Changes in the Organics Metabolism in the Hepatopancreas Induced by Eyestalk Ablation of the Chinese Mitten Crab Eriocheir sinensis Determined via Transcriptome and DGE Analysis

PLOS ONE, Apr 2014

Background To understand the regulation mechanism of eyestalk ablation on the activities of hepatopancreas, Illumina RNA-Seq and digital gene expression (DGE) analyses were performed to investigate the transcriptome of the eyestalk, Y-organ, and hepatopancreas of E. sinensis and to identify the genes associated with the hepatopancreas metabolism that are differentially expressed under eyestalk ablation conditions. Results A total of 58,582 unigenes were constructed from 157,168 contigs with SOAPdenovo. A BlastX search against the NCBI Nr database identified 21,678 unigenes with an E-value higher than 10−5. Using the BLAST2Go and BlastAll software programs, 6,883 unigenes (11.75% of the total) were annotated to the Gene Ontology (GO) database, 7,386 (12.6%) unigenes were classified into 25 Clusters of Orthologous Groups of Proteins (COGs), 16,200 (27.7%) unigenes were assigned to 242 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and1,846 unigenes were matched to “metabolism pathways”. The DGE analysis revealed that 1,416 unigenes were significantly differentially expressed in the hepatopancreas, of which 890 unigenes were up-regulated and 526 unigenes were down-regulated. Of the differentially expressed genes, 382 unigenes were annotated and 63 were classified into metabolism pathways. The results of the real-time polymerase chain reaction (PCR) analysis of four unigenes related to carbohydrate metabolism were consistent with those obtained from the DGE analysis, which demonstrates that the sequencing data were satisfactory for further gene expression analyses. Conclusion This paper reported the transcriptom of the eyestalk, Y-organ, and hepatopancreas from E. sinensis. DGE analysis provided the different expressed genes of the metabolism processes in hepatopancreas that are affected by eyestalk ablation. These findings will facilitate further investigations on the mechanisms of the metabolism of organic substances during development and reproduction in crustaceans.

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Changes in the Organics Metabolism in the Hepatopancreas Induced by Eyestalk Ablation of the Chinese Mitten Crab Eriocheir sinensis Determined via Transcriptome and DGE Analysis

et al. (2014) Changes in the Organics Metabolism in the Hepatopancreas Induced by Eyestalk Ablation of the Chinese Mitten Crab Eriocheir sinensis Determined via Transcriptome and DGE Analysis. PLoS ONE 9(4): e95827. doi:10.1371/journal.pone.0095827 Editor: Senjie Lin Changes in the Organics Metabolism in the Hepatopancreas Induced by Eyestalk Ablation of the Chinese Mitten Crab Eriocheir sinensis Determined via Transcriptome and DGE Analysis Yan Sun 0 Yichen Zhang 0 Yichen Liu 0 Shuxia Xue 0 Xuyun Geng 0 Tong Hao 0 Jinsheng Sun 0 0 1 Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal University , Tianjin , People's Republic of China, 2 Tianjin Center for Control and Prevention of Aquatic Animal Infectious Disease , Tianjin , People's Republic of China Background: To understand the regulation mechanism of eyestalk ablation on the activities of hepatopancreas, Illumina RNA-Seq and digital gene expression (DGE) analyses were performed to investigate the transcriptome of the eyestalk, Yorgan, and hepatopancreas of E. sinensis and to identify the genes associated with the hepatopancreas metabolism that are differentially expressed under eyestalk ablation conditions. Results: A total of 58,582 unigenes were constructed from 157,168 contigs with SOAPdenovo. A BlastX search against the NCBI Nr database identified 21,678 unigenes with an E-value higher than 1025. Using the BLAST2Go and BlastAll software programs, 6,883 unigenes (11.75% of the total) were annotated to the Gene Ontology (GO) database, 7,386 (12.6%) unigenes were classified into 25 Clusters of Orthologous Groups of Proteins (COGs), 16,200 (27.7%) unigenes were assigned to 242 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and1,846 unigenes were matched to ''metabolism pathways''. The DGE analysis revealed that 1,416 unigenes were significantly differentially expressed in the hepatopancreas, of which 890 unigenes were up-regulated and 526 unigenes were down-regulated. Of the differentially expressed genes, 382 unigenes were annotated and 63 were classified into metabolism pathways. The results of the real-time polymerase chain reaction (PCR) analysis of four unigenes related to carbohydrate metabolism were consistent with those obtained from the DGE analysis, which demonstrates that the sequencing data were satisfactory for further gene expression analyses. Conclusion: This paper reported the transcriptom of the eyestalk, Y-organ, and hepatopancreas from E. sinensis. DGE analysis provided the different expressed genes of the metabolism processes in hepatopancreas that are affected by eyestalk ablation. These findings will facilitate further investigations on the mechanisms of the metabolism of organic substances during development and reproduction in crustaceans. - Funding: This work was financially supported by grants from the National High-Tech Research and Development Program of China (863 programs, 2012A A10A401 and 2012AA092205); the National Key techno&loDgy PrRogram (2012BAD26B04); the program from key laboratory of freshwater aquatic genetic resources certificated by ministry of agriculture and the Major State Basic Research Development Program of China (973 programs, 2012CB114405). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. The hepatopancreas of crustaceans was described as a metabolic factory, which was the center of lipid and carbohydrate metabolism [1]. The functions of the hepatopancreas are considered to include the storage and depletion of organic substances to support various significant life activities, such as ovarian maturation, vitellogenin synthesis and molting [24]. Murugesan [5] reported the effect of eyestalk ablation on the biochemical composition of the crab, Charybdis lucifera, the result showed that the content of carbohydrate, lipid, fatty acids and protein in crabs of eyestalk ablation were higher than intact crabs, eyestalk ablation can cause the glycogen synthesis [6] in hepatopancreas. However, the exact regulation mechanism of eyestalk ablation on the activities of hepatopancreas is not clear. Some studies revealed that when the hepatopancreas was incubated with CHH; the rate of glucose into glycogen was decreased [7]. These results suggest a physiological role for CHH in the control of carbohydrates and lipid metabolism in crustacean. Crustacean hyperglycemic hormone (CHH), a neuropeptide synthesized and secreted from the X-organ/sinus gland (XO/SG) complex in the eyestalk, plays a central role in the regulation of energy metabolism [8]. The hepatopancreas is the primary tissue on which CHH act [9]. A number of studies have reported that CHH has the function to elevate the glucose level in the hemolymph of many decapods [1012]. Besides the researches on the macro level, some genes related to gly (...truncated)


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Yan Sun, Yichen Zhang, Yichen Liu, Shuxia Xue, Xuyun Geng, Tong Hao, Jinsheng Sun. Changes in the Organics Metabolism in the Hepatopancreas Induced by Eyestalk Ablation of the Chinese Mitten Crab Eriocheir sinensis Determined via Transcriptome and DGE Analysis, PLOS ONE, 2014, Volume 9, Issue 4, DOI: 10.1371/journal.pone.0095827