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Fisheries Science

, Volume 85, Issue 6, pp 979–989 | Cite as

Comparative transcriptome analysis reveals the expression and characterization of digestive enzyme genes in the hepatopancreas of the Chinese mitten crab

  • Huayun Guo
  • Dan Tang
  • Xueling Shi
  • Qiong Wu
  • Ruobing Liu
  • Boping Tang
  • Zhengfei WangEmail author
Original Article Biology

Abstract

The Chinese mitten crab Eriocheir japonica sinensis is one of the most common aquaculture species cultivated in China. The crab is an omnivore, and its hepatopancreas absorbs and stores nutrients. The aim of this study was to elucidate the expressions of the digestive enzyme genes and determine their respective roles in regulating digestive capacity in E. j. sinensis. We sequenced the hepatopancreatic transcriptomes of crabs fed a meat diet (MD), a vegetarian diet (VD), or a mixed diet (MV) and compared the gene expression patterns of these three groups of crabs. A total of 305,887 unigenes were obtained, of which 8747, 10,963, and 8877 were significantly differentially expressed in the comparisons between the MD and MV, VD and MV, and MD and VD diets, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) database-based enrichment analysis revealed that the differentially expressed gene (DEG) responses in the hepatopancreases to the MD mainly involved the “pancreatic secretion,” “glutathione metabolism,” “sphingolipid metabolism,” “fatty acid metabolism,” and “glycerolipid metabolism pathways.” DEG responses to the VD based on KEGG analysis mainly involved the “galactose metabolism,” “starch and sucrose metabolism,” and “fructose and mannose metabolism” pathways. The key digestive enzymes, including trypsin, β-glucosidase, chitinase, and triacylglycerol lipase, were identified. Our results further our understanding of crustacean hepatopancreatic functions during food digestion and provide resources for further studies regarding the molecular basis of omnivorous diets in crustaceans.

Keywords

Transcriptome Chinese mitten crab Hepatopancreas Omnivore Digestive enzyme 

Notes

Acknowledgements

This study was funded by the National Natural Science Foundation of China (Grant Number 31702014), and Doctoral Scientific Research Foundation of Yancheng Teachers University to ZFW, and Open Foundation of Jiangsu Key Laboratory for Bioresources of Saline Soils (Grant Number JKLBS2016007).

Author contributions

HYG, DT, XLS, WQ, RBL, BPT, and ZFW designed and conceived the experiment. ZFW and HYG performed the data analysis and drafted the manuscript. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare there are no competing interests.

Ethical approval

The sampling location was not privately owned or protected, and field sampling did not involve protected species.

Supplementary material

12562_2019_1358_MOESM1_ESM.pdf (201 kb)
Online Resource 1. Start and end body weights of Eriocheir japonica sinensis. (PDF 201 kb)
12562_2019_1358_MOESM2_ESM.pdf (201 kb)
Online Resource 2. Carbohydrate, lipid, and protein ratios in MD, VD and MV. (PDF 201 kb)
12562_2019_1358_MOESM3_ESM.pdf (251 kb)
Online Resource 3. Species distribution of the BLASTx matches of the hepatopancreas transcriptome unigenes. (PDF 250 kb)
12562_2019_1358_MOESM4_ESM.pdf (292 kb)
Online Resource 4. GO function classifcation of the assembled unigenes in the hepatopancreas of Eriocheir japonica sinensis, including three classes of biological process, cellular process and molecular function. (PDF 291 kb)
12562_2019_1358_MOESM5_ESM.pdf (467 kb)
Online Resource 5. Significantly enriched pathways in MD vs MV, VD vs MV and MD vs VD under KEGG enrichment analysis. (PDF 466 kb)
12562_2019_1358_MOESM6_ESM.pdf (360 kb)
Online Resource 6. DEGs numbers in KEGG enrichment terms involved in nutrition metabolism. (PDF 360 kb)
12562_2019_1358_MOESM7_ESM.pdf (73 kb)
Online Resource 7. Phylogenetic tree of the chitinase family. (PDF 73 kb)
12562_2019_1358_MOESM8_ESM.pdf (206 kb)
Online Resource 8. Gene expression patterns of different digestive enzymes in Eriocheir japonica sinensis fed with three diets (MD, VD and MV). (PDF 205 kb)

References

  1. Birk RZ, Brannon PM (2004) Regulation of pancreatic lipase by dietary medium chain triglycerides in the weanling rat. Pediatr Res 55:921–926CrossRefGoogle Scholar
  2. Bui THH, Lee SY (2015) Endogenous cellulase production in the leaf litter foraging mangrove crab Parasesarma erythodactyla. Comp Biochem Physiol B Biochem Mol Biol 179:27–36CrossRefGoogle Scholar
  3. Cannicci S, Schubart CD, Innocenti G et al (2017) A new species of the genus Parasesarma De Man 1895 from East African mangroves and evidence for mitochondrial introgression in sesarmid crabs. Zool Anz 269:89–99CrossRefGoogle Scholar
  4. Chu Y, Corey DR (2012) RNA sequencing: platform selection, experimental design, and data interpretation. Nucleic Acid Ther 22:271–274CrossRefGoogle Scholar
  5. Dammannagoda LK, Pavasovic A, Prentis PJ et al (2015) Expression and characterization of digestive enzyme genes from hepatopancreatic transcripts from redclaw crayfish (Cherax quadricarinatus). Aquac Nutr 21:868–880CrossRefGoogle Scholar
  6. Dittel AI, Epifanio CE (2009) Invasion biology of the Chinese mitten crab Eriochier sinensis: a brief review. J Exp Mar Bio Ecol 374:79–92CrossRefGoogle Scholar
  7. Ghosh D, Porter E, Shen B et al (2002) Paneth cell trypsin is the processing enzyme for humandefensin-5. Nat Immunol 3:583–590CrossRefGoogle Scholar
  8. Gross PS, Bartlett TC, Browdy CL et al (2001) Immune gene discovery by expressed sequence tag analysis of hemocytes and hepatopancreas in the Pacific White Shrimp, Litopenaeus vannamei, and the Atlantic White Shrimp L. setiferus. Dev Comp Immunol 25:565–577CrossRefGoogle Scholar
  9. Huang S, Wang J, Yue W et al (2015) Transcriptomic variation of hepatopancreas reveals the energy metabolism and biological processes associated with molting in Chinese mitten crab Eriocheir sinensis. Sci Rep 5:14015CrossRefGoogle Scholar
  10. Jiang H, Yin Y, Zhang X et al (2009) Chasing relationships between nutrition and reproduction: a comparative transcriptome analysis of hepatopancreas and testis from Eriocheir sinensis. Comp Biochem Physiol Part D Genomics Proteomics 4:227–234CrossRefGoogle Scholar
  11. Jiankai W, Xiaojun Z, Yang Y et al (2014) Comparative transcriptomic characterization of the early development in Pacific white shrimp Litopenaeus vannamei. PLoS ONE 9:e106201CrossRefGoogle Scholar
  12. Li X, Cui Z, Liu Y et al (2013) Transcriptome analysis and discovery of genes involved in immune pathways from hepatopancreas of microbial challenged mitten crab Eriocheir sinensis. PLoS ONE 8:e68233CrossRefGoogle Scholar
  13. Li X, Xu Z, Zhou G et al (2015a) Molecular characterization and expression analysis of five chitinases associated with molting in the Chinese mitten crab, Eriocheir sinensis. Comp Biochem Physiol B Biochem Mol Biol 187:110–120CrossRefGoogle Scholar
  14. Li Y, Hui M, Cui Z (2015b) Comparative transcriptomic analysis provides insights into the molecular basis of the metamorphosis and nutrition metabolism change from zoea to megalopae in Eriocheir sinensis. Comp Biochem Physiol Part D Genomics Proteomics 13:1–9CrossRefGoogle Scholar
  15. Merzendorfer H (2013) Insect-derived chitinases. Adv Biochem Eng Biotechnol 136:19–50PubMedGoogle Scholar
  16. Michiels MS, Valle JCD, Mañanes AAL (2017) Trypsin and N-aminopeptidase (APN) activities in the hepatopancreas of an intertidal euryhaline crab: biochemical characteristics and differential modulation by histamine and salinity. Comp Biochem Physiol A Mol Integr Physiol 204:228–235CrossRefGoogle Scholar
  17. Mykles DL (2011) Ecdysteroid metabolism in crustaceans. J Steroid Biochem Mol Biol 127:196–203CrossRefGoogle Scholar
  18. R Development Core Team (2015) R: A language and environment for statistical computing. R Foundation for Statistical Computing, ViennaGoogle Scholar
  19. Rao R, Zhu YB, Alinejad T et al (2015) RNA-Seq analysis of Macrobrachium rosenbergii hepatopancreas in response to Vibrio parahaemolyticus infection. Gut Pathog 7:6CrossRefGoogle Scholar
  20. Rivera-Pérez C, García-Carreño F (2011) Effect of fasting on digestive gland lipase transcripts expression in Penaeus vannamei. Mar Genom 4:273–278CrossRefGoogle Scholar
  21. Rojo L, Garcíacarreño F (2012) Cold-adapted digestive aspartic protease of the clawed lobsters Homarus americanus and Homarus gammarus: biochemical characterization. Mar Biotechnol 15:87–96CrossRefGoogle Scholar
  22. Roux MM, Pain A, Klimpel KR et al (2002) The lipopolysaccharide and beta-1,3-glucan binding protein gene is upregulated in white spot virus-infected shrimp (Penaeus stylirostris). J Virol 76:7140–7149CrossRefGoogle Scholar
  23. Rudnick DA, Hieb K, Grimmer KF et al (2003) Patterns and processes of biological invasion: the Chinese mitten crab in San Francisco Bay. Basic Appl Ecol 4:249–262CrossRefGoogle Scholar
  24. Salma U, Uddowla MH, Kim M et al (2012) Five hepatopancreatic and one epidermal chitinases from a pandalid shrimp (Pandalopsis japonica): cloning and effects of eyestalk ablation on gene expression. Comp Biochem Physiol B Biochem Mol Biol 161:197–207CrossRefGoogle Scholar
  25. Shi Y, Burn P (2004) Lipid metabolic enzymes: emerging drug targets for the treatment of obesity. Nat Rev Drug Discov 3:695–710CrossRefGoogle Scholar
  26. Wang L, Yan B, Liu N et al (2009) Effects of cadmium on glutathione synthesis in hepatopancreas of fresh water crab, Sinopotamon yangtsekiense. Chemosphere 74:51–56CrossRefGoogle Scholar
  27. Wang W, Wu X, Liu Z et al (2014) Insights into hepatopancreatic functions for nutrition metabolism and ovarian development in the crab Portunus trituberculatus: gene discovery in the comparative transcriptome of different hepatopancreas stages. PLoS ONE 9:e84921CrossRefGoogle Scholar
  28. Wang Z, Xu S, Du K et al (2016) Evolution of digestive enzymes and RNASE1 provides insights into dietary switch of cetaceans. Mol Biol Evol 33:3144–3157CrossRefGoogle Scholar
  29. Wang Z, Bai Y, Zhang D et al (2018) Adaptive evolution of osmoregulatory-related genes provides insight into salinity adaptation in Chinese mitten crab, Eriocheir sinensis. Genetica 146:303–311CrossRefGoogle Scholar
  30. Watanabe H, Tokuda G (2010) Cellulolytic systems in insects. Annu Rev Entomol 55:609–632CrossRefGoogle Scholar
  31. Wei W, Wu X, Liu Z et al (2014) Insights into hepatopancreatic functions for nutrition metabolism and ovarian development in the crab Portunus trituberculatus: gene discovery in the comparative transcriptome of different hepatopancreas stages. PLoS ONE 9:e84921CrossRefGoogle Scholar
  32. Wei B, Yang Z, Wang J et al (2017) Effects of dietary lipids on the hepatopancreas transcriptome of Chinese mitten crab (Eriocheir sinensis). PLoS ONE 12:e182087Google Scholar
  33. Zhu B, Tang L, Yu Y et al (2017) Identification of ecdysteroid receptor-mediated signaling pathways in the hepatopancreas of the red swamp crayfish, Procambarus clarkii. Gen Comp Endocrinol 246:372–381CrossRefGoogle Scholar

Copyright information

© Japanese Society of Fisheries Science 2019

Authors and Affiliations

  1. 1.Jiangsu Key Laboratory for Bioresources of Saline Soils, Jiangsu Synthetic Innovation Center for Coastal Bio-agriculture, Jiangsu Provincial Key Laboratory of Coastal Wetland Bioresources and Environmental Protection, School of Ocean and Biological EngineeringYancheng Teachers UniversityYanchengChina
  2. 2.College of Biotechnology and Pharmaceutical EngineeringNanjing Tech UniversityNanjingChina

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