Gene expression profile analysis of Manila clam (Ruditapes philippinarum) hemocytes after a Vibrio alginolyticus challenge using an immune-enriched oligo-microarray

详细信息    查看全文

• 作者：Rebeca Moreira ; Massimo Milan ; Pablo Balseiro ; Alejandro Romero…
• 关键词：Ruditapes philippinarum ; Vibrio alginolyticus ; Hemocytes ; Oligo ; microarray ; Gene ontology ; Blast2GO ; Immune response
• 刊名：BMC Genomics
• 出版年：2014
• 出版时间：December 2014
• 年：2014
• 卷：15
• 期：1
• 全文大小：950 KB
• 参考文献：1. Gestal, C, Roch, P, Renault, T, Pallavicini, A, Paillard, C, Novoa, B, Oubella, R, Venier, P, Figueras, A (2008) Study of diseases and the immune system of bivalves using molecular biology and genomics. Rev Fish Sci 16: pp. 131-154 CrossRef
  2. Paillard, C, Leroux, F, Borrego, JJ (2004) Bacterial disease in marine bivalves, Review of recent studies. Trends and evolution. Aquat Living Resour 17: pp. 477-498 CrossRef
  3. Villalba, A, Reece, KS, Ordás, MC, Casas, SM, Figueras, A (2004) Perkinsosis in molluscs: A review. Aquat Living Resour 17: pp. 411-432 CrossRef
  4. Waki, T, Shimokawa, J, Watanabe, S, Yoshinaga, T, Ogawa, K (2012) Experimental challenges of wild Manila clams with Perkinsus species isolated from naturally infected wild Manila clams. J Invertebr Pathol 111: pp. 50-55 CrossRef
  5. Allam, B, Paillard, C, Auffret, M (2000) Alterations in hemolymph and extrapallial fluid parameters in the Manila clam, Ruditapes philippinarum, challenged with the pathogen Vibrio tapetis. J Invertebr Pathol 76: pp. 63-69 CrossRef
  6. Gómez-León, J, Villamil, L, Lemos, ML, Novoa, B, Figueras, A (2005) Isolation of V. alginolyticus and Vibrio splendidus from aquacultured carpet shell clam (Ruditapes decussatus) larvae associated with mass mortalities. Appl Environ Microb 71: pp. 98-104 CrossRef
  7. Canesi, L, Gallo, G, Gavioli, M, Pruzzo, C (2002) Bacteria–hemocyte interactions and phagocytosis in bivalves. Microsc Res Technol 57: pp. 469-476 CrossRef
  8. Olafsen, JA (1995) Role of lectins (C-reactive protein) in defense of marine bivalves against bacteria. Adv Exp Med Biol 371A: pp. 343-348 CrossRef
  9. Ordás, MC, Novoa, B, Figueras, A (2000) Modulation of the chemiluminescence response of Mediterranean mussel (Mytilus galloprovincialis) haemocytes. Fish Shellfish Immunol 10: pp. 611-622 CrossRef
  10. Ordás, MC, Ordás, A, Beloso, C, Figueras, A (2000) Immune parameters in carpet shell clams naturally infected with Perkinsus atlanticus. Fish Shellfish Immunol 10: pp. 597-609 CrossRef
  11. Tafalla, C, Gómez-León, J, Novoa, B, Figueras, A (2003) Nitric oxide production by carpet shell clam (Ruditapes decussatus) hemocytes. Dev Comp Immunol 27: pp. 197-205 CrossRef
  12. Bayne, CJ, Sminia, T, Van der Knaap, WPW Immunological Memory: Status of Molluscan Studies. In: Manning, MJ eds. (1980) Phylogeny of Immunological Memory. Elsevier, Amsterdam, pp. 57-64
  13. Gueguen, Y, Cadoret, JP, Flament, D, Barreau-Roumiguiere, C, Girardot, AL, Garnier, J, Hoareau, A, Bachere, E, Escoubas, JM (2003) Immune gene discovery by expressed sequence tags generated from hemocytes of the bacteria-challenged oyster, Crassostrea gigas. Gene 303: pp. 139-145 CrossRef
  14. Costa, MM, Novoa, B, Figueras, A (2008) Influence of β-glucans on the immune responses of carpet shell clam (Ruditapes decussatus) and Mediterranean mussel (Mytilus galloprovincialis). Fish Shellfish Immunol 24: pp. 498-505 CrossRef
  15. Costa, MM, Prado-Alvarez, M, Gestal, C, Roch, P, Li, H, Novoa, B, Figueras, A (2009) Functional and molecular immune response of Mediterranean mussel (Mytilus galloprovincialis) hemocytes against pathogen associated molecular patterns and bacteria. Fish Shellfish Immunol 26: pp. 515-523 CrossRef
  16.
• 刊物主题：Life Sciences, general; Microarrays; Proteomics; Animal Genetics and Genomics; Microbial Genetics and Genomics; Plant Genetics & Genomics;
• 出版者：BioMed Central
• ISSN：1471-2164

文摘

Background The Manila clam (Ruditapes philippinarum) is a cultured bivalve with worldwide commercial importance, and diseases cause high economic losses. For this reason, interest in the immune genes in this species has recently increased. The present work describes the construction of the first R. philippinarum microarray containing immune-related hemocyte sequences and its application to study the gene transcription profiles of hemocytes from clams infected with V. alginolyticus through a time course. Results The complete set of sequences from R. philippinarum available in the public databases and the hemocyte sequences enriched in immune transcripts were assembled successfully. A total of 12,156 annotated sequences were used to construct the 8 ×15 k oligo-microarray. The microarray experiments yielded a total of 579 differentially expressed transcripts. Using the gene expression results, the associated Gene Ontology terms and the enrichment analysis, we found different response mechanisms throughout the experiment. Genes related to signaling, transcription and apoptosis, such as IL-17D, NF-κB or calmodulin, were typically expressed as early as 3 hours post-challenge (hpc), while characteristic immune genes, such as PGRPs, FREPs and defense proteins appeared later at 8 hpc. This immune-triggering response could have affected a high number of processes that seemed to be activated 24 hpc to overcome the Vibrio challenge, including the expression of many cytoskeleton molecules, which is indicative of the active movement of hemocytes. In fact functional studies showed an increment in apoptosis, necrosis or cell migration after the infection. Finally, 72 hpc, activity returned to normal levels, and more than 50% of the genes were downregulated in a negative feedback of all of the previously active processes. Conclusions Using a new version of the R. philippinarum oligo-microarray, a putative timing for the response against a Vibrio infection was established. The key point to overcome the challenge seemed to be 8 hours after the challenge, when we detected immune functions that could lead to the destruction of the pathogen and the activation of a wide variety of processes related to homeostasis and defense. These results highlight the importance of a fast response in bivalves and the effectiveness of their innate immune system.