Ontogenesis of main organs and cloning and expression of two immune-related genes in Litopenaeus van

来源 :中国海洋大学 | 被引量 : 0次 | 上传用户:ontheroad2005
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
White leg shrimp Litopenaeus vannamei is a prime aquaculture species, and it covers 90%of western hemisphere culture and is also extensively farmed in Asia. The estimated annual production from Asia during 2002 was 316000 mt. In China, exotic species (L. vannamei) had introduced during 1988 for experimental purposes and by 1998 is being successfully cultured in commercial scale; over 1000 hatcheries are in operation. The 300000 tons and 210000 tons harvested during 2003 in marine and freshwater farming respectively. To give more understanding for the white leg shrimp, we have investigated the genesis of important systems and organ, including digestive system, nervous system and haematopoetic tissue, furthermore dealed with two important immune related genes.  Our investigation on ontogenesis of digestive system in Litopenaeus vannamei, revealed that in non-feeding stages the yolk mass is the indigenous source of food and until the fifth nauplius (N5) stage retained in foregut cavity, absorption is gradual in nauplius (N) stages while disappearance occurs at N6 stage. The anterior midgut caecum generates at N3 and it reaches maximum size in Z3; thereafter in successive stages it reduces in size and becomes a single structure in post-larval stage (P3-P4), named the anterior diverticulum. The lateral midgut caecum first appears at N6 stage and lengthens along both sides of the midgut and then ramifies to form lobes as larval development. Finally, these lobes become a compact structure, the hepatopancreas. The midgut lumen is quite apparent in N stages and only becomes wider in successive stages; it is without a cuticle lining and there is no difference among all investigated stages. The foregut in N stages is a simple tube and pyloric and cardiac chamber occurs in the third mysis (M3) stage. In the cardiac chamber of post-larval stages, folds were noted and number of that increased gradually. The gastric mill is absent in all investigated larval stages (P1-P10). The pyloric chamber is smaller than the cardiac chamber and possesses a gland filter, which first appears in M3 stage and forms a more complete shape in forthcoming stages. The hindgut gland first appears in P2 and in successive stages it shows more protuberance and complexity. The hindgut or rectum is a simple tube. Cells of the digestive system in all feeding stages show the presence of acid mucins and general protein, while the inner margins of the foregut and connective tissues of the gland filter are positive for collegen.  The nervous system plays pivotal role in co-ordination of body activities and coping with the task of controlling the behavioral activities during larval development. In present study, we give a basic description in the early stage morphology of nervous system in L. vannamei by classical histological techniques. Results revealed that neuropil first emerged in N1 stage and the front part of nervous system started getting structural shape in N3 where the ganglions were coalesced in front part of head. The complete structure of central nervous system including proto-, deuto-and tritocerebrum was observed in N6 stage. The structural expansion of nervous system in successive stages was continued and protocerebrum differentiated into anterior and posterior median protocerebrum at Z3 stage. The optic ganglions, sub-esophageal ganglions and pleon ganglions were distinct in zoea stage. Furthermore, we discuss the relativity between the development of olfactory lobes and the voluntary movement of larva.  It is believed that haematopoietic tissue (HPT) is the source of haemocytes production in crustaceans and defensive role of haemocytes is progressing but investigations on structure of HPT are limited hence this study has been designed in which we investigated the genesis and morphology of haematopoietic tissue (HPT) and types of haemocyte based on its morphology and cytochemical feature using histological and histochemistical methods. The results revealed that HPT in L. vannamei is not visible histologically in mysis and early two post larvae (P1 & P2), and emerges in P3 which locates in epigastric region. The HPT showed a continuous propagation of cells in successive post larval stages. Four types of cells can be identified in the HPT from adult specimen and mitotic activity is visible on occasion in the HPT. The haemocytes has been differentiated into five types and some of these types are co-related with the HPT cells. The cytochemical studies suggested that type Ⅰand type Ⅱ cells of haemocytes are PAS positive while scanty presence of prophenoloxidase was observed in type Ⅰ cells similarly the type Ⅰ cells are Sudan Black B positive and rest of the cells showed weak activity against the lipid detective stain. Best of knowledge this is the first effort towards the genesis of HPT and its relation with circulating haemocytes in L. vannamei.  The second part of the study focuses two important immune related genes Cyclophilin A and Proliferating cell nuclear antigen (PCNA); the obtained results are as below respectively.  Cyclophilins are ubiquitous proteins with an enzymatic activity of peptidyl-prolyl cis-trans isomerase (PPlase), which play important roles in a variety of stress responses. In present investigation a Cyclophilin A gene has been cloned from white leg shrimp (Litopenaeus vannamei) by rapid amplification of cDNA (RACE) and anchored PCR method. The full length of the shrimp L. vannamei CypA cDNA (LvCypA) is 855 bp, containing 495 bp of open reading frame (ORF), encoding 164 amino acids with an estimated molecular mass of 17.6 kDa. The LvCypA has high homology with that of other known organisms and it has four β-strands. The real time PCR analysis revealed that its expression is ubiquitous and expression level is higher in muscle, gill, lymphoid organ and hepatopancrease respectively. The tissues challenged with Vibrio angullarium revealed that in hepatopancrease it was the lowest at 6 h and highest in 48 h. The results revealed that the LvCypA might involve in potential role of innate immunity.  The proliferating cell nuclear antigen (PCNA) has important role in nucleic acid metabolism and its immune responses have also been reported. In present investigation the PCNA was cloned from L. vannamei (LvPCNA). The full length cDNA sequence of LvPCNA encodes 261 amino acids showing high identity with PCNAs reported in other species. Tissue distribution of the LvPCNA revealed that its expression is obvious in gill and muscle. In order to understand the response of bacterial challenge, mRNA level of LvPCNA in hepatopancrease, was analyzed after challenged by V. anguillarum. The obtained expression profile of LvPCNA in tissue is variable in different time point. The data enhance the existing knowledge of defense mechanisms of shrimp against bacteria.
其他文献
过去50年间,在全球变化和人为作用不断加强的背景下,莱州湾南岸地理环境退化严重,主要表现为气候变化、海岸侵蚀、海水入侵、环境质量和渔业资源下降以及滨海湿地退化。本文在对大量实测资料和前人研究工作进行分析总结的基础上,对莱州湾南岸地理环境退化的过程、幅度、原因、趋势及其相互之间的关系进行了系统研究,提出了莱州湾南岸资源环境可持续发展管理对策与建议。得到了如下认识: (1)受全球气候变化影响,近
稀有鮈鲫(Gobiocypris rarus)是我国特有的一种小型鲤科鱼类,并已作为一种新型的模式实验鱼应用于鱼病学、遗传学、环境科学等多个研究领域。其所具有的优良特性使其具备通过
学位
目的:血栓栓塞性疾病(thrombotic disease,TD)是一种常见病和多发病,发病率、致残率、致死率皆高。药物溶栓是目前临床最有效的治疗手段。迄今为止,溶栓剂已发展了三代,但仍
2013年被国外媒体称为“大数据元年”。大数据如浪潮一般席卷全世界,为社会的发展带来大量商机,也成为影响媒体发展的一股重要力量。在媒体广告运营中,大数据的运用,特别是用
8月14日,由中国商务部和尼泊尔商业部联合主办的“中国—尼泊尔贸易项目签约仪式”14日上午在加德满都举行。两国企业共签署了33项贸易合同,合同金额共计2116.5万美元,相当于
特点测量范围大150mm-1200mm对轴向工件进行实时测量,适用光滑或断续表面的工件安装灵活方便,无需特殊校准,对标准件简单调零即可可靠,坚固,密封性好直径轮廓测量工件直径超
提出了一种新的解决稀疏支持向量机的方法-基于经典增广拉格朗日框架的交替线性化法.数值结果证明我们方法的有效性. A new method of solving sparse support vector machi
水产品中富含天然动物蛋白和多种维生素,日益受到人们的重视与青睐,水产养殖已成为各国满足高品质水产品产量的重要手段。我国水产养殖年产量已达4000多万吨,虽是渔业大国,但
本文以朱夏先生活动论构造历史观为指导,利用横穿苏北盆地和南黄海地区的36条长约8000km地震剖面、4口测井资料以及86口钻井分层资料、结合前人的野外露头研究资料、成果和重
本文不长,其中的信息却极为重要,请你细细品读,对于从事中医药项目的创业者来说,这样的商机弥足珍贵。关于高血压,我想大家应该都比较熟悉了,目前我国每10人中有近3人为高血