南移养殖刺参(Apostichopus japonicus)原肌球蛋白基因的原核表达研究

本研究克隆和表达了刺参原肌球蛋白(Tropomyosin,TRP)基因,进一步研究刺参再生过程中重要分子的功能.结果表明,TRP基因序列总长为1203bp,5 ′非翻译区为105bp,3 ′非翻译区为240bp,该序列包含一个855bp的开放阅读框(open reading frame,ORF),编码284个氨基酸,分子量为33.27kDa,等电点为4.56.利用Escherichia coli对TRP进行了体外重组表达,在1mmol/L IPTG和37℃条件下诱导,能产生分子质量约为38kDa的重组蛋白,Western blot证明重组TRP与鼠抗刺参原肌球蛋白的多克隆抗体能特异性结合.     

用免疫细胞化学方法研究文昌鱼卵子——成熟前后原肌球蛋白的分布及变化

本文用免疫细胞化学方法对文昌鱼原肌球蛋白在成熟前后的卵子中的分布及变化进行了观察和研究,并将其在亚微结构上进行了定位。结果表明:在卵巢中的较大的未成熟的卵母细胞上就已有原肌球蛋白的存在,主要分布在内滤泡层及卵内的某些卵黄颗粒上,皮质颗粒上也存有少量;在成熟卵中,原肌球蛋白则大量存在于所有的卵黄颗粒上,皮质颗粒也有少量存在;在量上是由动物极到植物极逐渐递增,这和卵黄颗粒的分布是一致的。     

脊尾白虾肌球蛋白重链和肌球蛋白轻链基因的克隆与表达分析

肌球蛋白重链和肌球蛋白轻链是粗肌丝的重要组成单位,其表达量的高低影响肌纤维的组成和肌肉生长。为了解析肌球蛋白重链(Myosin heavy chain,MHC)和肌球蛋白轻链(Myosin light chain,MLC)基因在脊尾白虾(Exopalaemon carinicauda)生长发育中的作用,采用RACE技术,克隆了脊尾白虾EcMHC2、EcMLC1全长cDNA序列,采用实时荧光定量PCR方法分析了EcMHC2、EcMLC1在脊尾白虾不同组织、不同幼体发育时期及大小差异个体在不同生长时期的表达特征。研究显示,脊尾白虾EcMHC2基因(GenBank:MG545144)全长5 929bp,开放阅读框5 727bp,编码1 909个氨基酸;脊尾白虾EcMLC1基因(GenBank:MG545145)全长1 506bp,开放阅读框459bp,编码153个氨基酸。脊尾白虾肌球蛋白重链和肌球蛋白轻链在系统进化上与其它十足目甲壳动物的关系较近。脊尾白虾EcMHC2、EcMLC1在各组织中均有表达,但主要在肌肉组织内表达。在脊尾白虾幼体发育的不同时期,EcMHC2、EcMLC1在溞状幼体Ⅱ期的表达量显著高于溞状幼体的其它时期;仔虾第一天,EcMHC2、EcMLC1的表达量显著高于其它时期。在脊尾白虾生长的四个阶段中,大个体组EcMHC2的表达量均显著的高于小个体组;60、80和120日龄时期,大个体组的EcMLC1表达量显著的高于小个体组;在大个体和小个体组中,EcMHC2、EcMLC1在80日龄的表达量最高。脊尾白虾EcMHC2、EcMLC1在大个体组与小个体组的差异表达提示它们在脊尾白虾生长发育过程中发挥重要作用。     

仿刺参肠道神经定位初步研究

本文以仿刺参(Apostichopus japonicus)肠道组织为研究材料,利用HE染色、米利根染色和免疫组化技术对其组织学结构及神经定位进行了观察和分析,初步探讨了仿刺参肠道的神经分布。HE染色结果表明,仿刺参肠壁由外向内分别为:体腔上皮层、肌肉层、内部结缔组织层和肠腔上皮层;和后肠相比,其中肠内壁常形成高而窄,且方向单一的褶皱,在结构上扩大了吸收面积,与其主要起消化吸收作用这一功能一致。米利根染色表明肠壁的纵向肌肉、肠腔上皮层和体腔上皮层均有神经成分存在,棘皮动物神经特异性抗体1E11的定位研究结果同样显示其免疫活性广泛而强烈,阳性位点集中于体腔上皮层、肌肉层和内部结缔组织层,且在肌肉层中表达最强,推测其可能参与肠道的再生过程及长期夏眠萎缩后的肠道快速修复过程。本研究为仿刺参神经生物学的研究提供了理论支持,同时也为仿刺参特殊生理机制的研究提供神经学参考资料。     

盐度驯化对虹鳟鳃、肌肉和肝脏非极性脂脂肪酸的影响

本文分析了盐度驯化前后虹鳟(Oncorhynchus mykiss)鳃、肌肉和肝脏中的非极性脂脂肪酸组成。研究显示,盐度驯化前后虹鳟3种组织器官的非极性脂脂肪酸种类组成没有变化,但非极性脂脂肪酸比例组成出现了组织特异性变化。盐度驯化对虹鳟鳃的非极性脂脂肪酸组成影响不显著,但肌肉和肝脏的非极性脂中饱和脂肪酸(SFA)比例显著下降;肌肉和肝脏C18多不饱和脂肪酸(C18PUFA)、亚麻酸族不饱和脂肪酸(n-3PUFA)和亚油酸族不饱和脂肪酸(n-6PUFA)比例显著上升,非极性脂脂肪酸的不饱和指数(UI)和不饱和率(U/S)显著上升。本研究表明,在盐度驯化中,鳃不是虹鳟主要的能量供给器官,肌肉和肝脏可以分解非极性脂的饱和脂肪酸为虹鳟适应盐度环境供能,是主要的能源供给器官;虹鳟肝脏中C18PUFA的升高可加快内源性LC-PUFA的合成,有利于虹鳟对海水环境的适应和自身的生长发育。     

饥饿和复投喂对大黄鱼(Larimichthys crocea)IGF-Ⅰ、mTOR、MyoD和MHC基因表达的影响

为从分子水平研究饥饿对大黄鱼肌肉生长代谢的影响,本实验采用qPCR技术研究了在饥饿胁迫以及复投喂过程中胰岛素样生长因子基因IGF-Ⅰ（insulin-like growth fator-Ⅰ）、雷帕霉素靶蛋白基因mTOR（mammalian target of rapamycin）、成肌分化抗原基因MyoD（myogenic differentiation antigen）和肌球蛋白重链基因MHC（myosin heavy chain）等4个大黄鱼肌肉生长调控相关基因在肝脏、脾脏、脑、心脏、肠、鳃、肌肉和肾8个组织中的表达模式。结果显示：四个基因在正常大黄鱼不同组织间表达存在显著差异;饥饿显著降低IGF-Ⅰ在肝组织中的表达量（P<0.05）,在复投喂14d时显著上升,此外在肠和鳃组织中表达量变化显著（P<0.05）;mTOR表达量随着饥饿时间的延长,在脾、心和肾组织中表达量下降,在脑、鳃和肌肉组织中表达量显著升高（P<0.05）;MyoD在饥饿胁迫和恢复投喂期间,在肝脏、鳃和肌肉组织中表达量变化极显著（P<0.01）;饥饿和恢复投喂对MHC在鳃和肌肉中的表达影响显著（P<0.05）。结果提示饥饿可能通过调节这些肌肉生长相关基因的表达来影响肌肉的生长。     

刀额新对虾主要过敏原蛋白的肽指纹图谱鉴定与分析

以我国常见的刀额新对虾为研究对象,旨在获得其主要过敏原的氨基酸序列等基础数据,增加对虾类过敏原的认识和理解。分离和纯化了分子量为36 kD的虾主要过敏原蛋白,利用激光辅助解析/飞行时间质谱对其进行肽质量指纹图谱鉴定,并对鉴定的结果采用Mascot搜索引擎在NCBInr数据库上进行搜索。结果表明刀额新对虾主要过敏原蛋白与斑节对虾原肌球蛋白匹配分值最高为268,吻合肽段27条,序列覆盖率为65%;与其它无脊椎动物如腐食酪螨、衣鱼等的原肌球蛋白的序列覆盖率也很高,分别达到了51%和53%。这一结果不仅表明了刀额新对虾与其它甲壳类海产品过敏原存在着高度同源性,而且为其与甲壳类及其它无脊椎动物主要过敏原之间存在严重交叉反应的现象提供了理论依据。     

《海洋与湖沼》2010 年第6 期论文导读

<正>南移养殖的刺参(Apostichopus japonicus)cDNA文库的构建及原肌球蛋白基因的研究以南移养殖的刺参(Apostichopus japonicus)为实验材料,采用非均一化的Oliga-dT引物定向克隆技术构建了南移刺参混合组织的cDNA文库。结果表明,cDNA文库的     

花鲈Argonaute基因家族的快速进化与表达分析

RNA干扰(RNA interfering,RNAi)是广泛存在于动植物等生物中的一种高度保守、序列特异的RNA降解机制.Argonaute蛋白是RNAi途径的关键组成部分,其结构、功能和进化模式在无脊椎动物中已得到广泛研究,但在脊椎动物特别是硬骨鱼中的报道还十分有限.本研究从已测序物种的基因组中鉴定了12种哺乳动物和...     

心跳的秘密

是什么控制了心跳？实验证明，心跳的过程并没有我们想象得那么复杂。心脏细胞中钙离子的突然增加使细胞中的大量纤维互相“拉扯”、收缩，进而引发心跳。这些纤维由两种蛋白组成肌动蛋白和肌球蛋白。在纤维收缩发生之前，肌动蛋白必须首先被激活。过去，人们认为钙离子和肌球蛋白共同参与了肌动蛋白的激活过程。伦敦大学国王学院的研究者发现，激活肌动蛋白似乎只有钙离子而已。     

Detection of DNA damage in mussels and sea urchins exposed to crude oil using comet assay

The single-cell microgel electrophoresis assay or the comet assay was used to evaluate DNA damage of dispersed crude oil on sea urchins (Strongylocentrotus droebachiensis) and mussels (Mytilus edulis L.). Sea urchins were exposed to 0.06 and 0.25 mg/L dispersed crude oil in a continuous flow system, while the mussels were exposed to 0.015, 0.06 and 0.25 mg/L dispersed crude oil. Sea urchin coelomocytes and mussel haemocytes were sampled after 4 and 5 weeks exposure, respectively. In the sea urchin coelomocytes, there was a significant concentration-related increase in the percentage of DNA in comet tail. In mussel haemocytes, there was a significantly higher percentage of DNA in comet tail for all treatments compared to the control. The responses were concentration-related up to 0.06 mg/L oil. The two highest exposure concentrations of mussels were not significantly different from each other. These results indicate that the comet assay can be used for biomonitoring of DNA damage in marine invertebrates following oil contamination.     

中间球海胆人工育苗附着基改良及不同规格种苗养殖效果的比较

人工育苗是我国培育中间球海胆养殖业种苗的主要手段,然而传统的海胆附着基的人工育苗效果不尽如人意,尤其是在控制稚胆剥离损伤方面。目前,海上吊笼养殖是中间球海胆人工养殖最主要的养成方式之一,而随着室内工厂化养殖技术的更新和优化,海胆的室内工厂化养殖产业发展迅速并受到广泛关注。为探明以软质塑料膜作为中间球海胆附着基的育苗效果以及不同规格种苗养殖的效果,分别使用硬质波纹板和软质塑料膜作附着基进行中间球海胆人工育苗,并开展了大、小两种规格（壳径分别为0.5 cm和2.0 cm）的海胆种苗室内工厂化养殖和海区养殖研究,旨在明确软质塑料膜替代传统硬质波纹板作海胆育苗附着基的可行性以及中间球海胆室内工厂化养殖和海区养殖效果的优劣。实验结果表明：软质塑料膜在采苗效率方面与波纹板无异,但在促进稚胆生长和提高存活率方面优势显著（p<0.05）;在不同规格中间球海胆种苗的工厂化养殖和海上养成研究中,相同养殖条件下,养殖12月龄的大规格种苗相较于小规格种苗在生长和存活方面优势显著（p<0.05）,同时工厂化养殖能确保更高的养殖成活率和更优的生长性能（p<0.05）,然而中间球海胆的工厂化养殖在...     

紫海胆生殖腺和棘壳中的金属元素

在生殖季节从大亚湾采集紫海胆Anthocidariscrassispina ,分离出生殖腺和棘壳 ,用ICP AES法测定其金属元素含量。结果表明 ,紫海胆生殖腺中含有Ca ,Mg ,Sr,Fe ,Mn等1 8种金属元素。这些元素几乎都同样在棘壳中存在 ,而且棘壳中的浓度比生殖腺的高出几十倍到几百倍。同时发现棘壳中金属元素的浓度比海水中的高很多 ,如Fe的浓度为海水的34万倍 ,Mn为 1 4.5万倍 ,说明紫海胆对海水中金属元素有很强的富集作用。结果还表明紫海胆生殖腺中金属元素的含量存在性别差异 ,雌性生殖腺中Zn含量是雄性生殖腺的 4倍 ,而雄性生殖腺中Mg的含量比雌性生殖腺高。以食品卫生标准来衡量 ,大亚湾紫海胆生殖腺金属元素含量完全符合国家标准GBn 2 38 84。如何开发和利用这一海产资源 ,值得深入研究。     

Low densities of sea urchins influence the structure of algal assemblages in the western Mediterranean

Numerous studies of interactions between urchins and algae in temperate areas have shown an important structuring effect of sea urchin populations. These studies focused almost wholly on the effect of high urchin densities on laminarian forests. In contrast, algal communities below 5–6 m depth in the northwestern Mediterranean are characterised by low sea urchin densities (<5 ind m⁻²) and the absence of laminarian forests. No previous research has addressed sea urchin/algal interactions in this type of community. To determine the effect of the most abundant echinoid species, Paracentrotus lividus, on well-established algal communities in this area, we performed a removal–reintroduction experiment in rocky patches located between 13 and 16 m depth in the northwestern Mediterranean, where sea urchin densities ranged between 0.9 and 3.4 ind m⁻². After 6 months, the cover of non-crustose algae was significantly higher in the plots from which sea urchins had been removed than in control plots (84 vs 67% cover). These removal plots reverted to their original state upon reintroduction of sea urchins. The non-crustose algae consisted of turfing and frondose forms, with the former representing some 70% of the non-crustose algal cover. Change in the cover of turfing algae was responsible for the significant increase in algal development in the sea urchin removal plots. The response of frondose algae to the treatment varied between algal species. It is concluded that grazing by P. lividus exerts a significant effect on habitat structure, even in communities with low sea urchin densities, such as those found in vast areas of the Mediterranean sublittoral.     

The effects of trophic interactions and spatial competition on algal community composition on Hawaiian coral reefs

Much of coral reef ecology has focused on how human impacts change coral reefs to macroalgal reefs. However, macroalgae may not always be a good indicator of reef decline, especially on reefs with significant sea urchin populations, as found in Kenya and Hawaii. This study tests the effects of trophic interactions (i.e. herbivory by fishes and sea urchins) and spatial competition (between algae and coral) on algal community structure of reefs surrounding two Hawaiian Islands that vary in their level of human impacts. Reef‐building organisms (corals and crustose coralline algae) were less abundant and turf algae were more abundant on Maui as compared to Lanai, where human impacts are lower. In contrast to previous studies, we found no evidence that macroalgae increased with human impacts. Instead, low turf and macroalgal abundance were best explained by the interactive effects of coral cover and sea urchin abundance. Fishing sea urchin predators appeared to have cascading effects on the benthic community. The absence of sea urchin predators and high sea urchin densities correspond to a disproportionately high abundance of turf and crustose coralline algae. We propose that high turf algal abundance is a better indicator of reef decline in Hawaii than high macroalgal abundance because turf abundance was highest on reefs with low coral cover and few fish. The results of this study emphasize that understanding changes in community composition are context‐dependent and that not all degraded reefs look the same.     

海参花营养成分分析及其提取物对人卵巢癌细胞雌二醇分泌的影响

海参花是雌海参的性腺,在海参加工过程中常被作为下脚料,但它含有丰富的营养成分和多种活性因子,本研究检测了海参花的一般营养物质、元素含量以及氨基酸组成,同时,研究了海参花水提取物和乙醇提取物对人卵巢癌OVCAR 3细胞增殖以及对其雌二醇分泌的影响。结果表明,海参花含水量为86.63%,粗蛋白、粗脂质和粗多糖（干重）占比分别为65.06%、14.82%和1.93%。共检测出17种氨基酸,总氨基酸含量为519.09 mg/kg。其中,必需氨基酸含量为197.04 mg/kg。海参花还含有多种对人体有益的元素。海参花提取物对人卵巢癌OVCAR 3细胞的增殖有明显的抑制作用,海参花水提取物、乙醇提取物均可提高细胞培养液上清液中雌二醇的含量,其中乙醇提取物可显著提高上清液中雌二醇的含量（P<0.01）。以上表明,海参花的营养丰富,其提取物能显著促进人卵巢癌OVCAR 3细胞雌二醇的分泌调节,因此具有很高的开发利用价值。     

脆嫩–厚成期海带物质成分变化分析研究

采用国家规定的标准方法,分别测定了脆嫩-厚成期(1~5月份)荣成地区养殖海带体内9种物质(蛋白、脂肪、粗纤维、粗灰分、碘、磷、钙、铁和锌)含量,系统分析了各物质成分含量变化规律,以及各成分与海水温度的关联性。分析表明,不同物质质量分数最高值出现时间不同,其中蛋白质和粗灰分质量分数最高值出现在1月份,脂肪、磷和锌出现在2月份,碘和铁出现在3月份,粗纤维和钙出现在5月份。同时,1~5月份各种物质质量分数变化规律也不相同,蛋白质、粗灰分、磷和锌的质量分数呈现下降趋势,粗纤维、钙的质量分数呈上升趋势,而碘和铁的质量分数变化不稳定。蛋白质、脂肪、粗纤维、粗灰分、磷和锌含量变化与温度有显著关联性。     

Dietary Shifts in the Queen Triggerfish, Balistes vetula, in the Absence of its Primary Food Item, Diadema antillarum

Abstract. The long-spined sea urchin Diadema antillarum has been reported to be the major food item of the queen triggerfish, Balistes vetula in the Caribbean. This sea urchin has undergone a massive mortality on reefs throughout the Caribbean during 1983. The dietary habits of twenty-three queen triggerfish from patch reef habitats in Belize were examined. It was found that crabs and chitons now form the major dietary items when D. antillarum is not available. It is suggested that this predator, which was previously specialized on a single food item, is able to shift resource utilization in the absence of its primary food source, and that the queen triggerfish can capture diverse prey from a wide variety of habitats.     

海胆早期血清素能神经系统的发育及功能研究进展

海胆胚胎是研究神经发育生物学的理想生物模型。血清素能神经系统在海胆早期胚胎发育过程中形成最早,主要由血清素能顶端神经节和受体细胞网络构成,对海胆幼虫的游泳行为具有调控功能。本文对海胆幼虫血清素能顶端神经节和受体细胞网络的发育过程,以及血清素能神经系统的功能等方面进行了综述。     

Differences in trophic position among sympatric sea urchin species

Three species of sea urchin regularly co-occur in high abundances on subtidal rocky reefs in south-western Australia. We used two lines of evidence (stable isotope analysis and gut contents analysis), to test whether these species occupy different trophic positions. We looked at five discrete populations to test whether patterns were consistent. The gut contents of Heliocidaris erythrogramma contained almost exclusively fragments of macroalgae, and the δ¹⁵N of muscle was consistent with that expected for a herbivore. In contrast, the gut contents of Phyllacanthus irregularis and Centrostephanus tenuispinus contained a greater proportion of animal tissue, and the δ¹⁵N of muscle suggested that animal tissue was an important source of nutrition. Of the three co-occurring sea urchin species, one (H. erythrogramma) was ecologically dissimilar to the others and occupied a lower trophic position. This pattern was consistent among populations separated by up to 270 km in south-western Australia. Food resource partitioning might be one way in which these species are able to coexist.