title: The environmental quality of Doñana surrounding areas affects the immune transcriptional profile of inhabitant crayfish Procambarus clarkii.
abstract: This study aimed to identify differentially expressed genes in Procambarus clarkii crayfish collected from locations of different environmental qualities in the Doñana National Park surrounding areas. The pollution sustained by the crayfish was confirmed by their hepatopancreatic metal concentration. We generated forward and reverse libraries by suppression subtractive hybridization (SSH) to analyze the transcriptional profiles of crayfish from moderately and highly polluted zones in comparison with the control site within the Doñana Biological Reserve. Forty-three differentially expressed genes were detected, and most of them were identified as genes involved in a variety of biological functions, particularly in the innate immune response. To verify the SSH results and assess interindividual variability nine transcripts (ALP, AST, BTF3, CHIT, CTS, ferritin, HC, HC2, and SPINK4) were selected for absolute quantification by real-time qRT-PCR. The qRT-PCR data revealed substantial differences in the absolute amounts of the nine transcripts and confirmed their up- or down-regulation in the polluted sites. Additionally, a positive and significant linear correlation was found between the hepatopancreatic copper concentration and the levels of the transcripts encoding hemocyanins. Finally, the transcriptomic study was complemented with a detailed analysis of SNP profiles of the selected transcripts that revealed point mutations that might underlie adaptive response to environmental stress in P. clarkii. Overall, this work provides novel insights into the molecular pathways that could mediate the response to environmental pollutants in P. clarkii emphasizing the central role of the immune function and thus, should clearly benefit further immunotoxicological research in this organism.
authors.affiliation: Department of Biochemistry and Molecular Biology, Agrifood Campus of International Excellence (ceiA3), University of Córdoba, Severo Ochoa Building, Rabanales Campus, 14071 Córdoba, Spain.
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Osuna-Jiménez
authors.affiliation: Department of Biochemistry and Molecular Biology, Agrifood Campus of International Excellence (ceiA3), University of Córdoba, Severo Ochoa Building, Rabanales Campus, 14071 Córdoba, Spain.
authors.forename: Nieves
authors.initials: N
authors.lastname: Abril
authors.affiliation: Department of Biochemistry and Molecular Biology, Agrifood Campus of International Excellence (ceiA3), University of Córdoba, Severo Ochoa Building, Rabanales Campus, 14071 Córdoba, Spain.
authors.forename: Amalia
authors.initials: A
authors.lastname: Vioque-Fernández
authors.affiliation: Department of Chemistry and Materials Science, Faculty of Experimental Science, University of Huelva, El Carmen Campus, 21007 Huelva, Spain.
authors.forename: José Luis
authors.initials: JL
authors.lastname: Gómez-Ariza
authors.affiliation: Department of Biochemistry and Molecular Biology, Agrifood Campus of International Excellence (ceiA3), University of Córdoba, Severo Ochoa Building, Rabanales Campus, 14071 Córdoba, Spain. Electronic address: bb2pralm@uco.es.
authors.forename: María-José
authors.initials: MJ
authors.lastname: Prieto-Álamo
authors.affiliation: Department of Biochemistry and Molecular Biology, Agrifood Campus of International Excellence (ceiA3), University of Córdoba, Severo Ochoa Building, Rabanales Campus, 14071 Córdoba, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Pueyo
mesh_terms: D000818:Animals; D060829:Arthropod Proteins / Q000235:genetics* / Q000378:metabolism; D003400:Astacoidea / Q000235:genetics / Q000276:immunology*; D020224:Expressed Sequence Tags; D005786:Gene Expression Regulation; D043143:Hepatopancreas / Q000276:immunology; D008297:Male; D008670:Metals / Q000378:metabolism / Q000633:toxicity*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013030:Spain; D066251:Subtractive Hybridization Techniques; D014874:Water Pollutants, Chemical / Q000378:metabolism / Q000633:toxicity*
chemical_list: D060829:Arthropod Proteins; D008670:Metals; D012333:RNA, Messenger; D014874:Water Pollutants, Chemical
keywords: Absolute transcription profiles; Hepatopancreas; Procambarus clarkii; SNPs; Suppression subtractive hybridization (SSH)