The aquatic environment is severely impacted by xenobiotics that are released due to anthropogenic activities, threatening ecosystem health. Some of these contaminants accumulate in lipophilic fish tissues and are maternally transferred to developing offspring, affecting their growth and performance. However, knowledge about the long-term and generational impacts associated with maternal transfer of contaminants is limited in fish. In this thesis, the hypothesis tested was that maternal transfer of bisphenol A (BPA) leads to disruption in the developmental programing of growth and stress axes functioning in rainbow trout (Oncorhynchus mykiss), and that these changes are passed on to the next generation. This was tested by exposing oocytes to either control (vehicle; <0.01% ethanol) 0.3, 3.0, and 30.0 mg l-1 BPA in ovarian fluid for 3 h, prior to fertilization, to mimic maternal transfer. This led to the accumulation of 0, 0.8, 4.4 and 41.3 ng BPA embryo-1. Oocytes were fertilized with milt from clean males, and offspring growth, development and stress performances were assessed in a clean environment for a year (F1 generation). For F2 generation, oocytes collected from F1 females, raised from the different BPA accumulated eggs, were fertilized with milt from clean males and raised in a clean environment for one year as described for F1 generation. The accumulated BPA in eggs was quickly cleared and it was no longer detected in the F1 embryos at hatch. BPA exposure reduced specific growth rate and increased food conversion ratio in larvae reared from BPA-laden oocytes. Moreover, BPA-exposed fish had an altered cortisol developmental profile and a delay in stress axis maturation. In addition, the mRNA abundance of genes involved in somatotropic [insulin-like growth factor (IGF) -1; IGF-2; IGF receptor b (IGF-1rb)] and stress axes functioning [steroidogenic acute regulatory protein (StAR); cytochrome P450 side chain cleavage (P450scc)] were altered. Also, changes in thyroid signaling [thyroid receptor (TR) mRNA levels] and cortisol signaling [glucocorticoid receptor (GR) protein expression] were disrupted temporally during development. These results demonstrate that BPA accumulation in eggs, mimicking maternal transfer, impacts growth and development, and delays stress axis maturation via non-reproductive endocrine disrupting routes in trout. Some of the BPA changes seen in F1 generation also persisted in the F2 generation. For instance, ancestral exposure to BPA led to reduced growth and whole body glycogen content prior to feeding in the F2 fish. The developmental transcript profile of growth hormone-1and -2, IGF-1 and -2 and IGF-1rb, along with whole body cortisol levels were impacted by ancestral exposure to BPA. Moreover, a delay in cortisol dynamics post-stress was noted in the F2 fish of BPA exposure lineage. Our results show that ancestral exposure to BPA leads to effects on growth and stress performance in rainbow trout, but the mechanism is not known. To further investigate the long-term effect of BPA accumulation in eggs on stress performances, F1 and F2 juvenile fish were subjected to an acute stressor. Also, head kidney tissues from these juvenile fish were subjected to adrenocorticotrophic hormone (ACTH) stimulation in vitro to assess cortisol production capacity. BPA accumulation in eggs led to a reduced acute handling stressor-induced plasma cortisol response in trout from the F1 and F2 (only high BPA group) generations. Also, BPA exposure had a pronounced impact on acute handling stressor-mediated plasma glucose (only F2 generation) and lactate levels, indicative of a metabolic disturbance. BPA exposure (only the 4.4 ng group) did affect unstimulated but not stimulated [ACTH or 8-bromo-cyclic AMP (8-B-cAMP)] cortisol production from head kidney slices of juvenile fish from F1 generation. In the F2 generation, there was an increase in ACTH-stimulated cortisol production only from the high BPA-exposed group. Overall, BPA in eggs disrupts long-term cortisol and metabolic stress performances in rainbow trout. While the impaired plasma cortisol stress performance was dose-related in the F1, the effect was apparent only for high BPA group in the F2 generation, suggesting that the generational effects on cortisol stress axis functioning may be concentration-dependent. A metabolomics approach further confirmed multigenerational effects associated with BPA accumulation in eggs. Analysis of the metabolome profile at hatch and prior to first feed, using gas chromatography-time of flight-mass spectrometry (GC-TOF-MS), revealed a BPA-mediated metabolic disruption, including changes in pathways involved in carbohydrate, lipid and amino sugar metabolism, and amino acid metabolism and synthesis. Pathways involved in citric acid cycle and alanine, aspartate and glutamate metabolism were altered in both generations, suggesting that these pathways have the potential to be markers with predictive value for multigenerational effects of BPA in fish. Altogether, the study provides novel insights on the impact of BPA on rainbow trout metabolome at hatch and first feed. The results suggest that pathways involved in energy metabolism are targets for BPA impact and should be investigated as potential markers for BPA toxicity. Overall, BPA accumulation in oocytes induces long-term delays in growth and stress axis maturation in F1 generations fish, and these effects persist in the F2 generation. The developmental profiles of key genes of the somatotropic and HPI axes were altered by BPA, along with whole body composition, suggesting that BPA exposure leads to a metabolic disturbance in fish, resulting in reduced growth. Additionally, the altered plasma cortisol response to acute stress in F1 and F2 juveniles provides evidence for multigenerational effects of BPA on stress axis functioning. The current study proposes that BPA-induced epigenetic modifications during early development may be playing a key role in the generational effects on growth and stress axes disruption in trout. The finding that the growth and developmental changes to BPA exposure also corresponds with endocrine and metabolome changes in multiple generations in trout is novel, and underscores the necessity to develop new risk assessments tools for chemicals that are maternally transferred in fish.

L'évaluation de l'impact écologique d'une exposition aux rayonnements ionisants est devenue une préoccupation majeure. L'objectif de ce doctorat était d'étudier les effets multigénérationnels d'une irradiation gamma chronique selon une approche intégrée, des traits d'histoire de vie aux mécanismes subcellulaires chez un organisme modèle, le nématode Caenorhabditis elegans. L'étude des effets d'une irradiation gamma chronique sur les traits d'histoire de vie de C. elegans a d'abord été effectuée. Pour cela, trois générations ont été exposées et deux générations ont été placées en environnement « contrôle » après exposition parentale. Puis, différents mécanismes subcellulaires pouvant expliquer les effets observés sur les traits d'histoire de vie ont été par la suite caractérisés. Les résultats obtenus ont mis en évidence que (i) la reproduction était le critère d'effet le plus sensible, (ii) une augmentation de la radiosensibilité était observée sur trois générations exposées et (iii) les effets de la génération parentale étaient transmis aux générations non-exposées. Une augmentation de l'apoptose, une diminution du stock de spermatozoïdes et du nombre de cellules mitotiques semblent expliquer la diminution de la reproduction dans les générations exposées. Seule une diminution du nombre de spermatozoïdes a été observée en parallèle d'une diminution de la reproduction dans les générations placées non exposées. Ce projet de recherche a permis d'apporter des connaissances sur les effets multigénérationnels d'une irradiation gamma et montre l'intérêt d'utiliser une approche intégrée pour mieux comprendre les mécanismes d'action liés à l'action d'un polluant. / The environmental risk assessment of chronic exposure to ionizing has become a major concern. The aim of this PhD was to study the multigenerational effects of chronic gamma radiation in an integrated manner (to the life history traits from the subcellular mechanisms) in a model organism, the nematode Caenorhabditis elegans. First, studying the effects of chronic gamma radiation on the life history traits of C. elegans was performed. For that, three generations have been exposed to different dose rates and two generations have been placed in "control" environment after parental exposure. The second part of this thesis aimed to characterize the different subcellular mechanisms that could explain the observed effects on the life history traits after multigenerational exposure. The results showed that (i) the reproduction was the most sensitive endpoint to gamma radiation, (ii) an increase in radiosensitivity was observed over three exposed generations and (iii) the effects of the parental generation were transmitted to the non-exposed generations. An increase in apoptosis, a reduction in the stock of sperm, and to a lesser extent, a decrease in the number of mitotic cells, could explain the observed decrease in reproduction for the exposed generations. Only a decrease in sperm number was observed in parallel with a reduction in the cumulative number of larvae in the non-exposed generations. This research contributes to our knowledge on the multigenerational effects of gamma irradiation and shows the importance of an integrated approach to better understand the mechanisms of action related to the action of a pollutant and improve the environmental risk assessment.

Dans ce travail de thèse, nous avons évalué les effets neurotoxiques multigénérationnels de l’exposition des mères F0 gestantes et allaitantes aux polychlorobiphényles non-dioxin-like indicateurs (PCB-NDLi), à un profil mimant l'exposition humaine à partir de poissons contaminés, sur le développement et le comportement, y compris les réponses émotionnelles et les interactions sociales, des deux générations F1 et F2 des souris mâles et femelles, à différentes phases de leur ontogenèse. Deux faibles doses des PCB-NDLi : (i) la DJT, qui est de 10 ng/kg/j, et (ii) une dose environnementale de 1000 ng/kg/j, ont été administrés par accès libre aux souris mères F0. En fonction de la modalité d’exposition des parents F1 aux PCB, 4 groupes de génération F2 ont été obtenus, en croisant (i) des pères F1 exposés à des mères F1 non exposées, (ii) des mères F1 exposées à des pères F1 non exposés, (iii) des deux parents F1 exposés, ou (iv) des deux parents F1 non exposés (témoins), aux PCB en période périnatale. Nos résultats ont montré que les mâles adultes de la génération F1 ont manifesté un comportement dépressif-like ; alors que les mâles F2, issus uniquement des pères F1 exposés aux PCB, ont exhibé un comportement anti-dépressif-like, ce que suggère que l’exposition périnatale des souris F1 aux PCB-NDLi a induit une altération multigénérationnelle d’origine parentale du comportement de la résignation, et ce d’une façon sexe dépendante. De même, une altération sexe-dépendante de l’anxiété, a été détectée chez la génération F1 exposées durant la période périnatale aux PCB-NDLi comme uniquement les souris mâles d’âge moyen F1 ont développé un phénotype anxieux qui a été transmis aux souris mâles d’âge moyen F2, via leurs pères F1. En outre, une altération multigénérationnelle du comportement social a été détectée chez les souris mâles et femelles F1 et F2. D’une façon remarquable, chez la génération F2, des altérations comportementales dépendantes à la fois du sexe et de la dose, ont été trouvées, malgré l’absence d’effets chez leurs parents F1, effets qui dépendaient également de l’origine parentale, tels que la diminution significative du niveau de la préférence pour la nouveauté sociale chez les souris mâles F2, issues uniquement des mères F1 périnatallement exposées à la dose 10 ng/kg de PCB. Le dosage des biomarqueurs chez les souris d’âge moyen de la génération F1 a révélé une altération de nombreux paramètres biochimiques, y compris une augmentation du niveau de corticostérone et de l’activité de l’acétylcholinestérase / In this study, we evaluated the multigenerational neurotoxic effects of gestational and lactational exposure of F0 female mice to a representative mixture of the six indicator non-dioxin-like-polychlorinated biphenyls (NDL-PCBs) at environmentally low doses, a profile that closely mimics human exposure to contaminated fish. The tolerable day intake (TDI) of 10 ng/kg/day and a higher environmental dose of 1000 ng/kg/day were administered by free access to F0 mothers during pregnancy and lactation. Afterwards, the development and behavior, including emotional responses and social interactions, of the two F1 and F2 generations of Swiss male and female mice at different phases of their ontogenesis, were assessed. Depending on the mode of exposure of F1 parents to PCBs, four F2 generation groups were obtained by crossing (i) F1 fathers perinatally exposed with unexposed F1 mothers, (ii) F1 mothers perinatally exposed with unexposed F1 fathers, (iii) both F1 parents perinatally exposed, or (iv) both F1 parents perinatally unexposed (controls), to PCBs. Our results showed that F1 adult males showed depressive-like behavior whereas F2 adult males, coming from F1 mothers, perinatally exposed to PCBs, exhibited anti-depressive-like behavior. This result suggested an induction of a multigenerational alteration that was of parental origin, on the resignation behavior in a sex-dependent manner. Similarly, sex-selective anxious behavior was detected in F1 middle-aged males perinatally exposed to PCBs, which was transmissible to F2 middle-aged males, via their F1 fathers. Furthermore, a multigenerational alteration of social behavior was found in F1 and F2 male and female mice. Remarkably, some behavioral alterations in F2 generation were found, despite of the absence of effects in their F1 parents, such as a significant decrease in the level of preference for social novelty in F2 male mice, coming from F1 mothers perinataly exposed to 10 ng/kg of NDL-PCBs. The biomarker assays in F1 middle-aged mice revealed an alteration in many biochemical markers, including increased corticosterone levels and acetylcholinesterase activity in male as well as females.