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    Bettencourt, R.; Rodrigues, M. I.; Barros, I.; Cerqueira, T.; Freitas, C.; Costa, V.; Pinheiro, M.; Egas, C.; Santos, R. S.;
    Project: FCT | PEst-OE/EEI/LA0009/2011 (PEst-OE/EEI/LA0009/2011), EC | HERMIONE (226354), FCT | PTDC/MAR/65991/2006 (PTDC/MAR/65991/2006)

    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 vent sites and in close vicinity off the Azores region near the Mid-Atlantic Ridge (MAR). The distinct relationships that vent mussels have developed with their physical and chemical environments are likely reflected in global gene expression profiles providing thus a means to distinguish geographically distinct vent mussels on the basis of gene expression studies, fluorescence in situ hybridization (FISH) experiments and 16S rRNA amplicon sequencing, to assess the natural expression of bacterial genes and vent mussel immune genes and the constitutive distribution and relative abundance of endosymbiotic bacteria within gill tissues. Our results confirmed the presence of methanotroph-related 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 sulfur-oxidizing-related nucleic-acid probe. Quantitative PCR (qPCR) studies revealed varied 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 MG or LS animals whereas the genes encoding effector molecules appeared to have higher levels expression in MG gill tissues. The peptidoglycan recognition molecule, encoding gene, PGRP presented the highest level of transcriptional activity among the genes analyzed in MG 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 MG and LS gill tissues. Remarkably, the immunity-related GTPase encoding gene demonstrated in LS samples, the highest level of expression among the signaling molecule encoding genes tested when expressions levels were compared between MG and LG animals. A differential expression analysis of bacterial genes between MG and LS indicated a clear expression signature in LS gill tissues. The bacterial community structure ensued from the 16S rRNA sequencing analyses pointed at a unpredicted conservation of endosymbiont bacterial loads between MG and LS samples. Taken together, our results support the premise that Bathymodiolus azoricus exhibits different transcriptional statuses depending on which hydrothermal vent site it is collected from and within the same collection site while exhibiting differential levels of expression of genes corresponding to different immune functional categories. The present study represents a first attempt to characterize gene expression signatures in hydrothermal vent animals issued from distinct deep-sea environmental sites based on immune and bacterial genes expressions.

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