鲤鳞被SLC25A5基因的克隆和表达分析

本研究利用差异表达技术(Genefishing)在德国镜鲤和建鲤两个皮肤中表达差异,分离和克隆了鲤鱼鳞被相关的基因,溶质载体蛋白基因SLC(Solute Carrier Protein)大家族中SLC25基因小家族,通过克隆测序的序列在NCBI中比对结果分析,和SLC25家族中的SLC25A5基因结构非常相似,同源性达到9 9%。     

鲤鱼微卫星分子标记的筛选

微卫星 (ｍｉｃｒｏｓａｔｅｌｌｉｔｅ)是近十几年来发展起来的一种新的分子标记 ,它是指以少数几个核苷酸 ( 1～ 6个 )为单位多次重复的简单序列 ,以双核苷酸重复最为常见 ,而其中又以 (ＣＡ/ＧＴ) ｎ 居多。由于微卫星在真核生物基因组中是随机分布的 ,而作为分子遗传标记又有着非常高的多态性和共显性 ,因此在构建遗传连锁图谱时备受青睐 (Ｂｒｏｏｋｅｔａｌ ,1994 )。目前 ,在人类和多种动物中已经构建了以微卫星为主的遗传连锁图谱。但在鲤鱼等水产动物的遗传连锁图谱中 ,微卫星分子标记还较少 (孙效文和梁利群 ,2 0 0 0 )。为了摸索…     

转反义sGnRH基因败育鲤鱼的性成熟诱导

采用反义转基因技术已成功获得整合CAsGnRHpc-antisense基因的转基因鲤鱼,简称AS-sGnRH F0鲤鱼.通过挤压1年龄AS-sGnRH F0鲤鱼的腹部,发现该群体中有精液/无精液个体比例为33/69,与对照组鲤鱼中有精液/无精液个体比例接近1：1相比存在显著差异.通过注射鲤鱼垂体抽提物诱导无精液AS-sGnRH F0鲤鱼恢复繁殖功能,获得3尾育性被恢复的AS-sGnRH F0鲤鱼.进一步的解剖观察发现,在无精液AS-sGnRH F0鲤鱼群体中,完全无性腺的AS-sGnRH F0鲤鱼比例约31.3%.根据直观的挤精液和解剖学观察,推测在AS-sGnRH F0鲤鱼群体中,雄鱼：雌鱼：＂脂肪型＂性腺个体的比例大约为35：45：20%.受精卵发育12 h后,统计A、B、C系AS-sGnRH F0鲤鱼精子的受精率分别为73.94、59.45和64.05%.从A、B、C系杂交后代F1分别随机取样100尾,PCR检测的阳性率分别为0、78.1和87.8%.研究结果表明通过补充外源激素可以使促性腺激素分泌不足而不育的AS-sGnRH F0鲤鱼恢复具有产生成熟配子的能力.     

Rheological Characteristics and Microstructure of Common Carp (Cyprinus carpio) Surimi and Kamaboko Gel

Common carp is cheap and prolific in Australian waters and is regarded as an aquatic environmental pest. In order to add value to this fish species, surimi and kamaboko was prepared from common carp and its rheological and microstructure characteristics were compared with those produced from Alaska pollock and threadfin bream. Temperature sweep tests were run under 100-Pa stress and 0.1-Hz frequency, obtained from linear viscoelastic ranges of all tested fish gels. Storage modulus (G′) thermographs of all samples consisted of two positive peaks and a plateau zone in between. The sol-gel transition was completed at about 53 and 61 °C for these Alaska pollock and threadfin bream gels, respectively, whereas it was recorded at about 69 °C for common carp gel. At these temperatures, G′ of Alaska pollock gel was recorded at 330 kPa, which was 71% and 88% greater than that from threadfin bream and common carp gels, respectively. Furthermore, Alaska pollock and threadfin bream gels had greater gel strength than the gel prepared from common carp surimi. Textural quality could be associated with cross-linking in the gel network. From scanning electron microscopy studies, the microstructure of Alaska pollock gel matrix had about 15,450 polygonal structures per square millimeter with an average area of about 9 μm². For threadfin bream and common carp gels, the polygonal structures were larger and 12% and 39% fewer, respectively, than those of Alaska pollock gel. However, these results cannot be only attributed to the species variation among tested fish as some other factors such as harvest location, physiological state, handling and processing method, etc. were not considered in this study.     

Toxic Effects of Cobalt Chloride on Hematological Factors of Common Carp (Cyprinus carpio)

In this study, we investigate the toxic effects of cobalt chloride on some hematological factors of the carp Cyprinus carpio, such as white blood cell count, red blood cell count, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration. At first, LC₅₀ of cobalt in C. carpio was measured during 96 h after exposure. Also, physicochemical parameters of water including pH, dissolved oxygen, viscosity, temperature, and conductivity were monitored, continuously. The results showed that LC₅₀ values of cobalt in C. carpio were 327 and 328 mg/L in two replicates, respectively. Then, the changes in some hematological factors in the five treatment groups placed under concentration of 100, 200, 300, 400, and 500 mg/L cobalt were compared with the control group. Based on hematological tests conducted in this research, exposure of carp to 500- and 300-mg/L concentrations of cobalt in 48 h showed significant difference (p?<?0.05) in white blood cell count. The concentration of 500 mg/L cobalt in 24 h showed a significant difference in the amount of hemoglobin, number of red blood cells, and hematocrit level as compared with the control group. The concentration of 100 mg/L cobalt in 48 h did not show a significant difference in comparison with the control group (p?>?0.05). Also, the concentration of 500 mg/L cobalt in 24 h showed a significant difference in the amount of mean corpuscular volume and mean corpuscular hemoglobin as compared with the control group and other treatments. Also, the percentage of mean corpuscular hemoglobin concentration in a concentration of 200 mg/L cobalt in 24 h showed a significant difference as compared with the control group and other treatments.     

Genetic Linkage Mapping and Analysis of Muscle Fiber-Related QTLs in Common Carp (Cyprinus carpio L.)

A genetic linkage map of common carp (Cyprinus carpio L.) was constructed using Type I and Type II microsatellite markers and a pseudo-testcross mapping strategy. The microsatellite markers were isolated from microsatellite-enriched genomic libraries and tested for their segregation in a full-sib mapping panel containing 92 individuals. A total of 161 microsatellite loci were mapped into 54 linkage groups. The total lengths of the female, male and consensus maps were 2,000, 946, and 1,852?cM, with an average marker spacing of approximately 13, 7, and 11?cM, respectively. Muscle fiber-related traits, including muscle fiber cross-section area and muscle fiber density, were mapped to the genetic map. Three QTLs for muscle fiber cross-section area and two QTLs for muscle fiber density were identified when considering both significant and suggestive QTL effects. The QTLs with largest effects for muscle fiber cross-section area and muscle fiber density were 21.9% and 18.9%, and they were located in LG3, respectively.     

鲤幼鱼标准代谢率的个体差异与运动性能和摄食代谢的关系

为考察鲤科鱼类种内个体标准代谢率的差异及其与运动性能和摄食性能的内在关联,本研究以我国广泛分布的鲤(Cyprinids cardio)幼鱼[体重(4.79±0.08)g,n=36]为实验对象,在(25.0±1.0)℃下分别测量实验鱼的标准代谢率(SMR),随后测定单尾鱼的特殊动力作用(SDA)、自发运动、临界游泳速度以及活跃代谢率(MO2active)。实验鱼标准代谢率(SMR)的变幅为76.7~317.6 mg/(kg·h),其变异系数(CV)达24.4%;实验鱼在10 min内的尾鳍摆动次数(P0.05)和摄食代谢峰值(P0.05)均与标准代谢率(SMR)呈正相关;活跃代谢率(MO2active)(P0.05)与摄食代谢峰值以及活跃代谢范围与摄食代谢范围(P0.05)均呈正相关。然而,鲤幼鱼的标准代谢率(SMR)与相对临界游泳速度、活跃代谢率(MO2active)、特殊动力作用(SDA)时间和特殊动力作用(SDA)总量均不相关(所有P0.05)。研究表明,较高标准代谢率(SMR)的鲤幼鱼个体表现较高的活跃性和较强的摄食代谢能力,可能有助于其更易发现食物、逃避天敌以及加快食物处理。     

Genome-Wide Identification,Characterization and Phylogenetic Analysis of ATP-Binding Cassette (ABC) Transporter Genes in Common Carp (Cyprinus carpio)

The ATP-binding cassette (ABC) gene family is considered to be one of the largest gene families in all forms of prokaryotic and eukaryotic life. Although the ABC transporter genes have been annotated in some species, detailed information about the ABC superfamily and the evolutionary characterization of ABC genes in common carp (Cyprinus carpio) are still unclear. In this research, we identified 61 ABC transporter genes in the common carp genome. Phylogenetic analysis revealed that they could be classified into seven subfamilies, namely 11 ABCAs, six ABCBs, 19 ABCCs, eight ABCDs, two ABCEs, four ABCFs, and 11 ABCGs. Comparative analysis of the ABC genes in seven vertebrate species including common carp, showed that at least 10 common carp genes were retained from the third round of whole genome duplication, while 12 duplicated ABC genes may have come from the fourth round of whole genome duplication. Gene losses were also observed for 14 ABC genes. Expression profiles of the 61 ABC genes in six common carp tissues (brain, heart, spleen, kidney, intestine, and gill) revealed extensive functional divergence among the ABC genes. Different copies of some genes had tissue-specific expression patterns, which may indicate some gene function specialization. This study provides essential genomic resources for future studies in common carp.     

Chronic Exposure to Tributyltin Induces Brain Functional Damage in Juvenile Common Carp (Cyprinus carpio)

The aim of the present study was to investigate the effect of Tributyltin (TBT) on brain function and neurotoxicity of freshwater teleost. The effects of long-term exposure to TBT on antioxidant related indices (MDA, malondialdehyde; SOD, superoxide dismutase; CAT, catalase; GR, glutathione reductase; GPx, glutathione peroxidase), Na⁺-K⁺-ATPase and neurological parameters (AChE, acetylcholinesterase; MAO, monoamine oxidase; NO, nitric oxide) in the brain of common carp were evaluated. Fish were exposed to sublethal concentrations of TBT (75 ng/L, 0.75 μg/L and 7.5 μg/L) for 15, 30, and 60 days. Based on the results, a low level and short-term TBT-induced stress could not induce the notable responses of the fish brain, but long-term exposure (more than 15 days) to TBT could lead to obvious physiological-biochemical responses (based on the measured parameters). The results also strongly indicated that neurotoxicity of TBT to fish. Thus, the measured physiological responses in fish brain could provide useful information to better understand the mechanisms of TBT-induced bio-toxicity.     

微卫星DNA标记分析野生鲤鱼群体的遗传多样性

利用30个微卫星分子标记对海南鲤(HN)、长江鲤(CY)、月亮湖鲤(YL)、黑龙江鲤(FY)、呼伦湖鲤(ZL)、贝尔湖鲤(BR)6个野生鲤鱼群体的观测杂合度(Ho)、期望杂合度(He)、多态信息含量(PIC)和有效等位基因数(Ae)等进行了遗传检测,根据基因频率计算遗传相似系数和Nei氏标准遗传距离,χ2检验估计Hardy-Weinberg平衡,用近交系数(FST)和基因流(Nm)分析群体的遗传分化及其来源。同时,使用PHYLIP3.63软件绘制基于Nei氏标准遗传距离的UPGMA进化树。6个群体共检测到8,136个扩增片段,长度在125bp～414bp,30个基因座扩增出等位基因数从3～13个不等,共计210个等位基因,平均每个基因座扩增得到7个等位基因。结果显示:(1)6个野鲤群体的多态性指标均适中,多态信息含量依次0.44、0.52、0.53、0.57、0.63和0.64,有效等位基因数1.04～4.72个不等,平均有效等位基因数依次为2.19、2.60、2.42、2.43、2.45和2.33,无偏期望杂合度平均值为0.50、0.59、0.56、0.56、0.57和0.54;(2)遗传相似系数BR与ZL最高(0.8511),BR与HN最低(0.6688),聚类结果与地理分布呈一定相关性。     

Purification and Characterization of Serum Serpin from Carp (Cyprinus carpio)

A serine proteinase inhibitor, termed serpin62, was purified to homogeneity from carp serum with an increase in specific inhibitory activity of 6.2-fold and a 3% recovery rate after separation from α₁-antitrypsin. Specific inhibitory activity of serpin62 against bovine pancreatic trypsin was less than half of the specific antitryptic activity of α₁-antitrypsin. Under both reducing and nonreducing conditions, serpin62 was estimated to have a molecular weight (62,000) apparently larger than that of α₁-antitrypsin (55,000). They both consist of single polypeptide chains, but serpin62 differs from serine proteinase inhibitors from muscles of carp and white croaker in molecular weight and structure. Antibody raised against serpin62 immunologically crossreacted with serpin62 and had no crossreactivity with fish serum α₁-antitrypsin and muscular analogues. The antibody was susceptible to both serpin62 and its derivatives, which were widely distributed in carp tissues. Serpin62 is most likely distinct from other fish serine proteinase inhibitors expressing antitryptic activity physicochemically and immunologically. Received June 4, 1998; accepted September 10, 1998.     

Tissue-Specific Fatty Acids Response to Different Diets in Common Carp (Cyprinus carpio L.)

Fish depend on dietary fatty acids (FA) to support their physiological condition and health. Exploring the FA distribution in common carp (Cyprinus carpio), one of the world''s most consumed freshwater fish, is important to understand how and where FA of different sources are allocated. We investigated diet effects on the composition of polar and neutral lipid fatty acids (PLFA and NLFA, respectively) in eight different tissues (dorsal and ventral muscle, heart, kidney, intestine, eyes, liver and adipose tissue) of common carp. Two-year old carp were exposed to three diet sources (i.e., zooplankton, zooplankton plus supplementary feeds containing vegetable, VO, or fish oil, FO) with different FA composition. The PLFA and NLFA response was clearly tissue-specific after 210 days of feeding on different diets. PLFA were generally rich in omega-3 polyunsaturated FA and only marginally influenced by dietary FA, whereas the NLFA composition strongly reflected dietary FA profiles. However, the NLFA composition in carp tissues varied considerably at low NLFA mass ratios, suggesting that carp is able to regulate the NLFA composition and thus FA quality in its tissues when NLFA contents are low. Finally, this study shows that FO were 3X more retained than VO as NLFA particularly in muscle tissues, indicating that higher nutritional quality feeds are selectively allocated into tissues and thus available for human consumption.     

Carp (Cyprinus carpio) as a powerful invader in Australian waterways

1. The invasion of carp (Cyprinus carpio L.) in Australia illustrates how quickly an introduced fish species can spread and dominate fish communities. This species has become the most abundant large freshwater fish in south‐east Australia, now distributed over more than 1 million km². 2. Carp exhibit most of the traits predicted for a successful invasive fish species. In addition, degradation of aquatic environments in south‐east Australia has given them a relative advantage over native species. 3. Derivation of relative measures of 13 species‐specific attributes allowed a quantitative comparison between carp and abundant native fish species across five major Australian drainage divisions. In four of six geographical regions analysed, carp differed clearly from native species in their behaviour, resource use and population dynamics. 4. Climate matching was used to predict future range expansion of carp in Australia. All Australian surface waters appear to be climatically suitable for carp. 5. This assessment strongly reinforces the need for immediate management of carp in Australia to include targeted control of human‐assisted dispersal, such as use of carp as bait by anglers, distribution to new locations by anglers and the use of the ‘Koi’ strain in the aquarium industry. 6. Given their historical spread, dispersal mechanisms and ecological requirements, the expansion of carp across most of the remainder of Australia is to be expected.     

Cryoprotectants Reduce Protein Oxidation and Structure Deterioration Induced by Freeze-Thaw Cycles in Common Carp (Cyprinus carpio) Surimi

The objective of the present study was to investigate the effects of a cryoprotectant (a mixture of sucrose and sorbitol) on protein and lipid oxidation and structural changes in common carp (Cyprinus carpio) surimi subjected to different numbers of freeze-thaw (FT) cycles. Increased numbers of FT cycles were accompanied by increases in carbonyl content and thiobarbituric acid-reactive substances values and concomitant decreases in sulphydryl group (P<0.05) content, revealing that multiple FT cycles could cause protein and lipid oxidation in the carp tissue. Myofibrillar protein isolated from FT surimi also showed a reduced ATPase activity (P<0.05), increased hydrophobicity (P<0.05), and enhanced susceptibility to thermal aggregation. The addition of cryoprotectants significantly inhibited carbonyl formation and lipid oxidation (P<0.05), and its good protective effect on protein denaturation was demonstrated by its efficacy in maintaining Ca-ATPase activity, reducing the rapid exposure of hydrophobic and sulphydryl groups on the protein surface, and decreasing the degree of protein aggregation. Overall, the use of cryoprotectants reduced protein oxidation and structural deterioration induced by freeze-thaw in common carp surimi.     

烟酸诺氟沙星在松浦镜鲤体内的药动学特征

结合单纯聚集法和二步法,应用高效液相色谱(HPLC)技术研究了分别以10、30、60 mg/kg剂量对松浦镜鲤(Cyprinus carpio specularis)口灌烟酸诺氟沙星后,药物在实验鱼血浆中的药动学特征。3种给药剂量下,诺氟沙星在松浦镜鲤血浆中的血药浓度和时间关系均可用一级吸收二室开放模型进行描述,吸收半衰期(t1/2ka)分别为0.165、0.061、0.043 h,消除半衰期(t1/2β)分别为18.282、29.969、42.051 h,达峰时间(Tmax)分别为0.333、0.327、0.302 h,达峰浓度(Cmax)分别为4.780、6.247、12.689 mg/L,药时曲线下面积(AUC)分别为32.698、53.015、174.998 mg·h/L,表观分布容积(Vd)分别为1.044、4.347、4.561 L/kg。说明随着给药剂量的增加,诺氟沙星的吸收和消除速率均加快,给药剂量对药动学特征有显著影响。     

鲤鱼（Cyprinus carpio）生长激素基因的亚克隆及其序列分析

本文对ＰＣＲ扩增的６６８ｂｐ的ＤＮＡ片段进行了亚克隆,然后以Ｓａｎｇｅｒ双脱氧中止法为原理,利用美国ＡＢＩ公司３７０Ａ自动核酸序列分析仪,确定了６６８ｂｐ的核苷酸序列。序列分析表明鲤鱼生长激素基因的开放读框含有６３０ｂｐ,并推测其中包括２２个氨基酸的信号肽和１８８个氨基酸的成熟多肽。鲤鱼生长激素基因的酶切图谱和序列分析的结果都证明我们已获得了全长的鲤鱼生长激素基因。     

Evaluation of Acute Toxicity Levels and Ethological Responses Under Tetrachlorocatechol Exposure in Common Carp,Cyprinus carpio (Linnaeus)

Tetrachlorocatechol (TCC) is one of the most toxic chlorinated catechol produced by the chlorobleaching of pulp and frequently found in the kraft pulp mill effluents. Toxicity of TCC to common carp, Cyprinus carpio and their ethological changes were evaluated in the present study. The 24, 48, 72 and 96 h LC₅₀ values of TCC to C. carpio were 4.73, 3.95, 3.20 and 2.43 mg/l respectively. The mortality rate of C. carpio showed significant relationship (p < 0.05) at different exposure times (24, 48, 72 and 96 h) in different treatments of TCC (2, 3, 4, 5 and 7 mg/l). On the other hand, a significant correlation (p < 0.01) was observed between the mortality rate of fish with all the concentrations of TCC. Mean opercular movement in the fish increased significantly with the increasing dose and time of exposure to TCC indicating acute respiratory distress. The jerky movement, somersaulting, circular movement and mucus secretion of fish increased gradually with the increasing concentrations and exposure times of TCC for adapting a compensatory mechanism to derive energy in order to avoid stress due to toxicity.