盐度变化对黑鲷生长激素及其受体的影响

将海水鱼类黑鲷Sparusmacrocephalus从盐度为26.6的海水直接转入淡水,考察渗透压的急剧降低对黑鲷的生长激素(growthhormone,GH)及其受体(growthhormonereceptor,GHR)等生理指标的影响。研究发现:(1)黑鲷从海水直接转入淡水后48h有33%的鱼死亡,对照组无死亡现象;(2)黑鲷在转入淡水48h内其血清生长激素(GH)含量、肝脏及鳃中GHR及GHRmRNA含量与对照组均无显著差异;(3)黑鲷在转入淡水48h后其肾脏中GHR含量较对照组显著升高(p<0.05),肾脏中GHRmRNA含量在24h及48h显著高于对照组(p<0.05)。结果表明,从GH含量看,黑鲷从海水环境到淡水低渗环境与鲑鳟鱼类从淡水到海水环境的适应性调节机制有差异,而黑鲷肾脏中GHR的含量变化可能参与黑鲷对外界环境渗透压的这种急剧降低的适应性调节。     

Identification of aryl hydrocarbon receptor 2 in aquatic birds; cDNA cloning of AHR1 and AHR2 and characteristics of their amino acid sequences

The toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and its related planar halogenated aromatic hydrocarbons (PHAHs) are mediated by the aryl hydrocarbon receptor (AHR). To investigate the potential sensitivity to PHAHs and the evolutional diversity of AHR in aquatic birds, AHR cDNAs were initially cloned and sequenced from the livers of a black-footed albatross (Diomedea nigripes) and a common cormorant (Phalacrocorax carbo). In this study, we report the identification of two distinct AHR paralog genes in these species. The two full-length AHR cDNAs from albatross were highly divergent (33% overall amino acid identity, and 60% identity in the N-terminal half). Phylogenetic analysis showed that one of them belongs to the AHR1 clade and the other one to the AHR2 clade, which has been identified only from fishes, but not yet from mammals and birds. Albatross AHR1 encoded a 861-residue protein with a predicted molecular mass of 96.7 kDa, and in the case of albatross AHR2, 925 amino acids and 100.7 kDa. From cormorant liver, the full-length AHR1 cDNA and the partial AHR2 cDNA were cloned. This result strongly suggests that bird species also possess two distinct AHR genes (AHR1 and AHR2). To our knowledge, this is the first report on the presence of an AHR2-like isoform in bird species as well as in fish.     

棘皮动物感觉功能及其相关受体的研究进展

感觉系统是外界环境与有机体内在信号系统之间的“转换器”,也是生物应对外界环境变化的“哨兵”,能够敏锐捕捉外界环境信号的变化,在有机体的生长、摄食和繁殖等重要生物学过程中起到了至关重要的作用。棘皮动物具有特殊的五辐对称结构,缺乏神经中枢,主要依靠散布全身的感觉系统对外界进行感知并做出响应,其关键的分类地位和独特的生物学特征为感觉系统的功能研究提供了特殊的背景。本文对棘皮动物感觉系统研究方法、不同感觉系统介导的行为特征以及相关受体发掘进行了全面综述,以期进一步解析棘皮动物感觉系统在棘皮动物特殊生理行为调控中的作用机制,为棘皮动物关键经济物种的高效绿色增养殖提供科学的理论支撑。     

石斑鱼转录因子NR1D1和NR1D2在冷胁迫中的调控作用

NR1D1和NR1D2转录因子属于核受体转录因子家族,二者通过调控靶基因表达参与众多生物学过程,尤其在昼夜节律调节中发挥重要作用。本研究通过分析NR1D1和NR1D2的蛋白质分子特征以及在冷胁迫下珍珠龙胆石斑鱼(Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂)的基因调控关系以及它们所参与的通路,揭示它们在鱼类低温环境中的功能作用。生物信息学比较分析显示〖STBX〗NR1D1和NR1D2〖ST〗基因序列均有23个开放阅读框,编码的蛋白质均属于不稳定的亲水性蛋白质。NR1D1和NR1D2蛋白质均含有ZF C4类锌脂蛋白结构域和Hormone_Recep核激素受体的配体结合域,主要配体为含铁原卟啉IX。这两种蛋白主要在细胞核中发挥调控基因表达的作用,且与昼夜节律通路的多个关键基因有直接作用关系。结果表明,在冷胁迫下NR1D1和NR1D2作为转录因子可通过调控靶基因的表达,参与昼夜节律调节并促进糖脂代谢、线粒体氧化等过程,为珍珠龙胆石斑鱼提供必要的能量,有助于增强其耐寒性。     

The roles of thyroid hormone receptor and T3 in metamorphosis of Haliotis diversicolor

Thyroid hormone is a kind of important hormone which regulates metamorphosis. Its role is well described in amphibian metamorphosis. Thyroid hormones (T3 and T4) have also been demonstrated to play a role in metamorphosis of marine invertebrates. However, the mechanism of thyroid hormone in metamorphosis of marine invertebrates remains unknown. A homolog of vertebrate thyroid hormone receptor (TR) was cloned and identified in abalone Haliotis diversicolor and was named HdTR . The mRNA expressions of HdTR , thyroid peroxidase ( TPO ), thyroid peroxidase 1 ( TPO1 ), idothyronine deiodinase Ⅲ( IDⅢ) and integrin alpha-V ( ITGAV ) had significant diff erence in metamorphosis of H . diversicolor . Metamorphosis rate and mortality rate were significantly diff erent in HdTR RNAi experiment and T3 inducing experiment. In RNAi experiment, ITGAV and CCND1 (cyclin D1) expression of dsRNA HdTR exposing group were significantly lower than those of blank control and negative control. But CTNNB (catenin beta) expression of dsRNA HdTR exposing group was higher than that those of blank control and negative control. ERK (extracellular signal regulated kinases) and PI3K (phosphoinositide-3-kinase) had no significant diff erence in RNAi experiment. Moreover, ITGAV of 1 μmol/L T3 group was significantly lower than that of 0 μmol/L T3 group, PI3K expression of 10 μmol/L T3 group was higher than that of 0 μmol/L T3 group, and the other genes expression had no significant diff erence in T3 inducing experiment. The data of genes expression suggested that CCND1 might be an eff ector gene of TR genomic action, while CTNNB might be regulated by unliganded TR. CCND1 and CTNNB may be involved in cell proliferation of metamorphosis. T3 might regulate the expression level of PI3K via nongenomic way. These results shed light on the mechanism of thyroid hormone in abalone metamorphosis.     

缢蛏多巴胺受体基因及其在损伤修复中的作用

多巴胺受体(dopamine receptor)作为生物体中重要的神经递质受体, 在生物体的生长、发育、代谢等多个生理过程中都发挥着重要的功能。本研究从缢蛏Sinonovacula constricta转录组文库中筛选获得多巴胺受体基因的部分片段, 结合RACE(rapid-amplification of cDNA ends)技术和降落 PCR(polymerase chain reaction)技术, 克隆得到缢蛏两个多巴胺受体的选择性多聚腺苷酸化(alternative polyadenylation, APA)的变异体, 命名为ScDopR2a和 ScDopR2b, 长度分别为1824bp和2758bp。两个变异体均包含相同的5′非翻译区(untranslated regions, UTR)(24bp)和开放阅读框(open reading frame, ORF)(1440bp), 共编码479个氨基酸残基。但是两个变异体的3′UTR 长度不同, 分别为360bp和1294bp, 其中ScDopR2b在polyA尾前插入936bp。在同源的ORF区设计引物, 采用实时定量PCR分析多巴胺受体基因在不同组织中的表达特征, 结果表明多巴胺受体基因在水管、鳃、斧足的表达量显著高于其他组织。在该基因序列3′UTR插入片段区域设计特异引物检测ScDopR2b的组织表达情况, 结果表明在水管、鳃、斧足的表达量仍高于其他组织, 表达趋势与ORF区大致相同。进一步设计缢蛏组织损伤实验, 对进水管前端进行损伤处理, 在处理后的第4h、8h、12h、24h、48h、60h和72h取样, 荧光定量检测结果显示, 同源区表达量在12h和48h呈上调, 在8h、24h和72h下调, 且ScDopR2b表达趋势与同源区表达模式大致相同。研究结果表明, 缢蛏多巴胺受体参与了损伤修复过程, 其表达特征可能与多巴胺作为补偿性神经递质的作用有关。     

标记乳糖基白蛋白SPIO作为磁共振肝细胞受体显影剂的实验研究

目的是探讨标记乳糖基白蛋白的超顺磁性氧化铁粒子（ＬＡＣ－ＨＳＡ－ＳＰＩＯ）作为配体与肝细胞膜去唾液酸糖蛋白受体（ＡＳＧ）特异性结合可能性;评价ＬＡＣ－ＨＳＡ－ＳＰＩＯ增强磁共振受体成像对大鼠微小肝癌灶的显示能力。方法：将Ｆｅ３＋Ｆｅ２＋混合液与ＬＡＣ－ＨＳＡ在碱性条件下制备出ＬＡＯ－ＨＳＡ－ＳＰＩＯ;应用　二乙基亚硝酸胺建立大鼠肝癌模型;电镜及Ｐｅｒｌｓ（普鲁士蓝）染色,观察ＬＡＣ－ＨＳＡ－ＳＰＩＯ在肝组织的颁;并比较ＬＡＣ－ＨＳＡ－ＳＰＩＯ增强前后大鼠肝癌的显示情况。得到结果是（１）Ｐｅｒｌｓ染色证实有ＬＡＣ－ＨＳＡ－ＳＰＩＯ分布到肝细胞浆内;电镜下也显示肝细胞膜表面及胞浆内有电子致密ＳＰＩＯ粒子分布;（２）注射小剂量ＬＡＣ－ＨＳＡ－ＳＰＩＯ可观察到明显明显肝脏强化效果,大鼠肝癌检出率由平扫３６．８％（７／１９）提高到７８％（１５／１９,Ｐ＜００１）。结论：ＬＡＣ－ＨＳＡ－ＳＰＩＯ是一种新型的具有肝细胞ＡＳＧ受体靶向性显影剂,对提高肝脏微小瘤灶检出率有章要价值。     

三疣梭子蟹(Portunus trituberculatus)RXR基因克隆及其在蜕皮过程中的表达分析

本研究通过转录组测序和RACE技术克隆了三疣梭子蟹类维甲酸X受体2(PtRXR2)基因cDNA全长(登录号:KF914662),并通过荧光定量PCR(qRT-PCR)技术研究了该基因在三疣梭子蟹不同组织及不同蜕皮阶段的表达情况。结果表明,(1)PtRXR2基因cDNA全长1718bp,包括5’非编码区(5’-UTR)141bp、3’-UTR 209bp和开放阅读框1368bp,编码455个氨基酸,预测分子量和等电点为49.75kDa和6.79。(2)PtRXR2推导氨基酸序列的Blastp结果显示,PtRXR2与已知甲壳动物RXR的一致性为74%—99%,系统进化树分析表明PtRXR2与其它甲壳动物RXR聚为一支。(3)qRT-PCR结果显示PtRXR2在C期三疣梭子蟹Y器官、鳃、眼柄和大颚器中表达水平较高,肝胰腺中的表达水平最低。其它6个组织中的表达水平居中。(4)不同蜕皮阶段,PtRXR2在Y器官、眼柄和胸神经中均是AB期表达水平最高,E期最低,这可能与上述器官的细胞增殖主要发生在AB期有关;肌肉中的PtRXR2表达水平在AB和C期较高,E和D期最低,与肌肉的生长和营养物质积累主要发生在这两个阶段相对应;肝胰腺中的PtRXR2表达水平在AB期最高,C期最低,这暗示PtRXR2主要参与肝胰腺中的上皮细胞增殖和分化,对于C期的营养代谢调控可能不起关键作用。     

Towards molecular understanding of species differences in dioxin sensitivity: initial characterization of Ah receptor cDNAs in birds and an amphibian

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and related planar halogenated aromatic hydrocarbons (PHAHs) are highly toxic to most vertebrate animals, but there are dramatic species differences in sensitivity, both within and among vertebrate classes. For example, studies in cultured avian hepatocytes have revealed differential sensitivity of birds to PHAHs [Kennedy et al. (1996). Toxicol. Appl. Pharmacol., 141, 214-230]. Differences in the characteristics or expression of the aryl hydrocarbon receptor (AHR) could contribute to these species differences in PHAH responsiveness. To investigate the molecular mechanism of differential PHAH sensitivity, we have begun to characterize the AHR in white leghorn chicken (Gallus gallus), Pekin duck (Anas platyrhynchos), and common tern (Sterna hirundo), as well as an amphibian, mudpuppy (Necturus maculosus). Partial AHR cDNAs encompassing the helix-loop-helix and PAS domains were cloned and sequenced. Comparison of amino acid sequences in this region indicated a high degree of sequence conservation among the bird species (97% amino acid identity). The percent identity between bird sequences and either mouse or mudpuppy was lower (79%); the mudpuppy AHR was 74% identical to the mouse AHR. Phylogenetic analysis of these and other AHR amino acid sequences showed that the bird and mudpuppy AHRs were more closely related to mammalian and fish AHR1 forms than to fish AHR2. Future studies include the in vitro expression and functional characterization of AHRs from these and other non-mammalian vertebrates.     

有机磷农药对胆碱能系统影响研究进展

有机磷农药具有高效、低毒等特点被广泛应用于农业生产,其残留对生物和人类的健康构成了潜在的威胁.由于有机磷农药主要通过影响胆碱能系统的活性发挥其神经毒性作用,因此本文综述了有机磷农药对胆碱能系统中乙酰胆碱脂酶、胆碱乙酰转移酶、毒蕈碱型受体和烟碱型受体产生的影响及可能的致毒机理.有机磷农药首先抑制乙酰胆碱脂酶的活性产生神经...