盐碱胁迫对尼罗罗非鱼鳃Na+/HCO3-共转运子、碳酸酐酶基因表达的影响

为了探讨盐碱胁迫条件下鱼类渗透生理调节机制,以尼罗罗非鱼(Oreochromis niloticus)为实验材料, PCR扩增得到了Na⁺/HCO₃^-共转运子(NBCe1)基因cDNA部分序列,比较了单盐(盐度10、盐度15)、单碱(1.5 g/L、3 g/L NaHCO₃)、盐碱混合(盐度10,碱度1.5 g/L;盐度15,碱度3 g/L)胁迫后不同时间(0 h、6 h、12 h、24 h、48 h、72 h、96 h)血清渗透压、离子浓度(Na⁺、K⁺、Cl^-、Ca²⁺)以及鳃碳酸酐酶(CA)活性、CA与NBCe1基因mRNA表达变化。结果显示,不同胁迫条件下,血清渗透压、离子浓度、鳃组织CA酶活、CA与NBCe1基因mRNA表达变化均与胁迫强度呈正相关。随时间推移,血清渗透压、离子浓度呈现先上升后下降的变化趋势,单盐、盐碱混合组血清渗透压值较单碱组高。单盐、单碱、盐碱混合组中, NBCe1基因mRNA在鳃中均呈略微上调,但不显著(P>0.05)。单碱组和盐碱混合组鳃CA活性较单盐组高,低盐碱胁迫(盐度10,碱度1.5 g/L)下CA活性较晚达最高值;不同胁迫条件下, CA基因mRNA表达均表现上调,单碱、盐碱混合组更为显著(P<0.05),推测CA较NBCe1对体内HCO₃^-转运作用更为显著。研究结果为尼罗罗非鱼盐碱适应生理调节提供了基础资料。     

盐碱胁迫对尼罗罗非鱼鳃Na+/HCO3-共转运子、碳酸酐酶基因表达的影响

为了探讨盐碱胁迫条件下鱼类渗透生理调节机制,以尼罗罗非鱼(Oreochromis niloticus)为实验材料, PCR扩增得到了Na+/3HCO-共转运子(NBCe1)基因cDNA部分序列,比较了单盐(盐度10、盐度15)、单碱(1.5 g/L、3 g/L NaHCO3)、盐碱混合(盐度10,碱度1.5 g/L;盐度15,碱度3 g/L)胁迫后不同时间(0 h、6 h、12 h、24 h、48 h、72 h、96 h)血清渗透压、离子浓度(Na+、K+、Cl–、Ca2+)以及鳃碳酸酐酶(CA)活性、CA与NBCe1基因mRNA表达变化。结果显示,不同胁迫条件下,血清渗透压、离子浓度、鳃组织 CA 酶活、CA 与 NBCe1基因 mRNA 表达变化均与胁迫强度呈正相关。随时间推移,血清渗透压、离子浓度呈现先上升后下降的变化趋势,单盐、盐碱混合组血清渗透压值较单碱组高。单盐、单碱、盐碱混合组中, NBCe1基因mRNA在鳃中均呈略微上调,但不显著(P>0.05)。单碱组和盐碱混合组鳃CA活性较单盐组高,低盐碱胁迫(盐度10,碱度1.5 g/L)下CA活性较晚达最高值;不同胁迫条件下, CA基因mRNA表达均表现上调,单碱、盐碱混合组更为显著(P<0.05),推测CA较NBCe1对体内3HCO-转运作用更为显著。研究结果为尼罗罗非鱼盐碱适应生理调节提供了基础资料。     

3种品系尼罗罗非鱼(Oreochromis niloticus)盐碱耐受性和生长比较

为评估尼罗罗非鱼的耐盐碱性能,分别测定了上海、山东、河北3种品系尼罗罗非鱼鱼种96 h的半致死盐、碱度,并在不同盐碱混合浓度(S0A0、S10A0、S10A2、S10A4、S10A6)中进行为期60 d的养殖生长比较。单盐、单碱耐性研究表明,上海、山东、河北3种品系尼罗罗非鱼鱼种的96 h半致死盐度分别为18.528 g/L、20.347 g/L、19.342 g/L,96 h半致死Na HCO3碱度为8.827g/L、8.540 g/L、8.542 g/L。盐碱混合条件下,盐度为10时,96 h半致死碱度分别为河北品系(4.377g/L)上海品系(3.561 g/L)山东品系(3.108 g/L),品系之间差异显著(P0.05);盐度为15时,96 h半致死碱度分别为河北品系2.144 g/L,上海品系2.183 g/L,山东品系2.183 g/L,品系之间无显著差异(P0.05),高盐度下尼罗罗非鱼鱼种的碱度耐受性明显低于低盐度下的碱度耐受性。结果表明,尼罗罗非鱼日均增重率在S0A0、S10A0组间无显著差异(P0.05),随着盐碱浓度增加,盐碱S10A4和S10A6组中日均增重率呈下降趋势,河北品系表现出相对生长优势(P0.05)。研究结果为尼罗罗非鱼适宜养殖的盐碱范围的确定、耐盐碱品系的筛选提供了基础资料。     

3种品系尼罗罗非鱼(Oreochromis niloticus)盐碱耐受性和生长比较

为评估尼罗罗非鱼的耐盐碱性能,分别测定了上海、山东、河北3种品系尼罗罗非鱼鱼种96 h的半致死盐、碱度,并在不同盐碱混合浓度(S0A0、S10A0、S10A2、S10A4、S10A6)中进行为期60 d的养殖生长比较。单盐、单碱耐性研究表明,上海、山东、河北3种品系尼罗罗非鱼鱼种的96 h半致死盐度分别为18.528 g/L、20.347 g/L、19.342 g/L,96 h半致死Na HCO3碱度为8.827g/L、8.540 g/L、8.542 g/L。盐碱混合条件下,盐度为10时,96 h半致死碱度分别为河北品系(4.377g/L)上海品系(3.561 g/L)山东品系(3.108 g/L),品系之间差异显著(P0.05);盐度为15时,96 h半致死碱度分别为河北品系2.144 g/L,上海品系2.183 g/L,山东品系2.183 g/L,品系之间无显著差异(P0.05),高盐度下尼罗罗非鱼鱼种的碱度耐受性明显低于低盐度下的碱度耐受性。结果表明,尼罗罗非鱼日均增重率在S0A0、S10A0组间无显著差异(P0.05),随着盐碱浓度增加,盐碱S10A4和S10A6组中日均增重率呈下降趋势,河北品系表现出相对生长优势(P0.05)。研究结果为尼罗罗非鱼适宜养殖的盐碱范围的确定、耐盐碱品系的筛选提供了基础资料。     

黄斑蓝子鱼LC-PUFA 合成代谢与渗透压调节的关系研究

为探讨黄斑蓝子鱼(Siganus canaliculatus)长链多不饱和脂肪酸(long-chain polyunsaturated fatty acids,LC-PUFA)合成代谢与渗透压调节的关系,本研究以鱼油(FO)和混合植物油(苏子油与双低菜籽油,VO)为脂肪源配制两种等氮等脂饲料,投喂饲养在3种盐度(10、20和32)下的黄斑蓝子鱼幼鱼8周后,分析了各处理组幼鱼的生长性能和鳃的磷脂脂肪酸组成、Na+/K+-ATPase(NKA)活力及其基因表达。结果显示,相同盐度下,VO组和FO组鱼的生长性能差异不显著(P0.05);FO组鱼鳃磷脂中的n-3 LC-PUFA含量显著高于VO组(P0.05),但VO组鱼的n-6 LC-PUFA水平显著高于FO组(P0.05);VO组鱼鳃的NKA酶活力及其m RNA表达量都显著高于FO组(P0.05)。不同盐度下,无论VO组还是FO组的鱼,盐度10组鱼的生长性能显著低于盐度20和32组(P0.05),而其鳃的LC-PUFA含量、NKA酶活力及其m RNA表达量都显著高于盐度20和32组(P0.05),各指标在后两个盐度组之间差异不显著(P0.05)。由此可见,盐度10对黄斑蓝子鱼具有一定的胁迫性,导致其生长性能较差。摄食鱼油脂肪源饲料,可以提高鱼鳃磷脂的n-3 LC-PUFA水平;而摄食植物脂肪源饲料时,鱼体可能通过自身合成的n-6LC-PUFA调控鳃的NKA基因表达及其酶活力以调节渗透压。     

急性盐度胁迫对摄食不同镁水平饲料鲈血清渗透压和离子水平以及鳃丝ATP酶活力的影响

为研究饲喂不同镁水平饲料后,鲈血清渗透压、离子水平和鳃丝ATP酶活力对急性盐度胁迫的反应,实验用镁水平为0.413 g/kg(D1,对照组)、1.042 g/kg(D2)、1.577 g/kg(D3)、1.991 g/kg(D4)的4种实验饲料,分别投喂初始平均体质量为(30±0.5)g的鲈。实验鱼在淡水循环养殖系统中饲养50 d后立即转入海水循环养殖系统,并在转入前(0 h)和转入后1、3、6、12、24 h分别采集鲈鳃丝和血液样本。结果显示:盐度胁迫前饲料镁水平对鲈血清渗透压、Na+、K+、Cl-、Mg2+含量和鳃丝中Na+/K+-ATP(NKA)与Ca2+/Mg2+-ATP(CMA)酶的活力有显著影响(P<0.05),其中血清渗透压、Mg2+含量和NKA酶活力随饲料镁水平的增加而升高。D4组的血清渗透压、Mg2+含量、NKA和CMA酶活力显著高于其他组(P<0.05),表明该组鲈处于高强度的渗透压调节的生理状态。急性盐度胁迫中,血清Na+和Cl-含量随胁迫时间的延长呈逐渐上升趋势,血清渗透压、K+、Ca2+、Mg2+和鳃丝ATP酶活力在0～24 h内呈波动性变化。D1组的NKA和CMA酶活力在胁迫1 h时显著低于其他组(P<0.05),表明长期摄食低镁水平的饲料会降低鲈鳃丝NKA和CMA酶对环境盐度刺激的敏感度。研究表明,淡水养殖鲈的饲料中镁水平应低于1.991 g/kg,以减少维持鳃丝NKA与CMA酶高活力而导致的能量损失,而在环境由低盐向高盐的快速转变过程中,较高的饲料镁水平可通过快速提高的鳃丝NKA与CMA酶活力而有利于鲈快速适应高盐环境。     

中华鲟幼鱼鳃丝Na+,K+-ATPase α 亚基渗透调节的分子机制初步研究

为了研究盐度10条件下中华鲟幼鱼鳃丝Na+,K+-ATPase(NKA)α亚基的分子调节机制,采用克隆方法得到中华鲟幼鱼鳃丝NKAα亚基部分基因序列,在鲟鱼转移后3,6,12,24,48,72和96 h检测实验组(淡水-盐水)和对照组(淡水-淡水)中华鲟幼鱼鳃丝NKAα亚基的mRNA表达水平、鳃丝NKA活性、血清Na+、Cl-的浓度和渗透压。结果显示,在转移后0～12 h,实验组中的mRNA表达量与酶活性为适应盐度变化而显著增加(P<0.05),离子浓度和渗透压均显著上升(P<0.05);在转移后12～24 h,mRNA表达和酶活性开始降低到一个较低值,但仍然显著高于对照组的值(P<0.05),离子﹑渗透压的变化与酶活性及mRNA变化趋势一致,这表明中华鲟鳃丝NKA在应对盐度变化时通过改变mRNA表达量来增加酶活性,高活性NKA调节血清离子和渗透压平衡。     

碱胁迫下罗非鱼鳃氨转运蛋白Rh基因的表达

为研究碱环境下广盐性鱼类氨转运途径,将尼罗罗非鱼同时进行急性碱度胁迫(2、4、6 g/L)和慢性碱度胁迫,检测胁迫后120 h内的血氨浓度与鳃组织中Rhag、Rhbg、Rhcg1和Rhcg2基因mRNA表达变化,并采用免疫组化技术观察鳃组织中Rh蛋白的阳性反应。结果表明,急性碱度胁迫下,血氨浓度在12 h内快速升高到达峰值,4种Rh基因表达量升高,并于24 h到达峰值;慢性碱胁迫组由于碱度不断升高,血氨浓度呈波动状态,4种Rh基因的表达量均维持在较高表达水平,表明几种Rh蛋白均可能参与血氨浓度调节。胁迫24 h,Rhcg1基因的表达量显著高于其他3种基因。免疫组化结果表明,急性与慢性胁迫组中Rhag、Rhbg和Rhcg在鳃组织中均发现阳性反应,且随碱度升高,阳性反应增强。本研究表明,在碱胁迫环境下,尼罗罗非鱼会增强Rh基因与蛋白表达量参与氨转运过程。     

盐度对珍珠龙胆石斑鱼幼鱼渗透调节与耗氧率的影响

为研究盐度对珍珠龙胆石斑鱼（）渗透调节与耗氧率的影响，设计实验一，将（162.5±12.1）g珍珠龙胆石斑鱼置于不同盐度（6、12、18、24、30）下养殖10 d，测定血清渗透压及Na⁺、Cl^-、K⁺离子浓度。实验结果表明，随盐度的升高血清渗透压及Na⁺、Cl^-、K⁺离子浓度也随着升高，各组[Na⁺]：[Cl^-]比值无显著差异（>0.05）；经回归分析得到血清等渗点渗透压为365.95 mOsm/kg，所对应盐度为12.75。实验二，将（26.4±2.7）g幼鱼置于不同盐度（6、12、18、24、30）下养殖30 d，测定在开始暴露后0 h、3 h、24 h、72 h鳃Na⁺/K⁺-ATPase活性及表达和第30天耗氧率，结果表明鳃Na⁺/K⁺-ATPase活性随盐度的增大呈“U”形变化；鳃Na⁺/K⁺-ATPase α1基因表达量波动较大，在72 h后随盐度增大先降低后增加，变化趋势与酶活性一致；第30天耗氧率随盐度的增加先降低后增加又降低。综上所述，珍珠龙胆石斑鱼幼鱼10 d内能够完全适应6~30盐度急性变化，耗氧率除了受离子渗透调节的影响，还可能与其生活史阶段有关。     

盐碱对大鳞鳃血清渗透压、离子含量及鳃丝Na+/K+-ATP酶活力的影响

通过室内毒理实验,研究了盐度、碱度以及盐碱交互作用对大鳞9E(Barbus capito)血清渗透压、血清离子(Na~+,K~+,CI~-、尿素氮)浓度和鳃丝Na~+/K~+ -ATP酶活力的影响.单因子实验中,随着含盐水平(8 g/L,10g/L、12g/L,14 g/L)的增大,大鳞鳃血清渗透压和血清离子(Na~+,K~+,Cl~-)浓度均显著升高(P<0.05),鳃丝Na~+/K~+-ATP酶活力先升高后降低,受体内外渗透压差的影响最显著(P<0.05).碱度的增大(19.05 mmol/L,30.20 mmol/L,47.86mmol/L,75.86 mmol/L)能引起血清K~+和尿素氮浓度显著升高(P<0.05),但对血清渗透压和鳃丝ATP酶活力无显著影响.在双因子实验中,盐碱交互升高会引起渗透压和血清离子(Na~+,CI~-、尿素氮)显著升高(P<0.05).方差分析结果表明,盐度、碱度及交互作用均对渗透压有显著影响,影响作用最大的是盐度,其次是碱度,交互作用最小.鳃丝Na~+/K~+-ATP酶活性先升高后降低,最高值出现在盐度10 g/L、碱度30.20 mmol/L的正交试验组.从实验结果可得出,大鳞鳃在盐度12g/L以下的水体中能生存,在盐碱共存的环境下,能耐受的上限为盐度10 g/L、碱度30.20mmol.本研究旨在掌握大鳞鳃的生存盐碱度范围,以期为该物种的增养殖生产提供基础依据.     

Sodium dependent ion transporters in trout gills

Biochemical procedures developed to isolate plasma membranes from the branchial epithelium of rainbow trout (Oncorhynchus mykiss) yield membrane fractions that are specifically enriched in the plasma membrane marker enzyme Na⁺/K⁺-ATPase. As the bulk of the branchial Na⁺/K⁺-ATPase is assumed to be confined to the mitochondria-rich chloride cells, such membrane preparations must contain the essence of the enzymatic machinery of the chloride cells. Basal Na⁺ activity in branchial (chloride) cells is around 10 millimolar and, accordingly, we find a K_m for Na⁺ of the Na⁺/K⁺-ATPase of 13 millimolar, indicating that the enzyme may be regulated by changes in cytosolic sodium. The Na⁺-gradient across the serosal plasma membrane created by this pump provides energy for 3Na⁺/Ca²⁺-exchange and bumetanide-sensitive Na⁺/K⁺/2Cl^--cotransport. Here we further postulate the presence of a Na⁺/Cl^--cotransporter, indicated by thiazide-sensitive, bumetanide-insensitive transport of Na⁺ and Cl^-; this cotransporter activity awaits the characterization of its kinetics. The Na⁺/Ca²⁺-exchanger has kinetic characteristics compatible with a regulatory role of cytosolic Na⁺ in the activity of this carrier. Both Na⁺/Ca²⁺-exchange and Ca²⁺-ATPase activity may contribute to transport of Ca²⁺, the former having lower affinity for calcium but a higher capacity than the latter carrier. The Na⁺/K⁺/2Cl^--cotransporter has kinetics that favor a regulatory role for plasma K⁺ in the activity of this carrier. Seawater adaptation leads to increased activity of cotransporter molecules in the plasma membrane fractions (the activity increases relative to that of the Na⁺/K⁺-ATPase) and this may reflect a function in Cl^--extrusion performed by the chloride cells in a seawater environment. A function for the cotransporter in the gills of freshwater fish may be the regulation of cell volume.     

Effects of Na/K and Mg/Ca ratios in saline groundwaters on Na-K-ATPase activity, survival and growth of Marsupenaeus japonicus postlarvae

The effects of the Na⁺/K⁺ and Mg²⁺/Ca²⁺ ratios in saline groundwaters on Na⁺-K⁺-ATPase activity, survival and growth of Marsupenaeus japonicus postlarvae were investigated. The results indicate that the Na⁺-K⁺-ATPase activity, survival rate and weight gain of postlarvae were significantly affected by the Na⁺/K⁺ and Mg²⁺/Ca²⁺ ratios (P < 0.05). The Na⁺-K⁺-ATPase activity of postlarvae, in every treatment, changed corresponding to Na⁺/K⁺ and Mg²⁺/Ca²⁺ ratios, and came to a stable level after 24 h. There was a negative relation between Na⁺-K⁺-ATPase activity and Na⁺/K⁺ ratio, while there was a positive relation between Na⁺-K⁺-ATPase activity and Mg²⁺/Ca²⁺ ratio. Compared with seawater (the Na⁺/K⁺ and Mg²⁺/Ca²⁺ ratios are 27.8 and 4.64 respectively), the Na⁺-K⁺-ATPase activity of the Na⁺/K⁺ ratio 30 treatment showed no significant difference, while the Mg²⁺/Ca²⁺ ratio 4.5 treatment showed distinct difference. The survival rates and weight gain of postlarvae increased markedly when the suitable amount of K⁺ and Ca²⁺ was added to test water, and arrived at their maximum in the Na⁺/K⁺ ratio 20-30 or Mg²⁺/Ca²⁺ ratio 4.5 treatment, having no significant difference compared with normal seawater. Therefore, considering the Na⁺/K⁺, Mg²⁺/Ca²⁺ ratios and the absolute concentration of Mg²⁺, Ca²⁺ in the experimental saline groundwaters applied to Marsupenaeus japonicus farming, it should be modulated to around 30, 4.5 and 1312 mg/l, 291 mg/l, respectively.     

Transepithelial ion exchange in smolting atlantic salmon (Salmo Salar L.)

Whole-body (but predominantly gill) Na⁺ exchange, gill Na⁺/K⁺/ATPase activity and seawater tolerance were examined in juvenile Atlantic salmon during the smolting period. Transepithelial net Na⁺ gain decreased steadily from late February showing a net loss in April and early May, returning to approximate equilibrium in mid-May. This seasonal net loss of Na⁺ to the environment occurred slightly after maximal gill epithelial Na⁺/K⁺/ATPase activity and preceded maximal seawater tolerance. The results are discussed in relation to changes in gill permeability and salt intake via the diet.     

Characterization of transport Na+-ATPases in gills of freshwater tilapia

Branchial plasma membranes from the freshwater cichlid teleostOreochromis mossambicus (tilapia) contain two Na⁺-dependent ATPases: Na⁺/K⁺ ATPase, and an amiloride-sensitive ATPase which is postulated to operate as a Na⁺/H⁺ (–NH₄ ⁺) ATPase. It is suggested that both enzyme activities are located in the basolateral membrane system of the chloride cells. K⁺ has opposing effects on the two enzymes: it stimulates Na⁺/K⁺ ATPase and inhibits Na⁺/H⁺ (–NH₄ ⁺) ATPase activity. Na⁺/H⁺ ATPase appears more sensitive to NH₄ ⁺ at low concentrations than Na⁺/K⁺ ATPase and the stimulatory effect by NH₄ ⁺ ions on the first enzyme could be important in facilitating NH₄ ⁺ excretion by tilapia gills under physiological conditions.In vitro maximum stimulation by NH₄ ⁺ is similar for the two enzymes (200%). In contrast to Na⁺/K⁺ ATPase, Na⁺/H⁺ ATPase activity is inhibited by supra-physiological (>20 mM) concentrations of NH₄ ⁺.     

Mitochondrial enzyme and Na+, K+-ATPase activity,and ion regulation during parr-smolt transformation of Atlantic salmon (Salmon salar)

Atlantic salmon (Salmo salar) exposed to either simulated natural photoperiod (SNP) or continuous light (L24) were used to examine developmental changes in the presence and absence, respectively, of the parrsmolt transformation. Plasma osmolarity and ion concentrations were unaffected by photoperiod treatment. Gill Na⁺, K⁺-ATPase specific activity increased 150% between February and June in SNP fish and was low and unchanged in L24 fish. Kidney Na⁺, K⁺-ATPase specific activity varied within similar, narrow limits in both groups. Citrate synthase of liver, gill and kidney, expressed as specific activity or activity/g total body weight (relative activity), increased 25–60% between March and June in SNP fish. With the exception of kidney relative activity, citrate synthase activity declined to initial (March) levels by August. Liver, gill and kidney cytochrome c oxidase activity of the SNP group underwent similar though less marked changes. Liver, gill and kidney citrate synthase and cytochrome c oxidase activities of the L24 group remained relatively constant between March and August, and where significant differences occurred, they were lower than those of the SNP group. These results indicate that respiratory capacities of the liver, gill and kidney increase in smolls concurrent with preparatory osmoregulatory changes, and subsequently decline. The findings are consistent with a hypothesized transient increase in catabolic activity during the parr-smolt transformation that may be due to the metabolic demands of differentiation.     

Studies on the effect of air exposure on gill Na+/K+-ATPase of the marble goby,Oxyeleotris marmorata,a facultative air-breathing fish

Gill and liver microsomal Na⁺/K⁺-adenosine triphosphatase (ATPase) activities, body weight, and several blood parameters were measured in marble gobies held in freshwater, in air on wet filter paper for 7 days and three days after return to freshwater following 7 days in air. During the 7 days in air, body weight, and blood Na⁺ and K⁺ concentrations remained unchanged. During the same period, however, mean specific activity of the gill ATPase fell 79% while liver ATPase specific activity was unchanged. When these fish were returned to water the specific activity of the gill ATPase returned to values seen in freshwater gobies within 3 days. Several changes were also noted in the characteristics of the ATPase in the fish held in air.     

Osmoregulation in juvenile Siberian sturgeon (Acipenser baerii)

The osmoregulation capabilities of juvenile Siberian sturgeon exposed to three experimental osmolalities (22, 250 and 387 mOsmol kg⁻¹) were studied over a 45-day period. Growth performance, haematological parameters, ion concentrations, gill and spiral valve Na⁺-K⁺-ATPase activities, as well as gill and spiral valve histology, were measured. At the end of the period, the plasma osmolality of fish kept in 250 and 387 mOsmol kg⁻¹ was higher than that of fish kept in 22 mOsmol kg⁻¹. Similar trends were observed in electrolyte concentrations. Spiral valve and gill Na⁺-K⁺-ATPase activity varied with exposure time and environmental salinity. Shortly after being transferred to hyperosmotic media, spiral valve Na⁺-K⁺-ATPase activity fell, while gill Na⁺-K⁺-ATPase activity remained constant. At the end of the experiment, gill Na⁺-K⁺-ATPase activities in fish kept in isosmotic and hyperosmotic media had increased in comparison to those of the control fish. Moreover, spiral valve Na⁺-K⁺-ATPase activities recovered and were similar to those recorded in fish kept in hyposmotic environments. Although some of the morpho-physiological mechanisms were operational in juvenile Siberian sturgeon in their adaptation to hyperosmotic media, fish cannot be considered hyperosmotic regulators as they were unable to maintain their plasma osmolality and electrolyte equilibrium in salinities higher than 250 mOsmol kg⁻¹. This suggests that the culture of juvenile Siberian sturgeon in brackish environments is unlikely to be successful. However, our data indicate that in natural environments, juvenile Siberian sturgeon in migratory populations (Ob and Lena Rivers) would be able to migrate successfully into estuarine brackish grounds with a salinity of up to 9%. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cortisol treatment improves the development of hypoosmoregulatory mechanisms in the euryhaline rainbow trout,Salmo gairdneri

The effect of cortisol on osmoregulatory parameters was studied in rainbow trout, (Salmo gairdneri), kept in freshwater (FW) and/or transferred to seawater (SW). Repeated injections of 20 μg cortisol/g fish stimulated gill and gut Na⁺/K⁺-ATPase activity and reduced plasma Na⁺ and Cl⁻ levels after 2 weeks of treatment in FW-adapted fish. Cortisol doses of 0.05 and 1.0 μg/g were without effect. Repeated injections of 10 μg cortisol/g stimulated gill Na⁺/K⁺-ATPase activity and reduced plasma Na+ and Cl⁻ levels in fish in FW, and significantly improved ion regulation after their transfer to 28SW. Higher doses of cortisol (10 and 20 μg/g) induced hyperglycemia, whereas low doses (0.05 and 1.0 μg/g were without effect or induced hypoglycemia. Plasma glucose levels decreased in cortisol-treated fish transferred to SW, whereas transient hyperglycemia was seen in the control fish.     

Lipid composition and microsomal ATPase activities in gills and kidneys of warm- and cold-acclimated sea bass (Dicentrarchus labrax L.)

The response to cold of gill and kidney membrane lipid composition and microsomal (Na⁺+K⁺)-ATPase, Na⁺-ATPase and Mg²⁺-ATPase activities in reared sea bass (Dicentrarchus labrax L.) was investigated. Fish acclimation was carried out according to the seasonal cycle from August to March. No cold-promoted increase in fatty acid unsaturation was shown in gill and kidney polar lipids and in total lipids of mitochondria and microsomes. In both tissues the (Na⁺+K⁺)-ATPase exhibited positive compensation for cold acclimation whereas the Na⁺-ATPase displayed negative compensation. The Mg²⁺-ATPase showed no compensation in the gills and positive compensation in the kidneys. During cold acclimation the break in the Arrhenius plot of the (Na⁺+K⁺)-ATPase decreased, whereas breaks of both the Na⁺-ATPase and the Mg²⁺-ATPase activities remained unchanged. The results indicate that the sea bass does not adopt membrane unsaturation as a cold-facing strategy. The cold-promoted enhancement of (Na⁺+K⁺)-ATPase activity in osmoregulatory tissues may be advantageous to maintain efficient osmoregulation under thermodynamically unfavourable conditions.     

Quantification of presumptive Na+/H+ antiporters of the erythrocytes of trout and eel

The presumptive Na⁺/H⁺ exchange sites of trout and eel erythrocytes were quantified using amiloride-displaceable 5-(N-methyl-N-[³H]isobutyl)-amiloride (³H-MIA) equilibrium binding to further evaluate the mechanisms of i) hypoxia-mediated modifications in the trout erythrocyte -adrenergic signal transduction system and ii) the marked differences in the catecholamine responsiveness of this system between the trout and eel. MIA was a more potent inhibitor of both trout apparent erythrocyte proton extrusion (IC₅₀ = 20.1 ± 1.1 mol l^–1, N = 6) activity (as evaluated by measuring plasma pH changes after addition of catecholamine in vitro) and specific ³H-MIA binding (IC₅₀ = 257 ± 8.2 nmol l^–1, N = 3) than amiloride, which possessed a proton extrusion IC₅₀ of 26.1 ± 1.6 mol l^–1 (N = 6) and a binding IC₅₀ of 891 ± 113 nmol l^–1 (N = 3). The specific Na⁺ channel blocker phenamil was without effect on adrenergic proton extrusion activity or specific ³H-MIA binding. Trout erythrocytes suspended in Na⁺-free saline and maintained under normoxic conditions possessed 37,675 ± 6,678 (N = 6) amiloride-displaceable ³H-MIA binding sites per cell (B_max, presumptive Na⁺/H⁺ antiporters) with an apparent dissociation constant (K_D) of 244 ± 29 nmol l^–1 (N = 6). Acute hypoxia (PO₂ = 1.2 kPa; 30 min) did not affect the K_D, yet resulted in a 65% increase in the number of presumptive Na⁺/H⁺ antiporters. Normoxic eel erythrocytes, similarly suspended in Na⁺-free saline, possessed only 17,133 ± 3,716 presumptive Na⁺/H⁺ antiporters (N = 6), 45% of that of trout erythrocytes, with a similar K_D (246 ± 41 nmol l^–1, N = 6). These findings suggest that inter- and intra-specific differences in the responsiveness of the teleost erythrocyte -adrenergic signal transduction system can be explained, in part, by differences in the numbers of Na⁺/H⁺ exchange sites.