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BMC Genomics. 2009; 10: 222.
Published online May 13, 2009. doi:  10.1186/1471-2164-10-222
PMCID: PMC2689276
Molecular identification of differentially regulated genes in the hydrothermal-vent species Bathymodiolus thermophilus and Paralvinella pandorae in response to temperature
Isabelle Boutet,1,2 Didier Jollivet,1,2 Bruce Shillito,3 Dario Moraga,4 and Arnaud Tanguycorresponding author1,2
1CNRS, UMR 7144, Adaptation et Diversité en Milieu Marin, Station Biologique, de Roscoff, 29682 Roscoff, France
2UPMC Univ Paris 06, UMR 7144, Equipe Génétique et Adaptation en Milieu Extrême, Station Biologique de Roscoff, 29682 Roscoff, France
3UPMC Université Paris 6, UMR 7138, Systématique, Adaptation et Evolution, 75005 Paris, France
4UMR CNRS 6539, Laboratoire des Sciences de l'Environnement Marin, Institut Universitaire Européen de la Mer, Université de Bretagne Occidentale, Place Nicolas Copernic, 29280 Plouzané, France
corresponding authorCorresponding author.
Isabelle Boutet: boutet/at/sb-roscoff.fr; Didier Jollivet: jollivet/at/sb-roscoff.fr; Bruce Shillito: Bruce.Shillito/at/snv-jussieu.fr; Dario Moraga: dario.moraga/at/univ-brest.fr; Arnaud Tanguy: atanguy/at/sb-roscoff.fr
Received July 24, 2008; Accepted May 13, 2009.
Abstract
Background
Hydrothermal vents and cold seeps represent oases of life in the deep-sea environment, but are also characterized by challenging physical and chemical conditions. The effect of temperature fluctuations on vent organisms in their habitat has not been well explored, in particular at a molecular level, most gene expression studies being conducted on coastal marine species. In order to better understand the response of hydrothermal organisms to different temperature regimes, differentially expressed genes (obtained by a subtractive suppression hybridization approach) were identified in the mussel Bathymodiolus thermophilus and the annelid Paralvinella pandorae irlandei to characterize the physiological processes involved when animals are subjected to long term exposure (2 days) at two contrasting temperatures (10° versus 20°C), while maintained at in situ pressures. To avoid a potential effect of pressure, the experimental animals were initially thermally acclimated for 24 hours in a pressurized vessel.
Results
For each species, we produced two subtractive cDNA libraries (forward and reverse) from sets of deep-sea mussels and annelids exposed together to a thermal challenge under pressure. RNA extracted from the gills, adductor muscle, mantle and foot tissue were used for B. thermophilus. For the annelid model, whole animals (small individuals) were used. For each of the four libraries, we sequenced 200 clones, resulting in 78 and 83 unique sequences in mussels and annelids (about 20% of the sequencing effort), respectively, with only half of them corresponding to known genes. Real-time PCR was used to validate differentially expressed genes identified in the corresponding libraries. Strong expression variations have been observed for some specific genes such as the intracellular hemoglobin, the nidogen protein, and Rab7 in P. pandorae, and the SPARC protein, cyclophilin, foot protein and adhesive plaque protein in B. thermophilus.
Conclusion
Our results indicate that mussels and worms are not responding in the same way to temperature variations. While the results obtained for the mussel B. thermophilus seem to indicate a metabolic depression (strong decrease in the level of mRNA expression of numerous genes) when temperature increased, the annelid P. pandorae mainly displayed a strong regulation of the mRNA encoding subunits and linkers of respiratory pigments and some proteins involved in membrane structure. In both cases, these regulations seem to be partly due to a possible cellular oxidative stress induced by the simulated thermal environment (10°C to 20°C). This work will serve as a starting point for studying the transcriptomic response of hydrothermal mussels and annelids in future experiments in response to thermal stress at various conditions of duration and temperature challenge.
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