title: Site-related differences in gene expression and bacterial densities in the mussel Bathymodiolus azoricus from the Menez Gwen and Lucky Strike deep-sea hydrothermal vent sites.
abstract: The deep-sea hydrothermal vent mussel Bathymodiolus azoricus is a symbiont bearing bivalve that is found in great abundance at the Menez Gwen and Lucky Strike hydrothermal vent sites and in close vicinity of the Azores region near the Mid-Atlantic Ridge (MAR). The physiological relationships that vent mussels have developed with their physical and chemical environments are likely to influence global gene expression profiles providing thus the means to investigate distinct biological markers predicting the origin of Bathymodiolus sp. irrespectively of their geographical localization. Differences found at gene expression levels, and between fluorescence in situ hybridization (FISH) and 16S rRNA amplicon sequencing results provided experimental evidence for the distinction of both Menez Gwen and Lucky Strike vent mussel individuals based on bacterial and vent mussel gene expression signatures and on the constitutive distribution and relative abundance of endosymbiotic bacteria within gill tissues. Our results confirmed the presence of methanotroph endosymbionts in Menez Gwen vent mussels whereas Lucky Strike specimens seem to harbor a different bacterial morphotype when a methane monooxygenase gene specific probe was used. No qualitative differences could be visualized between Menez Gwen and Lucky Strike individuals when tested with a sulfur-oxidizing-related probe. Quantitative PCR (qPCR) studies revealed different gene expression profiles in both Menez Gwen and Lucky Strike mussel gill tissues for the immune genes selected. Genes encoding transcription factors presented noticeably low levels of fold expression whether in Menez Gwen or Lucky Strike animals whereas the genes encoding effector molecules appeared to have higher levels expression in gill tissues from Menez Gwen animals. The peptidoglycan recognition molecule encoding gene, PGRP, presented the highest level of transcriptional activity among the genes analyzed in Menez Gwen mussel gill tissues, seconded by carcinolectin and thus denoting the relevance of immune recognition molecules in early stage of the immune responses onset. Genes regarded as encoding molecules involved in signaling pathways were consistently expressed in both Menez Gwen and Lucky Strike mussel gill tissues. Remarkably, the immunity-related GTPase encoding gene demonstrated, in Lucky Strike samples, the highest level of expression among the signaling molecule encoding genes tested when expressions levels were compared between Menez Gwen and Lucky Strike animals. A differential expression analysis of bacterial genes between Menez Gwen and Lucky Strike mussels indicated a clear expression signature in the latter animal gill tissues. The bacterial community structure ensued from the 16S rRNA sequencing analyses pointed at an unpredicted conservation of endosymbiont bacterial loads between Menez Gwen and Lucky Strike samples. Taken together, our results support the hypothesis that B. azoricus exhibits different transcriptional statuses while living in distinct hydrothermal vent sites may result in distinct gene expressions because of physico-chemical and/or symbiont densities differences.
authors.affiliation: IMAR-Center, University of the Azores, 9901-862 Horta, Portugal; Department of Oceanography and Fisheries, University of the Azores, 9901-862 Horta, Portugal. Electronic address: raul@uac.pt.
authors.forename: Raul
authors.initials: R
authors.lastname: Bettencourt
authors.affiliation: IMAR-Center, University of the Azores, 9901-862 Horta, Portugal.
authors.forename: Mónica
authors.initials: M
authors.lastname: Rodrigues
authors.affiliation: IMAR-Center, University of the Azores, 9901-862 Horta, Portugal; Department of Oceanography and Fisheries, University of the Azores, 9901-862 Horta, Portugal.
authors.forename: Inês
authors.initials: I
authors.lastname: Barros
authors.affiliation: IMAR-Center, University of the Azores, 9901-862 Horta, Portugal; Department of Oceanography and Fisheries, University of the Azores, 9901-862 Horta, Portugal.
authors.forename: Teresa
authors.initials: T
authors.lastname: Cerqueira
authors.affiliation: Department of Oceanography and Fisheries, University of the Azores, 9901-862 Horta, Portugal.
authors.forename: Cátia
authors.initials: C
authors.lastname: Freitas
authors.affiliation: Department of Oceanography and Fisheries, University of the Azores, 9901-862 Horta, Portugal.
authors.forename: Valentina
authors.initials: V
authors.lastname: Costa
authors.affiliation: Biocant, Biotechnology Innovation Center, Cantanhede, Portugal.
authors.forename: Miguel
authors.initials: M
authors.lastname: Pinheiro
authors.affiliation: Biocant, Biotechnology Innovation Center, Cantanhede, Portugal.
authors.forename: Conceição
authors.initials: C
authors.lastname: Egas
authors.affiliation: IMAR-Center, University of the Azores, 9901-862 Horta, Portugal; Department of Oceanography and Fisheries, University of the Azores, 9901-862 Horta, Portugal.
authors.forename: Ricardo Serrão
authors.initials: RS
authors.lastname: Santos
mesh_terms: D000818:Animals; D001267:Atlantic Ocean; D001483:Base Sequence; D049872:Bivalvia / Q000378:metabolism* / Q000382:microbiology*; D019295:Computational Biology; D017931:DNA Primers / Q000235:genetics; D017753:Ecosystem; D005786:Gene Expression Regulation / Q000502:physiology*; D005843:Geography; D005880:Gills / Q000382:microbiology; D060086:Hydrothermal Vents; D017404:In Situ Hybridization, Fluorescence; D064307:Microbiota / Q000235:genetics*; D008969:Molecular Sequence Data; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D013559:Symbiosis; D059467:Transcriptome
chemical_list: D017931:DNA Primers; D012336:RNA, Ribosomal, 16S
keywords: Bathymodiolus azoricus; Endosymbiont; FISH; Gene expression; Immune genes