盐度对银鲳(Pampus argenteus)Na /K -ATP酶活力及血清渗透压调节激素浓度的影响

采用检测银鲳(Pampus argenteus)幼鱼鳃Na+/K+-ATP(NKA)酶活力、血清生长激素(GH)、胰岛素样生长因子Ⅰ(IGF-Ⅰ)和催乳素(PRL)浓度的方法, 研究了不同盐度下银鲳渗透压调节机制。结果表明, 鳃NKA酶活力在三种盐度下总体呈先上升后恢复的变化, 其最高点为14盐度48h时, 且显著高于其它值(P<0.05)。36盐度组的血清GH浓度显著高于其它组(P<0.05)。盐度变化均使血清IGF-Ⅰ浓度呈现先上升后恢复的变化。14盐度组血清催乳素浓度始终高于其它各盐度组。在低盐度适应(14和25)时, 血清GH浓度与NKA酶活力呈极显著正相关(P<0.01); 高盐度适应(36和25)时, 血清催乳素浓度与NKA酶活力呈显著负相关(P<0.01)。另外, 血清催乳素与IGF-Ⅰ浓度间有极显著正相关(P<0.01)。研究证明银鲳幼鱼在适应不同盐度时, 鳃NKA酶、生长激素、IGF-Ⅰ和催乳素在渗透压调节中起着重要作用, 并且相互之间存在一定关联。     

Effect of various Na/K ratios in low-salinity well water on growth performance and physiological response of Pacific white shrimp Litopenaeus vannamei

To investigate the influence of sodium to potassium(Na/K) ratios on the growth performance and physiological response of the Pacific white shrimp( Litopenaeus vananmei), various concentrations of KCl were added to low-salinity well water(salinity 4) in an 8-week culture trial. Six treatments with Na/K ratios of 60:1, 42:1, 33:1, 23:1, 17:1, and 14:1 were replicated in triplicate. The highest weight-gain rate(3 506±48)% and survival rate(89.38±0.88)% was observed in well water with Na/K ratios of 23:1 and 42:1, respectively, while the feed conversion ratio(1.02±0.01), oxygen consumption, and ammonia-N excretion rate was the lowest in the medium with a Na/K ratio of 23:1. Gill Na +-K +-ATPase activity, as an indicator of osmoregulation, peaked in the treatment where the Na/K ratio was 17:1. The total hemocyte count, respiratory burst, and immune-related enzyme activities(ALP, LSZ, PO, and SOD) of L. vananmei were affected significantly by Na/K ratios( P 0.05). After challenged with Vibrio harveyi, the cumulative mortality of shrimp reared in a Na/K ratio of 23:1(30±14.14)% was significantly lower than the control(75±7.07)%. In conclusion, the addition of K + to low-salinity well water in L. vannamei cultures is feasible. Na/K ratios ranging from 23:1 to 33:1 might improve survival and growth. Immunity and disease resistance are also closely related to the Na/K ratio of the low-salinity well water. The findings may contribute to the development of more efficient K + remediation strategies for L. vananmei culture in low-salinity well water.     

凡纳滨对虾水孔蛋白-4的cDNA克隆及盐度胁迫对其肝胰腺mRNA表达水平的影响

为了探索水孔蛋白(Aquaporin,AQP)在凡纳滨对虾(Litopenaeus vannamei)渗透压调节过程中作用,本研究通过RACE方法获得克隆的凡纳滨对虾AQP(命名为LvAQP4)的c DNA全长序列,并分析了盐度胁迫对其肝胰腺m RNA表达的影响。结果发现:LvAQP4 c DNA序列全长为1 048 bp,其中包括75 bp的5￠UTR,187 bp的3￠UTR和786 bp的ORF。根据ORF序列推导出LvAQP4编码261个氨基酸,预测其分子质量为27.85 k Da,等电点为8.11。推导的氨基酸序列与其他甲壳物种的AQP相似度为48.8%到97.3%。进化分析显示LvAQP4属于AQP1-like亚族、AQP4类。定量PCR(q PCR)检测到LvAQP4在不同组织中均有表达,其中鳃的表达量最高,肝胰腺、肌肉、脑和眼柄中也较高,而血淋巴、肠、胃和胸神经节中表达量则较低。在高盐(盐度40)刺激下,凡纳滨对虾的肝胰腺LvAQP4 m RNA表达量会随着时间推移而上升,到6 h达到最高,而后逐步下降。而在低盐(盐度4)刺激下,凡纳滨对虾的肝胰腺LvAQP4 m RNA表达量并无明显变化。在原代分离和培养的肝胰腺细胞中,培养液中加入额外的Na Cl会剂量依赖地提高LvAQP4 m RNA表达水平。同样,通过在培养液中额外加入蔗糖提高培养液渗透压,也会剂量依赖地提高LvAQP4 m RNA表达水平,但作用不如Na Cl明显。这些结果表明盐度与肝胰腺LvAQP4的表达量有关,LvAQP4对凡纳滨对虾的渗透压调节具有非常重要的作用。     

温度和手术对扁额原细首纽虫渗透压调节的影响

研究3个温度下(5℃、15℃、25℃),扁额原细首纽虫在外界渗透压变化时体内渗透压的的变化以及手术(切头和中切)对其渗透压调节能力的影响。结果显示,温度对扁额原细首纽虫的渗透压调节具有较大影响。在25℃、96 h内,当纽虫处于低渗环境(盐度10和20)时,其体内渗透压先降后升,当其处于高渗环境(盐度40)时,其体内渗透压先升后降,表现为1个渗透压调节者。在15℃和5℃,纽虫的体内渗透压基本上是随着环境的渗透压变化而变化,表现为1个渗透压随变者。手术对扁额原细首纽虫的渗透压调节能力具有明显的影响。去头纽虫丧失了调节渗透压的能力,其体内渗透压随着环境渗透压变化而变化。当纽虫被中切为两段时,具有头部的身体前部能像完整纽虫一样调节其体内渗透压,而身体后段的渗透压随环境的渗透压变化而变化,没有明显的调节能力。结果显示头部器官(脑神经节)对这种纽虫维持渗透压调节能力是必要的,该纽虫可能通过神经内分泌来调控其渗透压调节。     

金钱鱼肾细胞系的建立及生长特性研究

金钱鱼(Scatophagus argus)是一种重要的广盐性海水养殖鱼类,可直接在海水、淡水、咸淡水等不同环境中正常生长。为研究金钱鱼独特的渗透调节机制,本文研究了金钱鱼肾细胞的原代和传代条件及其生长特性,结果表明:原代肾细胞在含有20%胎牛血清(FBS)的L-15培养基里贴壁和生长较好,添加10ng/mL的碱性成纤维细胞因子(basic fibroblast growth factor)bFGF能明显促进细胞增殖,传代细胞采用含有10%胎牛血清的L-15培养基,细胞生长迅速,3—4d即可传代。金钱鱼肾细胞为成纤维样细胞,命名为SK细胞,目前已传至22代。扩增第11代SK细胞的细胞色素氧化酶I(COI)基因,比对结果证明此细胞系来源于金钱鱼。采用CCK-8法检测第12代细胞在低渗(95,137,200mmol/kg),等渗(330mmol/kg)和高渗(430,550mmol/kg)中的增殖情况,结果发现肾细胞在137—430mm/kg的渗透压范围内均可增殖,说明金钱鱼肾细胞对渗透压的耐受性较强。本试验首次建立了金钱鱼肾细胞系,并初步证明了肾细胞对盐度有较强的耐受性,为今后渗透压调节的深入研究奠定了基础。     

Impacts of Habitat Complexity on Physiology: Purple Shore Crabs Tolerate Osmotic Stress for Shelter

Purple shore crabs, Hemigrapsus nudus (Crustacea: Decapoda), can survive indefinitely in salinities of 8 (25% SW), but also tolerate short-term exposure to salinities as low as 2. In the laboratory their salinity preference range, determined from choice chamber experiments, is 22–32 and they can discriminate between pairs of salinities separated by a difference of 2. These crabs show a strong positive thigmotaxis and a weak negative phototaxis and tend to choose environments with available shelter. The presence of shelter significantly alters the behaviour of this species. When shelter is available the salinity preference range is 10–32. Even in salinities below this preference range, the presence of shelter prolongs the time spent in the lower test salinities. This change in behaviour has implications on the crab's physiology: the haemolymph osmolality falls to lower levels when crabs remain in low salinity under shelters. In the field, H. nudus is found in creeks with salinities close to freshwater and they may remain in this salinity for up to 11 h, if there are rocks under which to shelter. An increase in habitat complexity increases the number of crabs that are found within the creek. These crabs in the low salinity environment have a lower haemolymph osmolality than crabs on the nearby open shore. In H. nudus the behavioural selection of a shelter appears to outweigh the physiological costs associated with osmotic regulation of the body fluids. Therefore, the distribution of H. nudus in estuaries may depend more on the availability of suitable habitats rather than the salinity tolerance of this species.     

沙漠植物抗氧化酶活性及渗透调节物质含量与光合作用的关系

以科尔沁沙地生长的沙漠植物差不嘎蒿(Artimisia halodendron)、小叶锦鸡儿(Caragana microphylla)、扁蓄豆(Ruthenian medic)、胡枝子(Lespedeza bicolor)、黄柳(Salix gordejevii)和猪毛菜(Salsola collina)为材料,研究了自然状况下光合作用、保护酶活性及渗透调节物质含量的日变化特征及相关关系。结果表明:(1)5个C₃植物光合作用日变化为中午降低类型,净光合速率(P_n)在10:00到达第1个高峰,在14:00出现第2个高峰,其中4个植物第2高峰平均P_n仅为第1高峰的37%。C₄植物猪毛菜光合作用为单峰型。(2)在第1个P_n高峰期间,伴随着光合有效辐射(PAR)增高6.5倍和气温(T_a)增高,叶片水分亏缺度(VPD)增加3~6倍、气孔导度(G_s)下降60%;叶片丙二醛(MDA)含量增加18%,P_n增加3~13倍;过氧化物酶(POD)活性(除了扁蓄豆)平均增高29%、过氧化氢酶(CAT)活性(除胡枝子)平均增加82%;叶片可溶性糖含量增加48%,脯氨酸含量(除差不嘎蒿)增加74%。(3)午间和午后在大气PAR和T_a较高、叶片VPD较大、MDA含量和P_n下降时,渗调节物含量和抗氧化酶活性仍维持较高水平。日间叶片光合特性与保护酶活性和渗透调节物质含量的日变化间具有一定的相关性。抗氧化酶(尤其是POD和CAT)和渗透调节物(脯氨酸和可溶性糖)不仅在P_n增加过程中维护细胞氧自由基代谢和水分代谢平衡,而且在P_n较低时抑制膜脂过氧化和维持水分平衡,因此它们在日间提高P_n和恢复叶片光合作用上起重要的生理保护作用。6种植物参与抗逆生理调节的抗氧化酶和渗透调节物种类上差异显著。