O aumento da obesidade infantil tem sido correlacionado com a prevalência da puberdade precoce. Em animais pré-puberes verifica-se que a administração da dieta hiperlipídica induz a antecipação da puberdade. Estudos sugerem que a leptina pode influenciar o eixo reprodutivo por meio de sua ação em neurônios que expressam kisspeptina, no hipotálamo. Uma subpopulação de neurônios kisspeptidérgicos do núcleo arqueado coexpressa a dinorfina e neuroquinina B, que exercem uma ação autorreguladora sobre a secreção de kisspeptina. Considerando que a leptina, a kisspeptina, neuroquinina B e dinorfina são apontados como reguladores do início da puberdade e da função reprodutiva, este trabalho investigou como a dieta hiperlípidica (HFD) pode influenciar esses moduladores ao longo do início do desenvolvimento puberal e após a maturação sexual. Camundongos fêmeas Kiss1-hrGFP foram tratadas com a HFD ou dieta controle a partir do desmame e foram decapitadas aos 28, 32 e 63 dias (Diestro II) e o sangue coletado para dosagem plasmática de leptina, LH e estradiol. Um segundo grupo de fêmeas, tratadas com HFD ou dieta controle, receberam com 32 e 63 dias (Diestro I) uma injeção intraperitoneal de leptina (0,5 mg/kg) ou de veículo, 45 minutos após a injeção foram anestesiadas e submetidas à perfusão cerebral. Cortes hipotalâmicos foram utilizados para realização da técnica de imunoistoquímica para dupla marcação: Kiss1-p-STAT3/ p-STAT3-CART/ Kiss1-Dyn e Kiss1-NKB. Verificamos que a HFD administrada após o desmame, quando comparada à dieta controle, antecipou a abertura vaginal, o primeiro estro e o ínicio da ciclicidade em fêmeas Kiss1-hrGFP. Como indicativo de ativação do eixo reprodutivo neste grupo, foi observado aumento do peso do útero e ovário, aos 32 dias. Esses efeitos estiveram associados com: a) o aumento da concentração plasmática de leptina; b) aumento da ativação de p-STAT3 no núcleo arqueado (ARC) e c) aumento de p-STAT3 em neurônios Kiss1-hrGFP e em neurônios CART do ARC em resposta ao estímulo da leptina. A HFD não afetou a colocalização entre Kiss1-Dyn e Kiss1-NKB em fêmeas no início do desenvolvimento puberal. Em fêmeas Kiss1-hrGFP (63 dias) a administração da HFD por 6 semanas não alterou a regularidade do ciclo estral, houve elevação da leptina plasmática associada com diminuição da expressão da p-STAT3 no ARC em resposta ao estímulo com leptina, sem alterar a responsividade dos neurônios Kiss1-hrGFP a este hormônio. Em conclusão, 9 os resultados demonstram que a antecipação da puberdade induzida pela HFD está associada com o aumento da ação central da leptina, que ao estimular o sistema kisspeptidérgico, direta ou indiretamente, por meio de neurônios CART, pode aumentar o tônus excitatório sobre o eixo reprodutivo. A administração da HFD após o desmame (6 semanas) promoveu uma redução à resposta da leptina no ARC na vida adulta, porém não afetou a responsividade dos neurônios Kiss1-hrGFP a este hormônio e nem a regularidade do ciclo estral nas fêmeas Kiss1-hrGFP. Em conjunto, esses resultados indicam que as ações da leptina nesses neurônios foram preservadas, mesmo com a resistência parcial de suas ações no ARC. Estudos adicionais deverão ser realizados para se avaliar os efeitos a longo prazo da HFD sobre o eixo reprodutivo. / The increase of childhood obesity has been correlated with the early onset of puberty. In prepubertal animals it appears high fat diet (HFD) induces anticipation of puberty. Several studies have indicated that the effect of leptin on reproductive axis can be mediated by its action on Kiss1 neurons in the hypothalamus.A subpopulation of Kiss1 neurons in the arcuate nucleus also shows co-expression of neurokinin B and dynorphin A,presenting a self-regulatory action kisspeptin secretion. Since leptin, kisspeptin, neurokinin B and dynorphin are identified as regulators of puberty onset and reproductive function, this study investigated the influence of high fat diet on these modulators along puberty onset and after sexual maturation. A group of Kiss1-hrGFP-female mice received the HFD or regular chow after weaning. The Kiss1-hrGFP females were decapitated at 28, 32 and 63 days old (Diestrus II) and blood was collected for measurement of plasma leptin, LH and estradiol. A second group of females, treated with HFD or chow after weaning, received at 32 and 63 days old (Diestrus I) an intraperitoneal injection of leptin (0.5 mg / kg) or vehicle, and 45 minutes after they were anesthetized and subjected to cerebral perfusion. Immunohistochemistry was performed for double labeling: Kiss1-p-STAT3/ p-STAT3-CART/ Kiss1-Dyn and Kiss1-NKB in the hypothalamus. It was found that, compared to controls; the HFD anticipated the vaginal opening, the first estrus and the beginning of cyclicity in Kiss1-hrGFP females. An increase in the uterine weight and ovaries, confirmed the activation of reproductive axis in the HFD group at 32 days old. These effects were associated with: a) an increase in plasma leptin concentration; b) the activation of p-STAT3 in the arcuate nucleus (ARC) and c) the increase of p-STAT3 in Kiss1-hrGFP neurons and in CART neurons of the ARC in response to the stimulus of leptin. HFD didnt affect colocalization between Kiss1-Dyn and Kiss1-NKB in females mice at hte pubertal onset . Kiss1-hrGFP female mice (63 days old), exposed to HFD for 6 weeks,exhibited regular estrous cycles, increased plasma levels of leptin associated with a decreased p-STAT3 expression in the ARC after leptin stimulation, with no changes in the responsiveness of Kiss1-hrGFP neurons to this hormone, compared to controls. In conclusion, 11 the results demonstrate that the early puberty induced by the HFD is associated with increased central leptin action, by stimulating the kisspeptidergic system, directly or indirectly, through CART neurons that can increase the excitatory tone in the reproductive axis. The administration of HFD after weaning (6 weeks) induced a decrease on the response to leptin in the ARC in adulthood, but without changes in the responsiveness of Kiss1-hrGFP neurons to this hormone and the regularity of the estrous cycle in Kiss1-hrGFP females. Taken together, these results indicate that the actions of leptin in kisspeptin neurons were preserved after 6 weeks of HFD, despite the partial resistance to this hormone in the ARC. Additional studies should be conducted to evaluate the long-term effects of HFD in the reproductive axis.

Aquaculture is an essential developing sector for world food production however one of the major bottlenecks for the sustainability of the aquaculture industry is the ability to control fish reproduction in captivity and to produce high quality seeds. European sea bass is a one of most commercially important species for the European fish farming industry. If broodstock management under captivity is well established, problems remain in hatcheries where survival can be low and deformity prevalence high as well as in on growing sites where fish reach puberty early especially with skewed sex ratio towards males. Sea bass displays strong seasonality in its physiology and is therefore an excellent candidate for the study of the photo-neuroendocrine control of reproduction and growth. The overall aims of this thesis were to better understand the molecular and endocrine drivers that control the Brain-Pituitary-Gonad axis in repeat spawner sea bass, and expand our knowledge of sea bass light and temperature regulation of melatonin production. First, this PhD project investigated the seasonal expression of kisspeptin, GnRH and gonadotropin genes in relation to the gonadal development throughout a reproductive cycle in male repeat spawning sea bass (Chapter 3). A partial sequence for the receptor kissr4 was isolated and described showing similarity to all other teleost species sequences available to date. QPCR molecular assays were validated to mesure the expression of a suite of genes along the BPG axis including kisspeptin related genes (Kiss1 and Kiss2 and its receptor kissr4) over a full reproductive cycle (12 months) in adult male European sea bass. Brain Kisspeptin mRNA expression levels (kiss1, 2 and kissr4) showed clear seasonal profiles and correlated well to other BPG markers (GnRHs, fshβ and lhβ), supporting a possible involvement of kisspeptin genes in the seasonal control of reproduction in repeat spawning sea bass. Moreover, clear seasonal patterns were observed for expression of the genes encoding for pituitary mRNA expression of lhβ and fshβ, with a significant correlation between expression of both subunits and GSI and steroids levels. However, no clear seasonal profiles in brain GnRHs gene expression were observed with the exception to some peaks in GnRH1 and GnRH2. The second part of this PhD project investigated the potential direct effect of the two kisspeptin core peptides (kiss1 and kiss2) on the pituitary gonadotropin gene expression (Chapter 4). The aim of this work was to better understand the mechanism by which kisspeptin acts on the BPG axis. This was done by testing the kisspeptin decapeptide core sequences on the lhβ and fshβ transcript expression in primary culture of sea bass pituitary cells using QPCR technique. The findings, as a whole, provided evidence that kisspeptin can act directly on the pituitary gonadotroph cells and modulate fshß and lhß mRNA expression in sea bass although effects were limited and not uniform. Of note, kissr4 gene expression was also detected in the sea bass pituitary. The third part of this PhD project focused on the effects of environmental signals (photoperiod and temperature) on melatonin production (Chapter 5). Environmental manipulation is routinely used in the aquaculture industry with the purpose of enhancing growth and manipulating the timing of reproduction in seasonal fish species like sea bass. Melatonin, known as the light perception and time keeping hormone, has been suggested to play key roles in the synchronisation of most physiological functions in vertebrates, although the mechanisms by which melatonin controls reproduction, growth and behaviour are still not fully understood in fish. The studies performed aimed .to determine the synergistic effects of both temperature and photoperiod on the daily phase and amplitudinal changes in melatonin production through both in vivo and in vitro trials. The results confirmed the diel melatonin rhythm in sea bass as previously reported in many teleost species with “high at night” and “low at day” melatonin profiles. Temperature showed clear effects on the amplitude of the melatonin production under both in vivo and in vitro conditions for both long day and short day photoperiods. Furthermore, no endogenous melatonin production was found under constant darkness in both in vivo and in vitro conditions. These results suggested a lack of intrapineal (or located elsewhere such as retina and/or deep brain) oscillators in sea bass, contrasting with previous reports. These results further enhance our knowledge of light perception and circadian rhythmicity in sea bass, while the circadian system remains to be characterised in sea bass and teleosts as a whole. Overall, this doctoral work broadened our understanding on the photoneuroendocrine control of reproduction in a seasonal fish species, sea bass. New knowledge gained and tools developed from this work should help to develop/optimise husbandry techniques for the sea bass farming industry with the view to increase production and profitability and thus promoting the sustainable expansion of the sea bass aquaculture in Europe. It has also the potential to help the fishery sector in the modelling of wild sea bass populations.

Female reproductive senescence is widespread among mammalian species, but menopause is limited to species with menstrual cycles. While hormonal changes at menopause have profound impacts in the lives of women at middle age, the complex mechanisms underlying this process remain obscure. All three levels of the hypothalamic-pituitary-gonadal (HPG) axis are involved in reproductive aging, and evidence highlights a critical role for the dysregulation of gonadotropin-releasing hormone (GnRH) neurons, the hypothalamic cells that drive reproductive function. To investigate neuroendocrine mechanisms that may initiate and perpetuate reproductive decline at each step in the transition to acyclicity, I utilized an ovarian-intact middle-aged female rat model of natural reproductive senescence. These studies focused on three hypothalamic nuclei that are known to control GnRH activity: the anteroventral periventricular nucleus (AVPV), the site of positive hormone feedback onto GnRH neurons; the arcuate nucleus (ARC), the site of negative feedback; and the median eminence (ME), the site of GnRH release, with the following specific aims: 1) Characterize neuroendocrine gene and protein expression in female rats throughout the natural transition to acyclicity; 2) Determine the effects of chronic N-methyl-D-asparate receptor subunit 2b (NMDAR-NR2b) inhibition in acyclic females; and 3) Examine neuroendocrine gene expression during premature reproductive senescence after perturbation of the HPG axis. The results of these studies identified novel molecular and cellular changes with age and reproductive cycle status in the ARC and ME, two regions that are underappreciated for their roles in reproductive senescence. Surprisingly, few molecular targets were identified in the AVPV, a region that is much better-studied in this context. In the ME and ARC, I found changes in transcription factors and evidence of altered hormone feedback via changes in sex steroid hormone receptors and enzyme expression with reproductive aging. I also discovered decreased expression of genes for the excitatory neuropeptides, kisspeptin and neurokinin B, as well as decreased percentage of kisspeptin immunoreactive cells and their co-expression with estrogen receptor alpha in the ARC. And finally, in the ME, neurotrophic factor expression was changed with age, and the presence and phosphorylation state of the NR2b subunit of the NMDA receptor contributes to a greater inhibitory tone with acyclicity. Together these studies have identified novel pathways, especially in the ARC and ME, that are related to reproductive decline. Furthermore, changes in the hypothalamic neural and glial network of neurotransmitters, neuropeptides, hormone receptors and other transcription factors are likely contributing to altered responses to hormonal feedback and decreased excitatory drive for GnRH release. / text

No description available.

O aumento da obesidade infantil tem sido correlacionado com a prevalência da puberdade precoce. Em animais pré-puberes verifica-se que a administração da dieta hiperlipídica induz a antecipação da puberdade. Estudos sugerem que a leptina pode influenciar o eixo reprodutivo por meio de sua ação em neurônios que expressam kisspeptina, no hipotálamo. Uma subpopulação de neurônios kisspeptidérgicos do núcleo arqueado coexpressa a dinorfina e neuroquinina B, que exercem uma ação autorreguladora sobre a secreção de kisspeptina. Considerando que a leptina, a kisspeptina, neuroquinina B e dinorfina são apontados como reguladores do início da puberdade e da função reprodutiva, este trabalho investigou como a dieta hiperlípidica (HFD) pode influenciar esses moduladores ao longo do início do desenvolvimento puberal e após a maturação sexual. Camundongos fêmeas Kiss1-hrGFP foram tratadas com a HFD ou dieta controle a partir do desmame e foram decapitadas aos 28, 32 e 63 dias (Diestro II) e o sangue coletado para dosagem plasmática de leptina, LH e estradiol. Um segundo grupo de fêmeas, tratadas com HFD ou dieta controle, receberam com 32 e 63 dias (Diestro I) uma injeção intraperitoneal de leptina (0,5 mg/kg) ou de veículo, 45 minutos após a injeção foram anestesiadas e submetidas à perfusão cerebral. Cortes hipotalâmicos foram utilizados para realização da técnica de imunoistoquímica para dupla marcação: Kiss1-p-STAT3/ p-STAT3-CART/ Kiss1-Dyn e Kiss1-NKB. Verificamos que a HFD administrada após o desmame, quando comparada à dieta controle, antecipou a abertura vaginal, o primeiro estro e o ínicio da ciclicidade em fêmeas Kiss1-hrGFP. Como indicativo de ativação do eixo reprodutivo neste grupo, foi observado aumento do peso do útero e ovário, aos 32 dias. Esses efeitos estiveram associados com: a) o aumento da concentração plasmática de leptina; b) aumento da ativação de p-STAT3 no núcleo arqueado (ARC) e c) aumento de p-STAT3 em neurônios Kiss1-hrGFP e em neurônios CART do ARC em resposta ao estímulo da leptina. A HFD não afetou a colocalização entre Kiss1-Dyn e Kiss1-NKB em fêmeas no início do desenvolvimento puberal. Em fêmeas Kiss1-hrGFP (63 dias) a administração da HFD por 6 semanas não alterou a regularidade do ciclo estral, houve elevação da leptina plasmática associada com diminuição da expressão da p-STAT3 no ARC em resposta ao estímulo com leptina, sem alterar a responsividade dos neurônios Kiss1-hrGFP a este hormônio. Em conclusão, 9 os resultados demonstram que a antecipação da puberdade induzida pela HFD está associada com o aumento da ação central da leptina, que ao estimular o sistema kisspeptidérgico, direta ou indiretamente, por meio de neurônios CART, pode aumentar o tônus excitatório sobre o eixo reprodutivo. A administração da HFD após o desmame (6 semanas) promoveu uma redução à resposta da leptina no ARC na vida adulta, porém não afetou a responsividade dos neurônios Kiss1-hrGFP a este hormônio e nem a regularidade do ciclo estral nas fêmeas Kiss1-hrGFP. Em conjunto, esses resultados indicam que as ações da leptina nesses neurônios foram preservadas, mesmo com a resistência parcial de suas ações no ARC. Estudos adicionais deverão ser realizados para se avaliar os efeitos a longo prazo da HFD sobre o eixo reprodutivo. / The increase of childhood obesity has been correlated with the early onset of puberty. In prepubertal animals it appears high fat diet (HFD) induces anticipation of puberty. Several studies have indicated that the effect of leptin on reproductive axis can be mediated by its action on Kiss1 neurons in the hypothalamus.A subpopulation of Kiss1 neurons in the arcuate nucleus also shows co-expression of neurokinin B and dynorphin A,presenting a self-regulatory action kisspeptin secretion. Since leptin, kisspeptin, neurokinin B and dynorphin are identified as regulators of puberty onset and reproductive function, this study investigated the influence of high fat diet on these modulators along puberty onset and after sexual maturation. A group of Kiss1-hrGFP-female mice received the HFD or regular chow after weaning. The Kiss1-hrGFP females were decapitated at 28, 32 and 63 days old (Diestrus II) and blood was collected for measurement of plasma leptin, LH and estradiol. A second group of females, treated with HFD or chow after weaning, received at 32 and 63 days old (Diestrus I) an intraperitoneal injection of leptin (0.5 mg / kg) or vehicle, and 45 minutes after they were anesthetized and subjected to cerebral perfusion. Immunohistochemistry was performed for double labeling: Kiss1-p-STAT3/ p-STAT3-CART/ Kiss1-Dyn and Kiss1-NKB in the hypothalamus. It was found that, compared to controls; the HFD anticipated the vaginal opening, the first estrus and the beginning of cyclicity in Kiss1-hrGFP females. An increase in the uterine weight and ovaries, confirmed the activation of reproductive axis in the HFD group at 32 days old. These effects were associated with: a) an increase in plasma leptin concentration; b) the activation of p-STAT3 in the arcuate nucleus (ARC) and c) the increase of p-STAT3 in Kiss1-hrGFP neurons and in CART neurons of the ARC in response to the stimulus of leptin. HFD didnt affect colocalization between Kiss1-Dyn and Kiss1-NKB in females mice at hte pubertal onset . Kiss1-hrGFP female mice (63 days old), exposed to HFD for 6 weeks,exhibited regular estrous cycles, increased plasma levels of leptin associated with a decreased p-STAT3 expression in the ARC after leptin stimulation, with no changes in the responsiveness of Kiss1-hrGFP neurons to this hormone, compared to controls. In conclusion, 11 the results demonstrate that the early puberty induced by the HFD is associated with increased central leptin action, by stimulating the kisspeptidergic system, directly or indirectly, through CART neurons that can increase the excitatory tone in the reproductive axis. The administration of HFD after weaning (6 weeks) induced a decrease on the response to leptin in the ARC in adulthood, but without changes in the responsiveness of Kiss1-hrGFP neurons to this hormone and the regularity of the estrous cycle in Kiss1-hrGFP females. Taken together, these results indicate that the actions of leptin in kisspeptin neurons were preserved after 6 weeks of HFD, despite the partial resistance to this hormone in the ARC. Additional studies should be conducted to evaluate the long-term effects of HFD in the reproductive axis.

La reproduction est intimement liée à la balance énergétique, particulièrement chez les espèces sauvages exposées à des variations larges de leur environnement. L’objectif de cette thèse était d’étudier les mécanismes centraux qui régissent la régulation saisonnière de ces deux fonctions chez la gerboise. Nos résultats montrent une augmentation coordonnée au printemps de l’expression des gènes codant pour les neuropeptides impliqués dans la régulation de la reproduction et de la prise alimentaire, et rapportent des effets opposés de deux peptides classiquement considérés comme régulant la reproduction, Kisspeptine et RFRP-3, sur la prise alimentaire de la gerboise femelle ; un effet inhibiteur de Kisspeptine ayant lieu uniquement au printemps, et un effet activateur de RFRP-3 s’observant pendant les deux saisons. De plus, nous proposons que Kisspeptine et RFRP-3 exercent leurs effets sur la prise alimentaire via des actions sur des structures cérébrales dédiées au contrôle métabolique, notamment POMC et NPY. Dans l’ensemble, ces résultats renforcent notre hypothèse d’une coordination centrale de l’activité de la reproduction et de la prise alimentaire chez la gerboise et suggèrent une modulation de cette coordination en fonction du sexe et de l’environnement saisonnier. / Reproduction is intimately related to energy balance, especially in wild species exposed to marked seasonal changes in their environment. The aim of this thesis was to study the central mechanisms governing the seasonal regulation of these two functions in the jerboa. Our results reveal a spring coordinated increase in the expression of genes encoding neuropeptides involved in the regulation of reproduction and food intake, and report opposite effects of two central regulators of reproduction, Kisspeptin and RFRP-3, on food intake in the female jerboa; an inhibitory effect of Kisspeptin that occurs only in spring, and activatory effect of RFRP-3 observed in both seasons. Moreover, we propose that Kisspeptin and RFRP-3 display their effects on food intake via actions on brain structures dedicated to metabolic regulation, mainly POMC and NPY. Overall, these results strengthen our hypothesis of a central coordination of the jerboa’s reproductive activity and food intake and suggest a modulation of this coordination that depends on sex and seasonal environment.

No description available.

Aquaculture is an essential developing sector for world food production, however the attainment of sexual maturity during commercial on-growing is a major bottleneck to industry expansion. Sexual maturation brings a commercial loss due to reduced growth performance as well as reduced immune function. Furthermore, serious concerns exist over potential genetic interaction with native stocks through broadcast spawning or spawning interaction by escapees. In the north Atlantic region, the Atlantic cod (Gadus morhua) and Atlantic halibut (Hippoglossus hippoglossus) are key aquaculture species in which industry expansion is limited by pre-harvest sexual maturation. However, through a species specific combination of modern technologies and refinement in management practices it is possible that this sexual maturation can be controlled and on-growing potential enhanced. Thus the overall aim of this thesis was to conduct novel research that will improve our understanding of the underlying mechanisms that regulate sexual maturation, whilst also advancing the optimisation of technologies for the management of maturation in cod and halibut. In Atlantic cod, owing to the inconsistent inhibition of maturation in commercial conditions, ever increasing intensities of light and in some cases narrow spectrum technologies are being used to try to combat this problem. Firstly, this PhD project investigated the potential welfare impacts of high intensity artificial lighting which have not been studied to date (Chapter 2). The work specifically investigated the effect of traditional metal halide and novel green cathode lighting on the stress response, innate immunity, retina structure, feeding activity and light perception of Atlantic cod. Results indicated that although acute responses to light were observed, there were no clear significant long term effects of any of the lighting treatments on these parameters. Regarding light perception, interestingly even when subjected to high intensity constant lighting (metal halide mean tank intensity: 16.6 watts m-2), cod still demonstrated a day/night rhythm in melatonin release which suggests perception of the overlying ambient photoperiod. The second trial of this PhD project investigated the efficacy of shading of ambient photoperiod in addition to constant lighting to inhibit maturation of cod outdoors (Chapter 3). This aimed at improving the performance of artificial lighting regimes in the open cage system during commercial on-growing by reducing the relative difference between day/night light intensities. The trial was conducted over a one year period where a low and high shade treatment were tested in outdoor tanks. Shading increased the relative night time illumination to 6.6% and 31.3% of daytime levels respectively, compared to <2% in an unshaded set-up. Both shading treatments were effective at suppressing sexual development in cod as confirmed through measurements of gonadosomatic index, histological analysis of gonadal development, oocyte diameter measurements and sex steroid profiles as well as measurements of growth. In addition to research at the applied level in Atlantic cod, this thesis has also extended to the fundamental level and explored one of the potential mechanisms relaying photoperiod signal to the endogenous regulation of sexual maturation in cod, namely the kisspeptin system (Chapter 4). Partial sequences for the signal peptide Kiss2 and its receptor Kissr4 were isolated and described showing similarity to other teleost species such as the medaka, Oryzias latipes and stickleback, Danio rerio. Novel molecular qPCR assays were designed and developed to measure the expression of both genes in male and female cod over a maturation cycle and compared to cod under constant lighting which remained immature. Interestingly, expression patterns of kiss2 and kissr4 did not reveal any clear association with season or photoperiod treatment. However, pituitary expression of gonadotropins (FSH, follicle stimulating hormone; LH, luteinising hormone) did show a differential expression in relation to treatment from early winter approximately 4-6 months after the photoperiod change. These new results are in contradiction with the hypothesis that the kisspeptin system would be involved in the initiation of gametogenesis, as shown in mammals. However, the FSH/LH data defines a window during which time kisspeptin or another GnRH stimulating mechanism must be active, this compels the need further investigation. In Atlantic halibut farming, all-female production removes the concerns of production losses through sexual maturation. Accordingly, this thesis investigated the potential/feasibility of generating monosex populations by FACS (fluorescence activated cell sorting) semen sexing based on cellular DNA content, as proven in terrestrial agriculture. Results however did not show any clear differences between the DNA of sperm in a range of species tested (Atlantic halibut, cod, sea bass, perch) suggesting that this technique may not be applicable in such species. The project also focussed on the production of a population of sex reversed halibut broodstock (neomales) that will generate, in the long term, a basis for traditional monosex population generation in the UK. Two in feed MDHT (17α-methyldihydrotestosterone) treatments were tested with the aim to reduce the use of hormone. Results were very successful with a hormone treatment of 5ppm MDHT generating a 97% phenotypic male population thus suggesting the presence of sex-reversed halibut which can be used for future monosex production. Overall, this work aimed to develop and/or refine potential remediation techniques for sexual maturation in two key commercially important farmed marine fish species, cod and halibut, as well as further our understanding on the regulation of puberty. The knowledge gained from this work provides a means to optimise the techniques employed in the industry and has the potential to increase production and profitability without compromising farmed animal welfare, thus ultimately promoting the sustainable expansion of the Atlantic cod and halibut aquaculture.

No description available.

Chez la souris, la puberté de la femelle est accélérée par des phéromones urinaires émises par le mâle (effet Vandenbergh). Les mécanismes neuroendocriniens sous-Jacents et les conséquences comportementales restent peu connus. Par une approche multidisciplinaire alliant immunohistochimie, chromatographie gazeuse couplée à la spectrométrie de masse et chirurgie expérimentale, mon travail de thèse montre que les neurones synthétisant la kisspeptine, un neuropeptide hypothalamique jouant un rôle essentiel dans le contrôle de la puberté, sont régulés positivement par les phéromones accélératrices de la puberté. Les neurones à kisspeptine reçoivent le signal phéromonal via le système olfactif accessoire et le transmettent aux neurones à GnRH. De plus, des analyses comportementales montrent qu’outre leur effet physiologique connu, les phéromones accélératrices de la puberté modifient à long terme le comportement sexuel de la souris femelle. En effet, la préférence de la femelle pour l’odeur du mâle s’exprime plus tôt à l’âge adulte après l’exposition péripubère aux phéromones émises par la souris mâle. / In the mouse, female puberty onset is accelerated by male urinary pheromones (Vandenbergh effect). The neuroendocrine mechanisms underlining this effect and the behavioral consequences are poorly understood. Through a multidisciplinary approach using immunohistochemistry, gas chromatography coupled to mass spectrometry and experimental surgery, my thesis research show that neurons that synthesize kisspeptin, a hypothalamic neuropeptide which plays a master role in the control of puberty onset, are positively regulated by puberty-Accelerating pheromones. Kisspeptin neurons receive pheromone signal via the accessory olfactory system and transmit it to GnRH neurons. Moreover, behavioral analyses show that besides their known physiological effect, puberty-Accelerating pheromones also have long-Term effects on sexual behavior of the female mouse. Indeed, puberty-Accelerating pheromones induce a precocious expression of male-Directed odor preference in adult female mice.