to identify amino acids that are critical for the antiport activity. All mutations fall into two distinct groups: (i) four variants, Glu¹⁰⁰Ala, Glu²⁵¹Ala, Glu³⁴²Ala, and Asp³⁹³Ala, did not abolish antiport activity but shifted the pH optimum to more alkaline pH, and (ii) variants Asp³⁴⁴Ala, Asp³⁴⁴Asn, and Thr³⁴⁵Ala caused a complete loss of both Na⁺/H⁺ and Li⁺/H⁺ antiport activity whereas the Asp³⁴⁴Glu variant exhibited reduced Na⁺/H⁺ and Li⁺/H⁺ antiport activity. This is the first mutational analysis of the antiporter of NhaD type and the first demonstration of Thr residue being indispensable for Na⁺/H⁺ antiport. We discuss the possible role of Asp³⁴⁴ and Thr³⁴⁵ in the functioning of Vc-NhaD.