Çédille, revista de estudios franceses

Presentació

Culture primaire de cellules de branchies de truite : caractérisation morphologique en microscopies photonique et électronique à transmission et à balayage

National audienc

Trout gill cells in primary culture on solid and permeable support

International audienc

Biological functions of trout pavement-like gill cells in primary culture on solid support : pHi regulation, cell volume regulation and xenobiotic biotransformation

International audienceThis review presents results obtained on rainbow trout gill cells in primary culture on solid support. Ultrastructural analysis showed that cultured gill cells displayed features of pavement cells in situ. Several biological functions have been investigated on these cultured cells. First, it was shown that their intracellular pH at rest and after acidosis is regulated by a Na+/H+ exchanger. Second, gill cells in primary culture can regulate their volume after a cell swelling. Intracellular calcium appears to be involved in this regulation. The effects of different xenobiotics on the capacity of gill cells to regulate their volume are presented. Third, cultured pavement cells contain biotransformation enzymes to metabolize xenobiotics. All these results demonstrate that gill cells in primary culture on solid support represent a promising in vitro model for the study of pavement cells physiology, In conclusion, applications of this culture are discussed and compared with the permeable filter method, together with the limitations and prospects of this in vitro model on solid support

Comparison of robustness traits in two isogenic lines of rainbow trout : use of a new approach based on modelisation of biological responses to acute stressors

Symposium : Stress in fishabsen

pHi regulation and ultrastructural analysis in cultured gill cells from freshwater or seawater-adapted trout

International audiencePrimary cultures of gill cells from freshwater and seawater-adapted trout were compared. These cultures, developed from an explant technique, exhibited a similar growth. Ultrastructural comparison between cultured and in situ cells showed that most of the cells in primary culture resembled the so called 'pavement' cells, whereas chloride cells were not observed in the cultured epithelium. Several other cells types, representing a minority of cells in primary culture, were observed (mucous cells, vesicular cells, cells with large dense granules and cells containing lysosomes). Morphological observations of cultured pavement cells from freshwater and seawater trout gills were similar, although the density of cellular organelles in cells was less under freshwater conditions. In addition to the morphological comparison, the regulation of intracellular pH in cultured cells from freshwater and seawater gills was examined. Resting pHi was not different for freshwater or seawater gill cells. A sodium-dependent and amiloride-sensitive mechanism was found in cultured cells. Under the experimental conditions used here, this mechanism was most likely a Na+/H+ antiporter in pavement cells from freshwater and seawater-adapted trout. The comparison of pHi recovery after acidification of cells from freshwater and seawater gills showed that the activity or the number of antiporters was higher for cells from seawater trout gill

A multivariate analysis using physiology and behavior to characterize robustness in two isogenic lines of rainbow trout exposed to a confinement stress

Robustness is a complex trait difficult to characterize and phenotype. In the present study, two features of robustness in rainbow trout were investigated: sensitivity and resilience to an acute stressor. For that purpose, oxygen consumption, cortisol release, group dispersion and group activity of two isogenic lines of juvenile rainbow trout were followed before and after an environmental challenge. The effect of a 4h confinement protocol (~140kg/m3), which is generally considered as a highly stressful challenge, was investigated. Temporal patterns produced by this experiment were analyzed using multivariate statistics on curve characteristics to describe physiological and behavioral adaptive systems for each isogenic line. The two isogenic lines were found to be highly divergent in their corticosteroid reactivity. However, no correlation between physiological and behavioral sensitivity or resilience was observed. Furthermore, the multivariate analysis results indicated two separate and independent fish group coping strategies, i.e. by favoring either behavioral or physiological responses. In addition, considerable intra-line variabilities were observed, suggesting the importance of micro-environment effects on perturbation sensitivities. In this context, cortisol release rate variability was found to be related to the pre-stress social environment, with a strong correlation between pre-stress aggressiveness and cortisol release rate amplitude. Overall, this approach allowed us to extract important characteristics from dynamic data in physiology and behavior to describe components of robustness in two isogenic lines of rainbow trout

Transcriptomic response of the hydrothermal mussel Bathymodiolus azoricus in experimental exposure to heavy metals is modulated by the Pgm genotype and symbiont content

International audienceHydrothermal vent mussels belonging to the genus Bathymodiolus dominate communities at hydrothermal sites of the Mid-Atlantic Ridge. The mussel Bathymodiolus azoricus harbors thiotrophic and methanotrophic symbiotic bacteria in its gills and evolves in naturally highly metal contaminated environments. In the context of investigations on metal tolerance/effect in B. azoricus, we focused our work on the short-term adaptive response (15 days) of mussels to different metals exposure at a molecular level using metal concentrations chosen to mimic natural situations at three vents sites. The expression of a set of 38 genes involved in different steps of the metal uptake, detoxication and various metabolisms was analysed by qPCR. Mussels were also genotyped at 10 enzyme loci to explore the relationships among natural genetic variation and gene expression. Relation between symbiont content (both sulfur-oxidizing and methanogen bacteria) and gene expression was also analysed. Our study demonstrated the influence of metal cocktail composition and time exposure on the transcriptome regulation with a specific pattern of regulation observed for the three metal cocktail tested. We also evidenced the significant influence of some specific Pgm genotype on the global gene expression in our experimental populations and a general trend of a higher gene expression in individuals carrying a high symbiont content. (C) 2014 Elsevier B.V. All rights reserved