饥饿对白鲫能量物质消耗的影响

以性成熟的雌性白鲫(Carassius auratus cuvieri)作为试验对象,研究其肌肉、卵巢和肠系膜脂肪等组织能量物质消耗的变化过程和分配关系。分别在饥饿1 d、2 d、4 d、8 d、16 d、24 d、32 d、40 d取样,测定全鱼、肌肉和卵巢的水分、粗蛋白质、粗脂肪和灰分含量。结果显示:全鱼、肌肉的含水量和灰分含量随饥饿时间的延长而升高,而粗蛋白和粗脂肪含量随饥饿时间的延长而减少。在40 d的饥饿中消耗了总蛋白质的7.54%,总粗脂肪的50.94%,其中有17.43%的总蛋白质和27.98%总脂肪在肌肉中被消耗掉。鱼体消耗的蛋白质75%~100%来自肌肉,消耗的脂肪仅有不到10%来自肌肉。脂肪作为主要能量物质被消耗,但也伴随一定量的蛋白质被消耗,其消耗速度不均匀。脂肪在8~16 d出现一个消耗高峰期,蛋白质在24~32 d的消耗量较大。在试验期内其丰满度保持相对稳定,肠系膜脂肪明显减少。饥饿期间卵巢的蛋白质和脂肪含量相对稳定。对照组的水分、粗蛋白质、粗脂肪和灰分含量在试验期内比较稳定。     

Estimating the ideal dietary indispensable amino acid pattern for growth of white sturgeon, Acipenser transmontanus (Richardson)

Juvenile white sturgeon (Acipenser transmontanus, Richardson) were fed or starved for 4 weeks, and their whole body amino acid composition determined at the beginning and the end of the study. The amino acid compositions of muscle, liver, gill and gastrointestinal tract, and the free amino acids in plasma of fed and starved fish were also determined. Very little variation was observed in the amino acid compositions of all the tissues examined after starvation. However, the plasma free amino acids were significantly (P < 0.05) decreased following starvation. On the basis of the amino acid composition and nitrogen retention values of the whole fish, daily indispensable amino acid requirements for protein accretion and obligatory nitrogen losses (mg amino acid 100 g body weighr^?1 day^?1) were estimated. The pattern of indispensable amino acids required for each of these is different, with the pattern for protein accretion containing much higher levels of lysine, but lower levels of tryptophan and sulfur amino acids. Individual amino acid retention rates varied from 33.3% of dietary tryptophan to 80.2% for arginine. The estimated dietary indispensable amino acid pattern for young growing white sturgeon (expressed as a percentage of total indispensable amino acids plus cystine and tyrosine) is: arginine, 14.0; histidine, 6.6; isoleucine, 8.8; leucine, 12.5; lysine, 15.8; methionine (plus cystine), 6.6; phenylalanine (plus tyrosine), 15.5; threonine, 9.7; tryptophan, 0.8; and valine, 9.7.     

The roles of bone and muscle as phosphorus reservoirs during the sexual maturation of female Japanese eels, Anguilla japonica Temminck and Schlegel (Anguilliformes)

We sought to determine the organs that function as phosphorus (P) reservoirs for supplying this mineral to the ovary during the sexual maturation of the Japanese eel, Anguilla japonica (Anguilliformes). The P content of the ovary increased markedly during sexual maturation despite the lack of an extrinsic P supply (e.g., diet). This phenomenon suggests that the P that accumulated in the ovary was derived from other parts of the eel. In addition, the P content of bone dramatically decreased during maturation, as did the total bone weight. This result suggests that resorption of bone occurred during the sexual maturation of the eel. Whereas the P content and mass of muscle markedly decreased during maturation, the Ca and P contents of the skin (which was scaled) did not. The total (whole-fish) P content was constant throughout maturation. Together, our results suggest that almost all of the P lost from bone and muscle is incorporated in the ovary and that bone and muscle function as P reservoirs during the sexual maturation of the Japanese eel. This revised version was published online in August 2006 with corrections to the Cover Date.     

中华绒螯蟹脂肪酸延长酶(ELOVL)基因全长cDNA 的克隆及其表达分析

根据脂肪酸延长酶保守区序列设计引物,采用RT-PCR和RACE技术首次克隆得到中华绒螯蟹(Eriocheir sinensis)脂肪酸延长酶(ELOVL)基因全长(Gen Bank登录号:KR005628)。分析ELOVL序列表明,该基因c DNA全长2089 bp,开放阅读框(ORF)长1065 bp(包含终止密码子),编码的354个氨基酸具有典型的延长酶特征:1个高度保守的氧化还原中心组氨酸簇HVIHH,多个保守区和多个跨膜区(KFTEFLDT、NTFVHIVMYVYY、TNFQMI)。经氨基酸同源性和系统发育树分析,中华绒螯蟹ELOVL预测氨基酸序列与顶切叶蚁(Acromyrmex echinatior)ELOVL氨基酸序列相似性最高,为59%;同时与家蝇(Musca domestica)聚为一支,进而与日本囊对虾(Marsupenaeus japonicus)等聚为一大支。通过实时荧光定量PCR技术研究ELOVL m RNA在中华绒螯蟹不同组织中的表达量,及投喂不同配合饲料后在可食部位组织中的表达情况。结果显示,ELOVL在各组织中均有表达,在肝胰腺和肠道中表达量最高,心脏中最低,其他组织中少量表达。养殖98 d后分析表明,卵巢和肝胰腺组织中ELOVL m RNA在SO饲料组中表达量最高,并且表达水平随着饲料中鱼油(富含n-3 PUFA)比例减少、豆油(缺少n-3 PUFA)比例增加而显著升高(P0.05);精巢组织中ELOVL m KNA表达量在SO饲料组中最高;肌肉组织中ELOVL m KNA表达水平较低,且各饲料组间表达量差异不显著(P0.05)。以上结果表明,中华绒螯蟹存在ELOVL基因,并且ELOVL的表达受饲料脂肪水平的影响,这为探究中华绒螯蟹自身合成HUFA途径及调节机制奠定了基础。     

The changes in the biochemical compositions and enzymatic activities of rotifer (<Emphasis Type="Italic">Brachionus plicatilis</Emphasis>, Müller) and <Emphasis Type="Italic">Artemia</Emphasis> during the enrichment and starvation periods

The changes in the biochemical compositions and enzymatic activities of rotifer (Brachionus plicatilis) and Artemia, enriched and stored at 4°C temperature, were determined. The total starvation period was 16 h and samples were taken at the end of the 8th and 16th hours. In present study, the rotifer and nauplii catabolized a large proportion of the protein during the enrichment period. Lipid contents of both live preys increased during the enrichment period and decreased in nauplii and metanauplii throughout the starvation period but lipid content of the rotifer remained relatively constant during the starvation period. The changes observed in the amino acid compositions of Artemia and the rotifer were statistically significant (P < 0.05). The conspicuous decline the essential amino acid (EAA) and nonessential amino acid (NEAA) content of the rotifer was observed during the enrichment period. However, the essential amino acid (EAA) and nonessential amino acid (NEAA) contents of Artemia nauplii increased during the enrichment period. The unenriched and enriched rotifers contained more monounsaturated fatty acid (MUFAs) than polyunsaturated fatty acid (PUFAs) and saturated fatty acids (SFA). However, Artemia contained more PUFAs than MUFAs and SFA during the experimental period. A sharp increase in the amounts of docosahexaenoic acid (DHA) during the enrichment of the rotifer and Artemia nauplii was observed. However, the amount of DHA throughout the starvation period decreased in Artemia metanauplii but not in Artemia nauplii. Significant differences in tryptic, leucine aminopeptidase N (LAP), and alkaline phosphatase (AP) enzyme activities of Artemia and rotifer were observed during the enrichment and starvation period (P < 0.05). The digestive enzymes derived from live food to fish larvae provided the highest contribution at the end of the enrichment period. In conclusion, the results of the study provide important contributions to determine the most suitable live food offering time for marine fish larvae. Rotifer should be offered to fish larvae at the end of the enrichment period, Artemia nauplii just after hatching and before being stored at 4°C, and Artemia metanauplii at the end of the enrichment and throughout the starvation period.     

中华绒螯蟹Akt基因的cDNA克隆、序列分析及表达特征

为研究Akt基因在中华绒螯蟹蜕壳前后肌肉生长过程中的功能,应用RACE技术克隆得到编码中华绒螯蟹Akt(命名为Es Akt)的全长为2 200 bp的c DNA序列,包括159 bp的5′非翻译区(5′-UTR)、496 bp的3′非翻译区(3′-UTR)和长度为1545 bp编码514个氨基酸的编码序列。蛋白质结构域分析显示Es Akt含有丝氨酸/苏氨酸蛋白激酶家族的3个特征性保守结构域。多序列比对和系统发育分析显示,Es Akt与中国明对虾、凡纳滨对虾的Akt序列一致性都为0.889,在系统发育树中节肢动物Akt形成一个分支。应用荧光定量RTPCR技术分析Es Akt在性成熟中华绒螯蟹各组织及幼体不同蜕壳时期不同部位肌肉组织中转录水平上表达量的变化。结果显示,Es Akt在性成熟个体的肝胰腺、眼柄、表皮、卵巢、精巢、心脏、螯足、鳃、三角膜等组织中均有表达,其中卵巢、眼柄和精巢中表达量较高,肝胰腺中表达量最低。在幼体不同蜕壳时期不同部位的肌肉中,Es Akt表达量变化不同,步行足肌肉组织中Es Akt m RNA无显著的统计学差异。腹部肌肉组织中Es Akt m RNA水平在蜕壳前晚期D3~D4期表达量显著高于蜕壳后A~B期和蜕皮间期C期。螯足肌肉在蜕壳前晚期D3~D4期急剧下调,蜕壳后A~B期开始表达量显著升高,直至蜕皮间期C期。研究表明,Es Akt在中华绒螯蟹蜕壳过程中不同部位肌肉组织中的表达量变化与蜕壳周期密切相关,说明Es Akt参与中华绒螯蟹蜕壳诱导的肌肉萎缩、生长及重建过程。     

Effect of fish feed processing conditions on digestive protease activities, free amino acid pools, feed conversion efficiency and growth in Atlantic salmon (Salmo salar L.)

The responses of the digestive proteases trypsin and chymotrypsin and protein metabolism to differences in feed protein quality were investigated in Atlantic salmon (Salmo salar L.). Two sets of experimental feeds were produced. Each set of high and low quality feeds was provided to either 150 g or 2 kg salmon. Protein in the high quality feeds had significantly higher percentages of free (reactive) sulphydryl (SH) groups than the corresponding feeds based on low quality meals. After 90 days feeding, groups given high and low quality feeds did not differ in their specific growth rates (SGR) in either experiment. However, feed conversion efficiency (FCE) was significantly different between the high and low quality feed groups in 2 kg salmon, where the difference between the high and low feed protein qualities was larger, 10% versus 4% SH/[SH + (S–S)] in 150 g salmon. Higher FCE was preceded by significantly higher trypsin and chymotrypsin specific activities on day 60. SGR, in general, changed after the first month and was stable during the last 2 months in both experiments. Concurrently, both trypsin (T) and chymotrypsin (C) decreased with an increased activity ratio of trypsin to chymotrypsin (T/C ratio), and resulting in significantly lower T/C ratio on day 90 in salmon feeding on high quality feeds in both sizes of fish. Differences in FCE were associated with significant differences in levels of total free amino acids (TFAA) in the plasma and the white muscle, as well as in the ratio of essential to non‐essential free amino acids (EAA/NEAA ratio), free hydroxyproline, and RNA in the white muscle. Interestingly, after 3 days starvation (day 93), 5–7 h postprandial EAA/NEAA ratio in the plasma was significantly lower in the high quality diet groups in both experiments. Trypsin specific activity inversely correlated with muscle TFAA levels in 2 kg salmon, concurrent with higher muscle levels of RNA, lower free hydroxyproline and higher FCE in fish fed higher quality diets.     

东太平洋赤道海域茎柔鱼体征生长及生殖投入

现生头足类的性腺发育滞后于个体生长,研究其生殖投入方式对于掌握它们的生长发育过程有着重要的意义。为此,本实验利用形态学和残差指标分析方法,对东太平洋赤道海域茎柔鱼的体征生长及其生殖投入进行了分析。结果显示,雌、雄个体均为匀速生长,雌性个体性腺成熟的最小胴长和最小体质量分别为270 mm和766 g,雄性个体性腺成熟的最小胴长和最小体质量分别为270 mm和479 g。雌、雄个体的空胃率低,摄食等级以1～2级为主;随着月份推移,个体的体征质量逐步趋好,在性腺发育阶段最差。雌性成熟个体的性腺指数平均值为2.26%±1.33%,缠卵腺指数平均值为34.34%±18.62%;雄性成熟个体的性腺指数平均值为1.31%±0.35%。雌、雄个体二者的体征指标与生殖投入指标均呈显著的正相关关系;随着性腺发育,体征指标和生殖投入指标呈下降趋势;随着胃饱满度等级增加,体征指标呈下降趋势,生殖投入指标则呈上升趋势。研究表明,茎柔鱼在性腺发育过程中持续摄食生长,生殖投入倾向于外源性,肌肉组织存储能量部分转化以满足生殖发育所需。     

Digestive efficiency,free amino acid pools and quality of growth performance in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.) affected by light regimes and vaccine types

This study comprised the results of three different seawater trials using unique combination of techniques to study protease digestive efficiency and growth performance quality to illustrate the effects of light regimes and vaccine types in Atlantic salmon (Salmo salar L.). Fish with higher growth had higher trypsin (T) and chymotrypsin (C) specific activities with higher T/C ratio or slope T/C ratio [calculated from the regression between trypsin (y) and chymotrypsin (x) specific activities] in the pyloric caeca. The T/C ratios indicated fish growth rates over a period of 1–2 months, while the slope T/C ratios indicated fish growth rates at sampling. Adaptation period for adjustment to the new environment of continuous light was 70 days, indicated by the differences in trypsin specific activities and the crossing of slope T/C ratio regressions following with the changes in growth rate directions between the control and the treated group. Vaccine types affected fish vertebral growth, and additional continuous light enhanced the impact of vaccines on fish growth during springtime, indicated by differences in slope T/C ratios. Continuous light stimulated fish growth during winter to spring, when the natural day length was short, without significantly changing white muscle and oocyte qualities in the fish of about 500 g, except for significantly increased white muscle RNA concentration. Continuous light also reduced fish growth rate later during summer, when the natural day length was long, by precedently decreasing the T/C ratio in late spring. Interestingly, plasma levels of free lysine related to tryptic digestion were correlated with trypsin specific activity levels. Continuous light caused higher levels of most free amino acids (FAA) involved in nitrogen metabolism, higher incorporation of essential FAA for protein synthesis, and higher protein turnover rate (free hydroxyproline levels) in both plasma and white muscle. However, continuous light did not affect higher protein content, intracellular buffering capacity and RNA levels in the white muscle of the fish of about 1 kg, probably due to limitation of FAA available for protein synthesis. It is therefore suggested that enhancing fish growth by continuous light stimulation should be accompanied by increasing availability or content of dietary protein (and probably minerals), which in turn would improve the quality of fish growth performance through increasing fillet protein concentration, strengthening vertebral growth, and delaying oocyte development.     

饥饿对大黄鱼幼鱼肌肉中氨基酸和脂肪酸组成的影响

为了研究饥饿对大黄鱼(Larimichthys crocea)幼鱼肌肉中氨基酸和脂肪酸含量的影响,取大黄鱼幼鱼540尾,体重均值为(40.80±3.40)g,分组进行为期0 d(S0)、4 d(S4)、8 d(S8)、12 d(S12)、16 d(S16)、20 d(S20)的饥饿处理,测定背部肌肉的氨基酸和脂肪酸含量。结果显示,测定的大黄鱼背肌的16种氨基酸中,蛋氨酸含量在不同饥饿处理时间之间差异显著(P0.05),饥饿持续8 d其含量达到最小(0.48%±0.12%),16 d时达到最高(1.62%±0.23%);其余15种氨基酸含量的差异均不显著(P0.05),但均表现出随着饥饿时间延长先上升后下降的趋势,在16 d时达到最高,20 d时明显降低。非必需氨基酸、必需氨基酸、呈味氨基酸、鲜味氨基酸及氨基酸总量的变化趋势与上述15种氨基酸一致。在不同饥饿处理组的大黄鱼背肌中脂肪酸含量差异显著(P0.05)。其中,饱和脂肪酸(SFA)含量随着饥饿时间延长呈先升高后下降的趋势,在S8组达到最大,为30.90%±0.28%;单不饱和脂肪酸(MUFA)含量在不同处理组之间差异不显著(P0.05),但所有处理组均稍大于对照组(S0);多不饱和脂肪酸(PUFA)含量则呈先下降后升高的趋势,在S12组时达到最低,其值为31.87%±0.65%。由上可知,通过适当的饥饿处理,可以改变肌肉中氨基酸和脂肪酸含量,从而较好地改善大黄鱼的肉质风味。     

育肥时间对三疣梭子蟹卵巢发育和营养品质的影响

采用育肥养殖实验和生化分析技术,研究了育肥时间对三疣梭子蟹雌体卵巢发育和营养品质的影响。结果发现,(1)经90 d育肥的雌体卵巢指数显著上升,出肉率和总可食率呈先上升后下降趋势,育肥30~60 d的总可食率达42%以上。(2)育肥时间对肌肉中常规营养成分含量无显著影响,育肥60 d卵巢和肝胰腺中的蛋白质、脂肪和碳水化合物含量最高,水分含量最低;育肥使肝胰腺和卵巢中胆固醇百分含量呈显著下降趋势,育肥后期卵巢和肝胰腺中的磷脂含量显著上升;就不同组织而言,肝胰腺中的甘油三酯含量最高。(3)育肥使卵巢和肝胰腺中的单不饱和脂肪酸呈上升趋势,而多不饱和脂肪酸呈先下降后上升趋势,肌肉中脂肪酸组成无显著变化;整体上,肌肉的脂肪酸营养价值优于肝胰腺和卵巢。(4)育肥早期卵巢中氨基酸含量显著上升,整个育肥过程中肌肉中的甜味氨基酸含量呈显著上升趋势,育肥60 d后卵巢和肌肉中所有必需氨基酸分值均大于100,无限制性氨基酸。研究表明,生殖蜕壳后的三疣梭子蟹雌体适宜育肥时间为60 d,育肥不仅提高了总可食率,且优化了其营养价值。     

Changes in tissue and plasma free amino acid concentrations after feeding in Atlantic cod

Atlantic cod, Gadus morhua, were maintained on a diet of sandeel and after a 6-day fast were refed a single meal. Concentrations of free amino acids (AAs) were measured in hepatic portal and cardiac blood as well as in the stomach and white muscle at intervals of 6h up to 24h post-feeding. The appearance of both essential and non-essential AAs in the hepatic portal blood was significantly correlated, up to 12h after feeding, to their abundance in the diet. There was a significant decline in total AA concentration in cardiac blood after 6h, followed by a significant increase at 12h. No significant changes in total AA concentration were observed in the other tissues, although mean concentration increased at 12 or 18h. At a more detailed level, the post-prandial changes in concentration of some essential AAs were consistent with their having a role in the stimulation of protein synthesis after feeding.     

Oxidative stress parameters during starvation and refeeding periods in Adriatic sturgeon (Acipenser naccarii) and rainbow trout (Oncorhynchus mykiss)

This work analyses the changes in the redox balance in two fish species: Adriatic sturgeon ( Acipenser naccarii ) and rainbow trout ( Oncorhynchus mykiss ) during starvation and refeeding period. The starvation period raised the lipid peroxidation (thiobarbituric-acid-reacting substances) levels in liver and blood, while a decline occurred in the antioxidant enzymatic activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and glutathione reductase (GR) in both fish species. In liver, after the refeeding period, SOD activity recovered in both species, whereas CAT activity recovered only in trout. Furthermore, in both tissues of the two species, the lipid peroxidation levels remained high after 2 months of refeeding. In white muscle and heart, the lipid peroxidation levels indicate that these tissues did not undergo oxidative stress during the 72-day period. During starvation, in the muscle of both fish the fall in the lipid peroxidation level coincided with a rise in CAT, GPX and GR. The refeeding period in this tissue raised the lipid peroxidation level, and the enzymatic activities reached the values of the first point of starvation. In heart, no oxidative damage was detected during starvation in either species. The CAT and SOD activities increased during the starvation period only in trout.     

凡纳滨对虾精氨酸激酶的多克隆抗体制备及组织特异性表达分析

精氨酸激酶（arginine kinase,AK）在调节无脊椎动物磷酸精氨酸与ATP之间能量平衡的过程中具有重要作用。在前期工作基础上,设计特异引物,以凡纳滨对虾肌肉组织cDNA为模版,克隆得到了1 071 bp的凡纳滨对虾精氨酸激酶的完整开放阅读框;将它克隆到原核表达载体pGEX-4T-2上,转化BL21 (DE3) 菌株,经诱导后表达出GST-AK融合蛋白。以纯化的融合蛋白作为抗原免疫小鼠,制备多克隆抗血清,经检测该抗原与抗体识别良好。利用得到的抗体对凡纳滨对虾AK在蛋白质水平上的特异性表达进行分析发现,精氨酸激酶在对虾的肌肉、心脏、神经、血细胞和胃组织中具有高的表达量,而在眼柄、肝胰腺、鳃和皮肤组织中表达量较低。为进一步研究精氨酸激酶在对虾体内的功能奠定了基础。     

饥饿对曼氏无针乌贼肝脏与卵巢脂肪酸组成的影响

在曼氏无针乌贼（Sepiella maindroni）性腺成熟阶段（4～5月）对其进行饥饿处理,以空腹处理组饥饿0d（禁食1d）、饥饿12d（禁食13d）和饥饿24d（禁食25d）为取样时间取其肝脏和卵巢组织进行脂肪酸组成及质量分数测定。结果表明,随着饥饿时问的延长,乌贼体质量下降,肝脏和卵巢萎缩,肝脏中的饱和脂肪酸（SFA）和单不饱和脂肪酸（MUFA）较空腹处理组有显著下降（P〈0．05）,而多不饱和脂肪酸（PUFA）和高不饱和脂肪酸（HUFA）均有显著提高（P〈0．05）。与肝脏相比,乌贼卵巢中的各脂肪酸组成在饥饿12d变化较小,饥饿24d时∑SFA较空腹处理组有极显著下降（P〈0．01）,而∑MUFA、∑PUFA＋∑HUFA和∑n-6 PU—FA显著高于空腹处理组（P〈0．05）。     

Metabolic adjustments of Dentex dentex to prolonged starvation and refeeding

The particular metabolic strategies of the common dentex (Dentex dentex) to face a period of prolonged starvation and subsequent refeeding were assessed. Plasma metabolites, endogenous reserves, and the activity of key enzymes of intermediary metabolism in liver, white muscle, and heart were evaluated. Plasma glucose, total lipid, triglycerides, total-, HDL- and LDL-cholesterol, and protein levels, liver, and white muscle glycogen, and perivisceral, and muscle fat were significantly reduced by starvation, whereas liver lipid content was surprisingly increased. Those enzymes involved in phosphorylation and oxidation of glucose and lipid synthesis, as well as alanine aminotransferase activity, were significantly depressed in liver of starved fish. The increase in β-hydroxyacyl-CoA dehydrogenase (HOAD) indicated an enhanced fatty acid oxidation during starvation. Part of the acetyl-CoA generated by β-oxidation was oxidized in the hepatic Krebs cycle, as reflected the increased citrate synthase (CS) activity. The oxaloacetate required for the reaction catalized by CS activity would be supplied by aspartate aminotransferase (ASAT) activity whose activity was also enhanced. Glutamate dehydrogenase also increased to deaminate the glutamate produced by transaminases, especially by the increased ASAT activity. Liver gluconeogenesis of starved fish was maintained at the same rate that in controls, with glycerol playing an important role as glucogenic substrate. The increased hepatic β-hydroxybutyrate dehydrogenase (β-OHBDH) activity indicates that part of the acetyl-CoA arriving from β-oxidation was being diverted for ketone bodies production with dentex liver playing an important role in providing ketone bodies as fuels for other tissues under such circumstances. Most enzyme activities in white muscle of starved dentex were significantly depressed. In heart, starvation induced an important inhibition of those enzymes involved in glucose and protein metabolism, whereas CS, HOAD, and β-OHBDH activities were maintained at control levels. Although several biomarkers assayed returned to control values after refeeding, many others did not, which indicate that after 3 weeks of refeeding, pre-starved dentex is still experiencing a transient period of metabolic adjustments directed toward the restoration of body mass.     

Effect of feeding on the tissue free amino acid concentrations in rainbow trout (Oncorhynchus mykiss Walbaum)

This study investigates whether tissue free amino acid (FAA) pools in rainbow trout,Oncorhynchus mykiss (Walbaum), are altered following feeding and the relationships between the amount of food consumed and the FAA pool size. Trout were starved for 7 days to provide baseline data and then refed on day 8. Individual food intake was measured by radiography and the consumption of amino acids (AA) calculated from dietary protein consumption. Total FAA concentrations in the stomach, liver and white muscle were little changed at various times after the meal and this pattern was repeated for the majority of individual FAA. Overall, the most notable change was a reduction in essential FAA concentrations (principally in valine, leucine and isoleucine) in the white muscle following feeding. However, in the caeca total FAA, total essential FAA and a number of individual FAA were significantly elevated at 4, 9 and 15h following feeding. There were few significant correlations between dietary amino acid consumption and total tissue FAA and essential FAA concentration in the stomach, caecum and white muscle; correlations were stronger in the liver. In order to explain the relative constancy of total FAA concentrations in the tissues following food intake (which represents over 100% of the total FAA pool) a model is presented that quantifies the AA flux through the free pools and considers the role of protein turnover in regulating FAA pool size.     

禁食状态下凡纳滨对虾游离氨基酸的降解代谢

在禁食胁迫下,对在盐度为0(S0)和30(S30)水体中的凡纳滨对虾肌肉游离氨基酸(FAA)的浓度变化进行测定.结果表明,禁食15d后,两种盐度中的凡纳滨对虾肌肉游离氨基酸总量均有所升高,但其中游离的牛磺酸(Tau)、苏氨酸(Thr)、脯氨酸(Pro)、丙氨酸(Ala)、半胱氨酸(Cys)、缬氨酸(Val)、蛋氨酸(Met)、异亮氨酸(Ile)、亮氨酸(Leu)、苯丙氨酸(Phe)、赖氨酸(Lys)、组氨酸(His)的浓度却有不同程度的降低,脯氨酸(Pro)在禁食15d后的浓度降为0,牛磺酸(Tau)、苏氨酸(Thr)、丙氨酸(Ala)、半胱氨酸(Cys)、苯丙氨酸(Phe)、赖氨酸(Lys)的降幅也比较显著(P<0.05);但甘氨酸(Gly)、天门冬氨酸(Asp)、酪氨酸(Tyr)、精氨酸(Arg)等主要游离氨基酸的浓度却显著升高,从而使游离氨基酸总浓度升高.实验表明,凡纳滨对虾肌肉里游离氨基酸浓度在禁食胁迫下降解代谢强度不一,饲料中添加脯氨酸(Pro)、精氨酸(Arg)和赖氨酸(Lys)等游离氨基酸后,可能会对凡纳滨对虾的生长起到促进作用.     

斑节对虾谷氨酸脱氢酶基因的克隆及氨氮胁迫对其时空表达的影响

采用cDNA末端快速扩增技术(rapid amplification of cDNA ends, RACE)获得了斑节对虾(Penaeus monodon)谷氨酸脱氢酶基因(PmGDH)的cDNA序列。该序列全长2386 bp,开放阅读框(ORF)为1677 bp,3?非编码区(UTR)为688 bp,包括含有27个碱基的poly(A)尾,5?非编码区(UTR)为21 bp。ORF可编码558个氨基酸,预测分子量为61.837 kD,理论等电点为6.57。序列含有ELFV dehydrog N与NAD bind 1 Glu DH两个保守结构域,37个磷酸化位点,3个糖基化位点。通过同源性、相似性以及系统进化树分析,斑节对虾的GDH基因与凡纳滨对虾(Litopenaeus vannamei)GDH基因的同源性和相似性最高,并与其聚为一支。采用荧光定量的方法研究了PmGDH基因在斑节对虾不同组织、氨氮胁迫过程中差异表达情况。结果表明, PmGDH的mRNA在各组织中都有表达,其中,在肌肉组织中表达量最高,其次为眼柄神经,在血淋巴与肠组织中表达量最低。96 h氨氮胁迫后,荧光定量PCR分析结果表明PmGDH在肝胰腺和鳃组织中的表达与对照组相比都具有显著差异(P<0.05),都具有不同程度的表达上调。表明PmGDH在氨氮代谢方面具有重要的作用,参与了斑节对虾机体的急性氨氮胁迫应答反应。     

三疣梭子蟹PtDNMT1基因的克隆及其 在低盐适应中的表达分析

甲基转移酶是维持基因组甲基化状态的重要基因,本研究利用SMART-RACE技术克隆了三疣梭子蟹(Portunus trituberculatus)甲基转移酶基因(PtDNMT1)。PtDNMT1基因cDNA序列全长5919 bp,包括4832 bp的开放阅读框,编码1610个氨基酸,预测分子量为148.15 kDa,理论等电点为4.68。结构预测发现,PtDNMT1有2个特殊的结构域,分别是锌指结构域(zf-CXXC)和甲基转移酶家族特有的Dcm结构域。进化树分析显示,PtDNMT1基因与昆虫类的DNMT基因聚为一支。组织表达分析发现,PtDNMT1基因在肝胰腺、鳃、卵巢、肌肉、胃、心脏、血液中均有表达,其中在肝胰腺中表达最高,卵巢和鳃次之。进一步研究了低盐胁迫后PtDNMT1基因在鳃、肝胰腺和肌肉组织中的表达变化规律为胁迫6 h时鳃组织中PtDNMT1基因的表达即达到峰值(5.3倍),并一直持续到12 h (4倍),随后逐步下降,在72 h时仍显著高于对照组(2.3倍);PtDNMT1基因在肝胰腺中的表达规律类似于鳃,然而其达到峰值的时间稍晚于鳃(24 h),且上调倍数高于鳃(8倍);低盐胁迫后PtDNMT1基因在肌肉中的表达最初呈现下调趋势,之后(24 h)上调表达至峰值(2.2倍),且一直上调表达至72 h。本研究首次克隆了PtDNMT1基因,根据其在各组织中的表达分布特征以及盐度胁迫后的表达变化情况,推测DNA甲基化在三疣梭子蟹低盐适应中发挥了重要作用。