This group of experiments focused on relationships between diet, somatotropic axis, and growth. Growth hormone (GH) and insulin-like growth factor I (IGF-I) are factors in the somatotropic axis, and important to development of growth cartilage in the young animal. The entire study was divided into four main experiments. Characteristics of growth in 113 Thoroughbred foals born over a five year period were described with a series of empirical and physiological equations. Glycemic and insulinemic responses to different feed compositions were evaluated with glycemic response tests. The 24 hr pattern of plasma glucose, insulin, GH, and IGF-I was described in yearlings fed two meals a day. Finally, an association between ADG and IGF-I was described in Thoroughbreds from birth to 16 mo of age. Feeding diets to the foal that influence the somatotropic axis during growth may affect development of growth cartilage in unexpected or detrimental ways. The pattern of weight in Thoroughbred foals from birth to 16 mo of age was most closely described by multiple regression with a combination of age, girth, body length, and physeal circumference (R2 = 0.99). Glycemic and insulinemic responses were significantly higher in yearlings fed a sugar and starch supplement when compared to those fed a fat and fiber supplement (P = 0.043 and 0.031; respectively). Glucose and insulin secretion was significantly affected by the feeding of two meals in a 24 hr period (P < 0.0001). Plasma IGF-I was positively correlated with ADG from birth to 16 mo of age in foals fed either a fat and fiber or sugar and starch supplement (r = 0.34, P < 0.0001). The results from the series of experiments described here suggest a possible role of dietary mangement in reducing the risk of skeletal disorders that involve the influence of IGF-I on chondrocyte maturation. / Ph. D.

Two trials were conducted to evaluate the effects of gender on optimal CP concentrations (TRIAL 1) and gender and anabolic implants on the somatotropic axis in feedlot cattle (TRIAL 2). More specifically, the objective of TRIAL 1 was to examine the effects of 3 dietary CP concentrations on performance, carcass characteristics, and serum urea nitrogen (SUN) in finishing steers and heifers and the objective of TRIAL 2 was to evaluate the effects of ovariectomy (OVX) and implantantation (200 mg of trenbolone acetate and 28 mg of estradiol benzoate; Synovex-Plus) on performance, serum urea nitrogen (SUN), serum IGF-1, and mRNA expression of hepatic IGF-1, GH receptor, and E receptor-α as well as pituitary GH, E receptor-α and GHRH receptor in feedlot heifers. TRIAL 1 results indicated that ADG was optimized when both steers and heifers were fed 12.5% CP and G:F was optimized for steers fed 12.5% CP but heifer G:F was optimal at 14.0% CP. Feeding diets containing 11.0% CP appears to cause a protein deficiency in both steers and heifers. TRIAL 2 data indicated that gender had no influence on performance or SUN in feedlot heifers. Serum IGF-1 is increased more in OVX heifers than intact heifers due to a greater response to implantation from the OVX heifers. However, the reason for the extra increase in serum IGF-1 is not clear, although trends in gene expression analysis suggest the possibility that the increased serum IGF-1 may be controlled outside of the somatotropic axis. Further research is warranted to examine the effects of OVX and anabolic implants on the somatotropic axis.

La nutrition au cours de la période postnatale précoce programme l’activité de l’axe somatotrope à l’âge adulte (GH/IGF-1). Une altération de la nutrition chez les souriceaux au cours de la lactation altère la croissance staturo-pondérale de façon permanente et augmente leurs susceptibilités à développer des pathologies cardio-métaboliques à l’âge adulte. La restriction au cours de la lactation induit une diminution des taux plasmatiques en IGF-1 et en leptine. Ceci est associé à une diminution transitoire de l’innervation de l’éminence médiane par les neurones GHRH, ce qui induit une hypoplasie hypophysaire permanente en cellules somatotropes. Durant ma thèse, j’ai étudié l’impact de la nutrition périnatale sur la mise en place de l’axe somatotrope, notamment les mécanismes impliqués dans la régulation du développement des neurones GHRH. Les cultures d’explants de noyaux arqués issus de souriceaux normalement nourris indiquent que l’IGF-1 stimule de façon préférentielle la croissance axonale des neurones GHRH par l'intermédiaire des voies PI3K/AKT et MAPK. La leptine présenterait quant à elle un effet plus global sur les neurones du noyau arqué, stimulant la croissance axonale des neurones GHRH et des neurones orexigène à NPY/AgRP. Les neurones GHRH issus de souris restreintes sont quant à eux résistants à la stimulation de la croissance axonale par l’IGF-1 ou la leptine. Concernant l’IGF-I, cette résistance est associée à une altération des capacités de phosphorylation de la voie PI3K/AKT, tandis que celles de l’IGF-1R et de la voie MAPK ne sont pas altérées. / Nutrition during lactation programs the activity of the somatotropic axis (GH/IGF-1). Alteration of nutrition during the early postnatal period in mice induces increased susceptibility to develop cardiovascular and metabolic pathologies later in life. Nutritional restriction during lactation permanently alters growth of mice. Ten days old restricted pups present decreased plasmatic level of IGF-1 and Leptin. They also present a transient alteration of median eminence innervation by GHRH neurons, which induce a permanent somatotroph cells (GH) hypoplasia in pituitary. The aim of my thesis was to study the impact of nutrition during the perinatal period on the programming of the somatotropic axis, notably the cellular and molecular mechanisms involved in the regulation of GHRH neuronal development. In vitro cultures of arcuate nucleus explants of hypothalamus from normally fed pups indicate that IGF-1 preferentially stimulates axonal growth of GHRH neurons by its signaling pathways PI3K/AKT and MAPK. Leptin present a more global effect and is able to stimulate axonal growth of arcuate nucleus neurons, including GHRH and AgRP neurons. GHRH neurons from restricted pups are resistant to the stimulation of axonal growth by IGF-1 or leptin. Regarding IGF-1, this resistance is associated with an alteration of phosphorylation capacities of the PI3K/AKT pathway, whereas those from IGF-1R and MAPK are not altered.

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.

Les processus physiopathologiques qui mènent à la dégénérescence neuronale ainsi qu’aux symptômes de la maladie de Huntington (MH) demeurent non identifiés et les hypothèses actuelles ne permettent pas d’expliquer l’hétérogénéité intra et interindividuelle de l’évolution de ces symptômes. Ainsi, la progression de la maladie reste donc difficile voire impossible à prédire. Dans ce contexte, il est important d’explorer d’autres facteurs qui semblent être impliqués dans le processus pathogène de la maladie mais qui pourraient également influencer l’évolution de ces symptômes et ainsi prédire la progression de la maladie. Plusieurs éléments de preuve renforcent l’hypothèse de l’existence de troubles hormonaux dans la MH tels que l’atteinte de l’hypothalamus et la perte de poids. Cependant, en raison du peu d’études, de leur qualité et de la discordance de leurs résultats, l’existence des modifications hormonales dans la maladie de Huntington et plus particulièrement leur lien avec la progression de la maladie reste controversée. L’objectif de ce travail est de décrire le profil hormonal de l’axe hypothalamohypophysaire dans la MH afin de mieux comprendre le rôle de ces hormones sur la progression et éventuellement sur la physiopathologie de la maladie. Dans notre étude transversale, nous avons mis en évidence une activation de l’axe somatotrope (Growth Hormone/Insulin Growth Factor 1), une inhibition en fonction de la sévérité de la maladie de deux axes : gonadotrope (Testostérone) et thyréotrope (Thyroid Stimulating Hormone et triiodothyronine) mais aucune modification des hormones de l’axe corticotrope ni de la prolactine. De plus, la modification hormonale de l’axe somatotrope était non pathologique et précoce alors qu’elle était tardive pour les deux autres axes. Pour expliquer le lien entre ces modifications et la progression de la maladie une étude longitudinale a été mise en place. Les résultats de cette étude montre que seule l’élévation plasmatique d’IGF1 était prédictive de la détérioration cognitive. L’ensemble de nos résultats apporte une meilleure description et compréhension du profil de l’axe hypothalamo-hypophysaire dans la maladie de Huntington. Les axes pituitaires ne sont pas tous atteints et leur atteinte n’est pas dans le même sens. La relation inverse entre l’activation de l’axe somatotrope et la détérioration cognitive renforce l’hypothèse d’une résistance à l’effet de l’IGF1 dans la maladie de Huntington comme pour la maladie d’alzheimer. En conclusion, compte tenu de l’implication de l’IGF1 dans la prédiction de la progression cognitive dans la maladie de Huntington, il serait intéressant de détecter si les modifications biologiques de l’IGF1 existent dès la phase asymptomatique cognitive afin d’envisager d’utiliser l’IGF1 comme biomarqueur de l’apparition ou de l’évolution des symptômes cognitives. D’un autre côté, il serait important d’étendre les recherches sur les mécanismes responsables des modifications hormonales dans la maladie de Huntington afin de mieux comprendre l’effet de cause à effet s’il existe entre ces modifications et les symptômes de la maladie / The pathophysiological processes leading to neurodegeneration and the symptoms of Huntington's disease (HD) remain unidentified and current hypothesis do not explain the intra and interindividual heterogeneity of the evolution of these symptoms. Thus, the progression of the disease remains difficult or impossible to predict. In this context, it is important to explore other factors that appear to be involved in the pathogenic process of the disease but could also influence the evolution of these symptoms and predict disease progression. Several evidences reinforce the hypothesis of the existence of hormonal disorders in HD such as the atrophy of the hypothalamus and weight loss. Because of few studies, their quality and the discrepancies of their results, the existence of hormonal changes in Huntington's disease and particularly their relationship to disease progression remains controversial. The objective of this work is to describe the hormonal profile of the hypothalamicpituitary axis in HD in order to better understand the role of these hormones on the progression and on the pathophysiology of the disease. In our cross-sectional study, we identified an activation of the somatotropic axis (Growth Hormone / Insulin Growth Factor 1), an inhibition according to the severity of the disease in two axes: gonadotrope (Testosterone) and thyréotrope (Thyroid Stimulating hormone and triiodothyronine) but no change in hormones of corticotropic axis and prolactin. In addition, the somatotropic axis is overactive even in patients with early disease. To explain the link between these changes and the progression of the disease, a longitudinal study was done. The results of this study showed that only the elevated plasma IGF1 was predictive of cognitive impairment. All of our results provide a better description and understanding of the profile of the hypothalamic-pituitary axis in Huntington's disease. Pituitary axes are not all disturbed. The inverse relationship between activation of the somatotropic axis and cognitive impairment strengthens the hypothesis of a resistance to the effect of IGF1 in Huntington's disease like in Alzheimer's disease. In conclusion, given the involvement of IGF1 in the prediction of cognitive progression in Huntington's disease, it would be interesting to detect whether the biological changes of IGF1 are already present at the asymptomatic cognitive stage in order to use IGF1 as a biomarker of the onset or changes in cognitive symptoms. On the other hand, , it would be important to extend research on the mechanisms responsible for hormonal changes in Huntington's disease to better understand the link between these changes and symptoms of the disease

Einleitung: Futterrationen für Hochleistungsmilchkühe basieren häufig auf Maissilage und liefern somit nur wenig frisches Gras, was eine niedrige Zufuhr an n-3-Fettsäuren, v.a. an α-Linolensäure (ALA), bewirkt. Dies führt einerseits zu einem reduzierten Status an ALA und konjugierter Linolsäure (CLA) und andererseits zu einem hohen n-6/n-3-Verhältnis bei laktierenden Kühen. Ziel der Untersuchungen: Das Ziel der vorliegenden Studie war es, die Einflüsse einer Supplementierung von essentiellen Fettsäuren (EFA, v.a. ALA) zusammen mit CLA auf den Fett- säurestatus, die Leistung, auf das antioxidative und inflammatorische System bei Milchkühen, die mit einer Mais-basierten Ration gefüttert wurden, in etablierter Laktation zu untersuchen und ver- schiede Stoffwechselwege, einschließlich der somatotropen Achse, näher zu beleuchten. Tiere, Material und Methoden: Es wurden 4 Kühe (3. Laktation, 126 ± 4 Tage in Milch) in einem 4 × 4 Latin Square untersucht. Die Kühe erhielten täglich abomasale Infusionen an Kokosöl (CTRL, 38,3 g/d; v.a. gesättigte Fettsäuren), Lein- und Distelöl (EFA, 39,1 und 1,6 g/d), Lutalin® (cis-9,trans-11 und trans-10,cis-12 CLA, jeweils 4,6 g/d) oder EFA+CLA für jeweils 6 Wochen. Die initiale Dosis wurde jeweils nach 2 Wochen verdoppelt, was in 3 Dosierungen resultierte (Dosis 1, 2 und 3). Es schloss sich eine 3-wöchige Washout-Periode an. Den Kühen wurde eine Mais-basierte Ration mit einem hohen n-6/n-3-V erhältnis (11,3:1) gefüttert. Die Trockensubstanzaufnahme und die Milchleistung wurden täglich und die Milchzusammensetzung wöchentlich gemessen. Die Fettsäuremuster im Milchfett und im Blutplasma, Plasmakonzentra- tionen von Metaboliten und Hormonen sowie von Parametern des antioxidativen Systems und der Immunantwort (nur in Woche 0 und 6) wurden jeweils in Behandlungswoche 0, 2, 4 und 6 analysiert. Lebergewebe wurde zu Beginn der Studie und jeweils nach 6 Wochen Behandlung entnommen und der Energiestoffwechsel sowie Parameter des antioxidativen Systems und der Immunantwort wurden auf Ebene der Transkription untersucht. Die statistische Auswertung wurde mittels ANOVA und der MIXED Prozedur (repeated measurements) in SAS durchgeführt, wobei Behandlung, Dosis und deren Interaktion als fixe Effekte und die Laktationswoche als Kovariable dienten. Ergebnisse: Die jeweils infundierten Fettsäuren stiegen sowohl im Plasma als auch in der Milch dosisabhängig an. Das n-6/n-3-Verhältnis des Milchfetts lag in der CTRL- und in der CLA- Gruppe höher als in den beiden EFA-Gruppen. Die Energie-korrigierte Milch und das Milchfett nahmen dosisabhängig in den beiden CLA-Gruppen ab. Es gab einen Trend für eine höhere Energiebilanz in der CLA-Gruppe. Der Milchproteingehalt war in den beiden CLA-Gruppen niedriger als in der CTRL-Gruppe und die Milchharnstoffkonzentration sank in beiden CLA- Gruppen dosisabhängig ab. Die Citratkonzentration in der Milch stieg dosisabhängig in der CLA- Gruppe an. Die Aktivität der Glutathionperoxidase im Blutplasma war in der CTRL-Gruppe ge- ringer als in der EFA-Gruppe und die Plasmakonzentration von β-Carotin stieg in beiden EFA- Gruppen mit der Dosis an. Die Plasmakonzentration des Gesamtcholesterols stieg dosisabhängig in allen Gruppen, außer der CLA-Gruppe, an. Die Plasmakonzentration des High-density- lipoprotein-Cholesterols stieg in der CTRL-Gruppe an und lag höher als in der EFA- und der CLA-Gruppe, während die Konzentrationen des Low-density-lipoprotein-Cholesterols dosisab- hängig in der EFA- und der EFA+CLA-Gruppe anstiegen und höher als in der CLA-Gruppe waren. Die hepatische Genexpression der 3-Hydroxy-3-methylglutaryl-CoA Synthase 1 wurde in allen Gruppen hochreguliert und lag in der EFA+CLA-Gruppe am höchsten. Die Genexpression des sterol regulatory element-binding factor 1 zeigte einen Trend für die niedrigsten Werte in den beiden EFA-Gruppen. Die Expression des leberspezifischen growth hormone receptor 1A (GHR1A) tendierte zu einer Erhöhung in der EFA+CLA-Gruppe im Vergleich zur CTRL-Gruppe. Die insulin-like growth factor I (IGF-I)-Plasmakonzentration stieg in der CLA-Gruppe an und der Plasmaspiegel des insulin-like growth factor binding protein 2 (IGFBP-2) lag in der EFA+CLA- Gruppe niedriger als in der CTRL-Gruppe. Die Albumin- und Harnstoffkonzentrationen im Plasma waren in der CLA-Gruppe niedriger als in der CTRL-Gruppe. Schlussfolgerungen: Die Milchfettdepression und das erhöhte Milch-Citrat weisen auf eine redu- zierte de-novo-Fettsäuresynthese und einen verbesserten oxidativen Status in der Milch durch CLA-Supplementierung hin. Weder CLA- noch EFA-Gaben zeigten eindeutige Wirkungen auf den Entzündungsstatus bei den Milchkühen. Die Supplementierung von EFA und CLA hatte Ein- fluss auf den Cholesterol- und den Fettstoffwechsel sowie deren Regulierung. Der erhöhte IGF- I-Plasma-Spiegel in der CLA-Gruppe sowie die niedrigere IGFBP-2-Plasmakonzentration und die erhöhte Genexpression des GHR1A in der Leber der EFA+CLA-Gruppe deuten auf stimulierende Effekte auf die somatotrope Achse hin. Weiterhin scheinen CLA-Gaben auch den Proteinstoffwechsel von Milchkühen zu beeinflussen.