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      Transcriptome analysis on the exoskeleton formation in early developmetal stages and reconstruction scenario in growth-moulting in Litopenaeus vannamei

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          Abstract

          Exoskeleton construction is an important issue in shrimp. To better understand the molecular mechanism of exoskeleton formation, development and reconstruction, the transcriptome of the entire developmental process in Litopenaeus vannamei, including nine early developmental stages and eight adult-moulting stages, was sequenced and analysed using Illumina RNA-seq technology. A total of 117,539 unigenes were obtained, with 41.2% unigenes predicting the full-length coding sequence. Gene Ontology, Clusters of Orthologous Group (COG), the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and functional annotation of all unigenes gave a better understanding of the exoskeleton developmental process in L. vannamei. As a result, more than six hundred unigenes related to exoskeleton development were identified both in the early developmental stages and adult-moulting. A cascade of sequential expression events of exoskeleton-related genes were summarized, including exoskeleton formation, regulation, synthesis, degradation, mineral absorption/reabsorption, calcification and hardening. This new insight on major transcriptional events provide a deep understanding for exoskeleton formation and reconstruction in L. vannamei. In conclusion, this is the first study that characterized the integrated transcriptomic profiles cover the entire exoskeleton development from zygote to adult-moulting in a crustacean, and these findings will serve as significant references for exoskeleton developmental biology and aquaculture research.

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            Structure and mechanical properties of crab exoskeletons.

            The structure and mechanical properties of the exoskeleton (cuticle) of the sheep crab (Loxorhynchus grandis) were investigated. The crab exoskeleton is a natural composite consisting of highly mineralized chitin-protein fibers arranged in a twisted plywood or Bouligand pattern. There is a high density of pore canal tubules in the direction normal to the surface. These tubules have a dual function: to transport ions and nutrition and to stitch the structure together. Tensile tests in the longitudinal and normal to the surface directions were carried out on wet and dry specimens. Samples tested in the longitudinal direction showed a convex shape and no evidence of permanent deformation prior to failure, whereas samples tested in the normal orientation exhibited a concave shape. The results show that the composite is anisotropic in mechanical properties. Microindentation was performed to measure the hardness through the thickness. It was found that the exocuticle (outer layer) is two times harder than the endocuticle (inner layer). Fracture surfaces after testing were observed using scanning electron microscopy; the fracture mechanism is discussed.
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              Insect cuticular proteins.

              Insect cuticles are composite structural materials with mechanical properties optimal for their biological functions. The bulk properties of cuticles are to a large extent determined by the interactions between the various components, mainly the chitin filament system and the proteins. The various cuticular types show pronounced differences in mechanical properties, and it is suggested that these differences can be related to the properties of the individual proteins and to the degree of secondary stabilization (sclerotization). The amino acid sequences, which have been obtained for insect cuticular proteins either by direct sequencing of purified proteins or by deduction from corresponding DNA-sequences, are listed according to insect order and species. Extensive sequence similarity is observed among several cuticular proteins obtained from different insect orders. Other cuticular proteins are characterized by repeated occurrence of a few small motifs consisting mainly of hydrophobic residues. The latter group of proteins has so far only been reported from stiff cuticles. The possible relevance of the various motifs and repeats for protein interaction and the mechanical properties of cuticles is discussed.
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                Author and article information

                Contributors
                xjzhang@qdio.ac.cn
                jhxiang@qdio.ac.cn
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                24 April 2017
                24 April 2017
                2017
                : 7
                : 1098
                Affiliations
                [1 ]ISNI 0000000119573309, GRID grid.9227.e, Key Laboratory of Experimental Marine Biology, Institute of Oceanology, , Chinese Academy of Sciences, ; Qingdao, 266071 China
                [2 ]Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071 China
                [3 ]ISNI 0000 0001 2152 3263, GRID grid.4422.0, , Ocean University of China, ; Qingdao, 266071 China
                Article
                1220
                10.1038/s41598-017-01220-6
                5430884
                28439089
                b690fa77-5df4-4bbb-9d02-17a3203ca424
                © The Author(s) 2017

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 8 December 2016
                : 21 March 2017
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