性类固醇激素E2、MT 对尼罗罗非鱼(Oreochromis niloticus)雌、雄生长差异的影响

性类固醇激素对动物生长有一定的影响, 但其对鱼类雌、雄生长差异的影响机理尚不清楚。 为进一步了解动物雌雄生长差异的内在原因, 本文通过在体注射和荧光定量PCR 方法, 比较了性类 固醇激素(E₂、MT)对尼罗罗非鱼雌鱼和雄鱼的生长及对垂体GH、肝脏GHR1、IGF-I mRNA 表达的 影响。结果表明: 性类固醇激素E₂ 和MT 对尼罗罗非鱼的生长及生长轴相关基因的表达均有显著的 影响, 并且这种影响存在明显的性别二态性。E₂ 明显促进雌鱼的生长但不显著影响雄鱼的生长, MT 显著促进雄鱼和雌鱼的生长; E₂ 可使雌鱼垂体GH 和肝脏GHR1 、IGF-I 的mRNA 水平显著升高 (P<0.05), 但降低雄鱼垂体GH和肝脏GHR1的mRNA表达; MT 显著提高雄鱼垂体GH和肝脏GHR1、 IGF-I mRNA 水平(P<0.05), 但降低雌鱼垂体GH 的mRNA 表达, 促进雌鱼肝脏GHR1 mRNA 的表达, 但对雄鱼肝脏GHR1 的促进作用明显大于雌鱼(P<0.05).上述结果表明, 性类固醇激素对不同性别尼 罗罗非鱼的生长及生长轴相关基因的mRNA 表达有不同的影响, 表现出明显的性别二态性, 提示不 同性别鱼体内性类固醇激素水平的不同可能是导致尼罗罗非鱼雌雄生长二态性的另一内在原因。     

活性菌肽豆蛋白对鲫(Carassius auratus) 促生长的作用效果及机理分析

采用不同饲料配方投喂,研究了普通豆粕(对照组)、发酵豆粕和活性菌肽豆蛋白配合饲料对鲫(Carassius auratus)生长性状影响的研究。结果表明,活性菌肽豆蛋白组的相对体重生长率高于对照组41.87%(P<0.01)和发酵豆粕组19.11%;血清中的三碘甲状腺原氨酸(T3)、甲状腺素(T4)、胰岛素样生长因子-I(IGF-I)和生长激素(GH)含量活性菌肽豆蛋白组最高,其次是发酵豆粕组,而对照组最低;实时定量PCR检测显示:IGFs(IGF-I和IGF-II)基因的表达量在活性菌肽豆蛋白组中最高,其次是发酵豆粕组,对照组的表达量最低。GHR基因的表达量在活性菌肽豆蛋白组和发酵豆粕组间无显著性差异,但明显高于对照组。因此,活性菌肽豆蛋白可能通过调控鲫血清GH、IGF-I、T3和T4等激素水平,及肝脏GHR和IGFs基因的表达,进而影响鲫的生长。     

半胱胺盐酸盐(CSH)对尼罗罗非鱼(Oreochromis niloticus)生长及生长轴相关基因表达的影响

为了分析半胱胺盐酸盐(CSH)对尼罗罗非鱼生长调节的作用,设计了长期和短期2个实验,采用腹腔注射(剂量100μg/g体重)方法,分析CSH对尼罗罗非鱼绝对生长率、特定生长率、肝体系数和肥满度的影响,并应用荧光实时定量PCR方法检测在注射CSH后不同时段(6h,12h,24h,2周)尼罗罗非鱼垂体GH、肝脏GHR和肝脏IGF-I基因的表达变化。结果表明,CSH组尼罗罗非鱼的绝对生长率、特定生长率、肝体系数、肥满度均显著高于对照组(P0.05);注射CSH后12h、24h垂体GH和肝脏GHRmRNA表达水平均显著升高(P0.05),2周后恢复到对照组水平;注射CSH后6h肝脏IGF-ImRNA表达水平显著升高(P0.05),12h恢复到对照组水平,24h和2周表达水平极显著上升(P0.01)。以上结果提示,CSH可显著上调尼罗罗非鱼生长轴相关基因的表达,从而促进鱼类的生长。     

硬骨鱼类神经内分泌-免疫网络及其调控研究

综述了近10余年来国内外有关硬骨鱼类神经内分泌-免疫网络(N-E-IN)及其调控的研究概况。以下丘脑-垂体-肾间组织(HPI)轴为核心,阐述了下丘脑肽类激素、垂体激素、糖皮质醇(GC)、性类固醇激素(sexual steroid)对鱼类免疫功能的调节,环境胁迫(stress)对鱼类(N-E-IN)的影响,细胞因子(cytokine)对硬骨鱼类内分泌功能的影响,展望了今后该领域的研究方向。     

鱼类生长激素的分子生物学和应用研究的进展

依据１９８５－１９９５年国际有关资料对鱼类生长激素的分子生物学及其研究的最新成果与进展进行综合评述。研究表明，已完成鳗鲡等９种鱼类生长激素的氨基酸组成与全序列分析及大麻哈鱼等２０种鱼类的生长激素基因的分离与克隆，有些已在工程菌或动物细胞中的高效表达；鱼类生长激素基因的调控机制与生长激素的活性部位研究正在深入进行；将外源的生长上入鱼体或将外源的生长激素基因导入受体鱼，以期促进生长，增加抗性，是生长激     

盐度调控对花鲈(Lateolabrax maculatus)生长相关基因表达的影响

生长激素受体(GHR)作为GH/IGF轴的中心环节,在内分泌调控中发挥重要作用。本实验采用c DNA末端快速扩增法(RACE)技术克隆出花鲈GHR1和GHR2的c DNA全长序列。急性低盐度调控实验设为海水组、半海水组和淡水组。测定了急性低盐度调控24、48、96、144、192h后,花鲈肝脏中GHRs、IGF-1及垂体GH表达情况。结果表明,GHR1 c DNA全长序列2436bp,编码637个氨基酸;GHR2 c DNA全长序列2940bp,编码582个氨基酸。GHR1与GHR2由信号肽、胞外区、跨膜区、胞内区组成,且结构存在差异。脑、肾、鳃中GHR1表达明显高于GHR2;而在肌肉、垂体、肝脏、盲肠、胸腺、心脏中,GHR2表达明显高于GHR1。24h时,各组GHR1表达不变,GHR2、GH、IGF-1显著下降。之后,相对于海水组,淡水组和半海水组GHRs和IGF-1表达升高,而GH下降,GH与GHR负相关。据结果推测,花鲈GHR2可能为SL受体,GH/IGF轴参与低渗调控可能是通过增加GHRs,进而激活下游IGF-1表达而实现。     

植物雌激素对鱼类影响的研究进展

植物雌激素是一类植物来源的具有雌激素活性的非甾族杂环多酚类化合物,也是一类潜在的环境内分泌干扰物,可以影响鱼类的生理、生殖等正常活动.本文介绍了植物雌激素的分布与来源,综述了各种植物雌激素对鱼类生长、行为和生殖的影响,并从植物雌激素与受体结合、抑制类固醇类激素的代谢、酶调节等几种可能的途径,讨论了植物雌激素对鱼类内分泌扰乱作用的机理.     

牙鲆2种IGFBP基因的组织表达和激素调控分析

在脊椎动物中,胰岛素样生长因子结合蛋白(IGFBPs)对于调节IGFs生物活性、调控机体的生长、代谢和繁殖有着非常重要的作用。但目前关于鱼类IGFBPs的研究还不是非常深入。本实验利用RT-PCR技术分析了牙鲆IGFBP-3和IGFBP-4基因在不同成体组织中的表达模式。结果显示,这2种基因在成体组织中均表达,其中IGFBP-3基因在性腺中表达量最高,在肝脏中最低;而IGFBP-4基因在肠中含量最丰富。利用原代培养的牙鲆肝细胞研究了不同激素对牙鲆IGFBPs的调控作用。结果显示:生长激素、胰岛素和IGF-I能够以剂量依赖和时间依赖的方式调控IGFBP-3和IGFBP-4基因的表达。本研究结果表明,牙鲆IGFBP-3和IGFBP-4基因可以在体内不同组织中发挥作用,且不同激素的刺激可以调节它们的体外表达模式。     

银鲳(Pampus argenteus)胰岛素样生长因子-I基因的克隆和表达分析

胰岛素样生长因子-I(Insulin-like growth factor-I,IGF-I)是影响脊椎动物生长、发育及代谢的重要调控因子。本研究采用RT-PCR和RACE技术,克隆了银鲳(Pampus argenteus)肝脏IGF-IcDNA序列,应用半定量RT-PCR、Real-time q PCR和原位杂交的方法检测了IGF-I的组织表达特性、在肝脏中的生长表达特性和IGF-I基因在肝脏中的定位。序列分析表明,IGF-I cDNA序列全长836bp,其5′非编码区128bp、3′非编码区92bp,开放阅读框(open reading frame,ORF)605bp,由此推导IGF-I前体蛋白由201个氨基酸组成;前体肽由信号肽、成熟肽、E肽三部分组成,其中信号肽59个氨基酸,成熟肽68个氨基酸,E肽74个氨基酸;成熟肽由B、C、A、D四个区域组成,E肽分析表明,银鲳IGF-I属Ea-4型。同源性比较结果表明,银鲳与同目鲈形目鱼类的IGF-I编码序列同源性较高,为83.52%—91.40%;与哺乳类、鸟类和爬行类的同源性较低。半定量RT-PCR和Real-time q PCR组织特异性表达结果显示,IGF-I m RNA在肝脏组织中的表达量最高,显著高于其它组织,肾脏、心脏、肌肉、脑、鳃、小肠、卵巢次之,嗅球、脾脏和胃中表达较低;半定量RT-PCR和Real-time q PCR不同生长阶段表达结果显示,IGF-I m RNA在30—50g肝脏组织中表达量最高(P0.05);IGF-I m RNA在肝脏中的原位杂交定位结果显示,在肝脏细胞中均有表达,阳性信号主要位于细胞质中,靠近细胞边缘处信号较强。     

环境内分泌干扰物诱导鱼类产生卵黄原蛋白的研究进展

目前已经被确认的环境内分泌干扰物达100余种,其中能够诱导鱼类产生卵黄原蛋白(Vitellogenin, VTG)的环境内分泌干扰物包括农药、多氯联苯类化合物、邻苯二甲酸酯类化合物等几大类.由于环境内分泌干扰物结构的复杂性,其诱导机制成为近年研究的热点.本文综述了能够诱导鱼类产生VTG的环境内分泌干扰物的种类、筛选方法及作用机制.     

鱼类生长激素的分离、鉴定及其生理功能研究的进展

主要对８０年代末至９０年代中期国际有关鱼类生长激素的分离，鉴定和生理功能研究的进展予以综述，资料表明，蓝鲨，罗非鱼等２１种鱼类的生长激素已被分离，并阐明鱼类ＧＨ一般是在碱性条件下，将脑垂体匀浆，或取其体外培养的分泌液，经凝胶过滤和反相高效液相色谱纯化得到。     

外源污染物对硬骨鱼甲状腺干扰作用机制的研究进展

甲状腺激素在鱼类生长发育、变态、生殖、渗透调节以及摄食和营养代谢中发挥着重要作用,外源化合物对硬骨鱼的甲状腺干扰作用会导致发育和孵化延迟、运动性降低。本文首先对鱼类甲状腺激素的中枢调控、合成、转运、代谢和发挥作用的相关过程进行简要介绍,随后从核受体介导途径、非受体介导途径两个方面综述了外源污染物对硬骨鱼甲状腺的干扰作用机制,最后从物种特异性、检测指标、膜受体介导途径三个方面提出了目前研究的不足之处以及解决方法,以期为今后探究外源污染物对鱼类甲状腺系统干扰作用机制的研究提供更多的参考。     

Reproductive life history stages sensitive to oil and naphthalene in Atlantic croaker

The reproductive effects of chronic exposure to water-soluble fractions (WSF) of diesel fuel oil and naphthalene in female Atlantic croaker during critical periods of their reproductive life history were investigated. Fish were repeatedly exposed to WSF of oil (2.5 or 5%) or naphthalene (0.5 or 1.0 ppm) for 5 or 8 weeks. Both pollutants blocked sexual maturation in some fish and impaired ovarian recrudescence in others. The majority of the oocytes in exposed fish were undeveloped and widespread oocyte atresia was evident at higher concentrations of these polycyclic aromatic hydrocarbons (PAH). The disruption of ovarian growth and oocyte development in croaker after PAH exposure was associated with alterations in reproductive endocrine function at multiple sites on the hypothalamus-pituitarygonadal axis. A third period of increased sensitivity to xenobiotics was observed during final oocyte maturation.     

Growth hormone effect on xenobiotic-metabolizing activities of rainbow trout

In recent years several studies reported the regulation by growth hormone (GH) of the expression of a variety of P450 forms in mammals. However the effect of GH on the xenobiotic-metabolizing enzymes of fish are still unknown. The aim of this work was to investigate the effects of ovine GH—a growth hormone known to be efficient in trout—on the cytochrome P450 level and on aryl hydrocarbon hydroxylase, aminopyrine-N-demethylase, glucuronyl transferase and glutathione transferase activities in trout. The GH-implanted trout (n = 50) each received a single cholesterol pellet containing ovine GH and were compared to control animals (n = 50) receiving a single cholesterol pellet without GH. After 15 days fish were killed and the liver and gills were excised for the measurement of xenobiotic-metabolizing enzyme activities. GH treatment significantly decreased the level of hepatic cytochrome P450 and the activities of cytochrome P450 dependent monooxygenases. In contrast, no significant effect of the treatment was observed on the glutathione transferase and UDP-glucuronyl transferase activities. Moreover, GH treatment had no effect on branchial phase I and phase II enzyme activities. This study provides evidence that GH level significantly affects the expression of several members of the hepatic cytochrome P450 family in trout.     

Reproductive endocrine function in female atlantic croaker exposed to pollutants

To examine whether xenobiotics impair teleost reproduction by altering reproductive endocrine function, steroid hormone secretion and ovarian growth were investigated in female Atlantic croaker (Micropogonias undulatus) after chronic oral administration with sublethal doses of three classes of reproductive toxins (lead, benzo[a]pyrene and Aroclor 1254). All the sublethal treatments significantly impaired ovarian growth as assessed by the gonadosomatic index. Reduced ovarian growth was accompanied by a significant decline in circulating 17β-estradiol levels in fish exposed to lead and benzo[a]pyrene. All three toxicants significantly decreased plasma testosterone levels. However, the steroidogenic capacity of ovarian tissue in vitro was not altered by xenobiotic exposure. The data suggest that the decreased ovarian growth in croaker after pollutant exposure may be a consequence of the decline in plasma 17β-estradiol levels. Further, this decrease in circulating levels of gonadal steroids does not appear to be caused by a direct effect of the chemicals on ovarian steroid synthesis.     

Intersexes in Mississippi River shovelnose sturgeon sampled below Saint Louis, Missouri, USA.

The Missouri Department of Conservation detected high organochlorine levels in tissues of fish from the Mississippi River south of Saint Louis. Chlordane, which is both a carcinogen and an endocrine disrupter, reached 2960 micrograms/kg in fillets of shovelnose sturgeon, Scaphirhynchus platyorynchus, and 1926 micrograms/kg in their roe, wet weight basis. Polychlorinated biphenyls (PCBs) and p,p'-dichlorodiphenyldichloroethylene (DDE) in roe have reached 5810 and 780 micrograms/kg, respectively. The purpose of this study was to determine if endocrine disrupters and carcinogens are a potential cause of the significantly diminished reproduction capacity and population declines in the endangered pallid sturgeon, Scaphirhynchus albus. Twenty-four surrogate shovelnose sturgeons were sampled north of Saint Louis where contaminant levels were lower compared with 17 sampled south of Saint Louis where contaminant levels were high. Liver and gonad tissues were formalin-fixed, histologically processed and examined. Neoplasms were not found. The principal finding was that the testes of two of the seven downstream male fish contained ovigerous lamellae, in addition to mature sperm. Since sturgeons are gonochoristic and intersexes are rare and unusual, the occurrence of intersex in 29% of the male Mississippi River shovelnose sturgeon examined from a population contaminated with chlordane and other chemicals raise concern for human health, species biodiversity and preservation of waterway quality. These findings, however, need to be corroborated by expanded studies with a higher sample size.