The cosmic rays are the most energetic particles in the universe. Their production, propagation, and detection are objects of studies. Surface detectors aim to identify particles from extensive air showers (EAS) which the result from the cosmic-ray interactions with the atmosphere. Resistive Plate Chambers (RPCs) have shown to be a suitable muon detector to be integrated into the Pierre Auger Observatory. An instrumentation was developed to assembly RPCs in São Carlos (BRA). Data from RPCs already built by our collaborators in Coimbra (POR) were analyzed. The detector efficiency to muons was calculated and is approximately 88%, which is in good agreement with the values quoted in the literature. Direction maps were built to investigate the muon incoming direction and the quantity of matter traversed by the muons. The dependence of the muon flux on the zenith angle was calculated and compared with results from the simulation. A square cosine dependence is expected, though it is seen that the building structure has enough matter to block some of the incident muons and alter the dependence curve. The total muon flux was estimated based on the detector efficiencies and solid angle as 1.6.10−5. mm−2.sr−1. s−1 compared with the literature value of 7.1.10−5 mm−2.sr−1.s−1, which gives an absorption by the building of approximately 77%. / Os raios cósmicos são as partículas mais energéticas do universo. Sua produção, propagação e detecção são objetos de estudos. Os detectores de superfície têm como objetivo identificar partículas dos chuveiros atmosféricos extensos (EAS), o qual é o resultado das interações do raio cósmico com a atmosfera. A Câmaras de Placas Resistivas (RPCs) demonstra ser um detector de múons adequado para ser integrado ao Observatório Pierre Auger. Foi desenvolvida em São Carlos (BRA) uma instrumentação para montagem de RPCs. Dados de RPCs já construídas por nossos colaboradores em Coimbra (POR) foram analisados. A eficiência dos detectores para múons foi calculada como sendo de aproximadamente 88%, o que está de acordo com os valores citados na literatura. Mapas de direção foram construídos para investigar a direção de chegada e a quantidade de matéria atravessada pelos múons. A dependência do fluxo de múons com o ângulo zenital foi comparada com os resultados da simulação. Embora uma dependência com o quadrado do cosseno é esperada, foi constatado que a estrutura do prédio tem matéria suficiente para bloquear parte dos múons incidentes e alterar a curva da dependência. O fluxo total de múons foi estimado baseado nas eficiências do detector e no ângulo sólido é de 1.6.10−5 mm−2.sr−1.s−1. Comparado com o valor da literatura de 7.1.10−5 mm−2.sr−1.s−1 resulta em uma absorção pelo prédio de aproximadamente 77% do fluxo de múons.

The cosmic rays are the most energetic particles in the universe. Their production, propagation, and detection are objects of studies. Surface detectors aim to identify particles from extensive air showers (EAS) which the result from the cosmic-ray interactions with the atmosphere. Resistive Plate Chambers (RPCs) have shown to be a suitable muon detector to be integrated into the Pierre Auger Observatory. An instrumentation was developed to assembly RPCs in São Carlos (BRA). Data from RPCs already built by our collaborators in Coimbra (POR) were analyzed. The detector efficiency to muons was calculated and is approximately 88%, which is in good agreement with the values quoted in the literature. Direction maps were built to investigate the muon incoming direction and the quantity of matter traversed by the muons. The dependence of the muon flux on the zenith angle was calculated and compared with results from the simulation. A square cosine dependence is expected, though it is seen that the building structure has enough matter to block some of the incident muons and alter the dependence curve. The total muon flux was estimated based on the detector efficiencies and solid angle as 1.6.10−5. mm−2.sr−1. s−1 compared with the literature value of 7.1.10−5 mm−2.sr−1.s−1, which gives an absorption by the building of approximately 77%. / Os raios cósmicos são as partículas mais energéticas do universo. Sua produção, propagação e detecção são objetos de estudos. Os detectores de superfície têm como objetivo identificar partículas dos chuveiros atmosféricos extensos (EAS), o qual é o resultado das interações do raio cósmico com a atmosfera. A Câmaras de Placas Resistivas (RPCs) demonstra ser um detector de múons adequado para ser integrado ao Observatório Pierre Auger. Foi desenvolvida em São Carlos (BRA) uma instrumentação para montagem de RPCs. Dados de RPCs já construídas por nossos colaboradores em Coimbra (POR) foram analisados. A eficiência dos detectores para múons foi calculada como sendo de aproximadamente 88%, o que está de acordo com os valores citados na literatura. Mapas de direção foram construídos para investigar a direção de chegada e a quantidade de matéria atravessada pelos múons. A dependência do fluxo de múons com o ângulo zenital foi comparada com os resultados da simulação. Embora uma dependência com o quadrado do cosseno é esperada, foi constatado que a estrutura do prédio tem matéria suficiente para bloquear parte dos múons incidentes e alterar a curva da dependência. O fluxo total de múons foi estimado baseado nas eficiências do detector e no ângulo sólido é de 1.6.10−5 mm−2.sr−1.s−1. Comparado com o valor da literatura de 7.1.10−5 mm−2.sr−1.s−1 resulta em uma absorção pelo prédio de aproximadamente 77% do fluxo de múons.

Des études présentées dans cette thèse ont permis de démontrer que le gène du groupe Polycomb (PcG) Bmi1 est essentiel à l’auto-renouvellement des progéniteurs rétiniens immatures et pour le développement rétinien après la naissance. Ce travail illustre chez l’embryon que Bmi1 est hautement enrichie dans une sous-population de progéniteurs rétiniens exprimant le marqueur de surface SSEA-1 et différents marqueurs de cellules souches. À tous les stades de développement analysés, l’absence de Bmi1 résulte en une diminution de la prolifération et de l’auto-renouvellement des progéniteurs immatures. Pour mieux comprendre la cascade moléculaire en absence de Bmi1, nous avons inactivé p53 dans les colonies Bmi1-/-. Cette inactivation a permis une restauration partielle du potentiel d’auto-renouvellement. De plus, en absence de Bmi1, la prolifération et la maintenance de la population de progéniteurs rétiniens immatures localisés dans le corps ciliaire sont aussi affectées après la naissance. Bmi1 permet donc de distinguer les progéniteurs immatures de la population principale de progéniteurs, et est requis pour le développement normal de la rétine. Nous avons également démontré que l’oncogène Bmi1 est requis dans les neurones pour empêcher l’apoptose et l’induction d’un programme de vieillissement prématuré, causé par une baisse des défenses anti-oxydantes. Nous avons observé dans les neurones Bmi1-/- une augmentation des niveaux de p53, de la concentration des ROS et de la sensibilité aux agents neurotoxiques. Nous avons démontré ainsi que Bmi1 contrôle les défenses anti-oxydantes dans les neurones en réprimant l’activité pro-oxydante de p53. Dans les neurones Bmi1-/-, p53 provoque la répression des gènes anti-oxydants, induisant une augmentation des niveaux de ROS. Ces résultats démontrent pour la première fois que Bmi1 joue un rôle critique dans la survie et le processus de vieillissement neuronal. / The studies presented in this thesis establish that the Polycomb Group (PcG) gene Bmi1 is required for the self-renewal of immature retinal progenitor cells (RPCs) and for postnatal retinal development. Work performed in mouse embryos reveals that Bmi1 is highly enriched in a RPC subpopulation expressing the cell surface antigen SSEA-1 and different stem cell markers. Furthermore, at all developmental stages analysed, Bmi1 deficiency resulted in reduced proliferation and self-renewal of immature RPCs. To better understand the molecular cascade leading to this phenotype, we inactivated p53 in Bmi1-deficient colonies. p53 inactivation partially restored RPCs self-renewal potential. Moreover, the proliferation and the postnatal maintenance of an immature RPC population located in the ciliary body was also impaired in absence of Bmi1. Thus, Bmi1 distinguishes immature RPCs from the main RPC population and is required for normal retinal development. We have also shown that the oncogene Bmi1 is required in neurons to prevent apoptosis and the induction of a premature aging-like program characterized by reduced antioxidant defenses. We observed in Bmi1-deficient neurons an increased p53 and ROS levels, and a hypersensitivity to neurotoxic agents. We demonstrated that Bmi1 regulate antioxidant defenses in neurons by suppressing p53 pro-oxidant activity. In Bmi1-/- neurons, p53 induces antioxidant genes repression, resulting in increased ROS levels. These findings reveal for the first time the major role of Bmi1 on neuronal survival and aging.

Des études présentées dans cette thèse ont permis de démontrer que le gène du groupe Polycomb (PcG) Bmi1 est essentiel à l’auto-renouvellement des progéniteurs rétiniens immatures et pour le développement rétinien après la naissance. Ce travail illustre chez l’embryon que Bmi1 est hautement enrichie dans une sous-population de progéniteurs rétiniens exprimant le marqueur de surface SSEA-1 et différents marqueurs de cellules souches. À tous les stades de développement analysés, l’absence de Bmi1 résulte en une diminution de la prolifération et de l’auto-renouvellement des progéniteurs immatures. Pour mieux comprendre la cascade moléculaire en absence de Bmi1, nous avons inactivé p53 dans les colonies Bmi1-/-. Cette inactivation a permis une restauration partielle du potentiel d’auto-renouvellement. De plus, en absence de Bmi1, la prolifération et la maintenance de la population de progéniteurs rétiniens immatures localisés dans le corps ciliaire sont aussi affectées après la naissance. Bmi1 permet donc de distinguer les progéniteurs immatures de la population principale de progéniteurs, et est requis pour le développement normal de la rétine. Nous avons également démontré que l’oncogène Bmi1 est requis dans les neurones pour empêcher l’apoptose et l’induction d’un programme de vieillissement prématuré, causé par une baisse des défenses anti-oxydantes. Nous avons observé dans les neurones Bmi1-/- une augmentation des niveaux de p53, de la concentration des ROS et de la sensibilité aux agents neurotoxiques. Nous avons démontré ainsi que Bmi1 contrôle les défenses anti-oxydantes dans les neurones en réprimant l’activité pro-oxydante de p53. Dans les neurones Bmi1-/-, p53 provoque la répression des gènes anti-oxydants, induisant une augmentation des niveaux de ROS. Ces résultats démontrent pour la première fois que Bmi1 joue un rôle critique dans la survie et le processus de vieillissement neuronal. / The studies presented in this thesis establish that the Polycomb Group (PcG) gene Bmi1 is required for the self-renewal of immature retinal progenitor cells (RPCs) and for postnatal retinal development. Work performed in mouse embryos reveals that Bmi1 is highly enriched in a RPC subpopulation expressing the cell surface antigen SSEA-1 and different stem cell markers. Furthermore, at all developmental stages analysed, Bmi1 deficiency resulted in reduced proliferation and self-renewal of immature RPCs. To better understand the molecular cascade leading to this phenotype, we inactivated p53 in Bmi1-deficient colonies. p53 inactivation partially restored RPCs self-renewal potential. Moreover, the proliferation and the postnatal maintenance of an immature RPC population located in the ciliary body was also impaired in absence of Bmi1. Thus, Bmi1 distinguishes immature RPCs from the main RPC population and is required for normal retinal development. We have also shown that the oncogene Bmi1 is required in neurons to prevent apoptosis and the induction of a premature aging-like program characterized by reduced antioxidant defenses. We observed in Bmi1-deficient neurons an increased p53 and ROS levels, and a hypersensitivity to neurotoxic agents. We demonstrated that Bmi1 regulate antioxidant defenses in neurons by suppressing p53 pro-oxidant activity. In Bmi1-/- neurons, p53 induces antioxidant genes repression, resulting in increased ROS levels. These findings reveal for the first time the major role of Bmi1 on neuronal survival and aging.