Alors que le protéome de l'ovocyte isolé, aux stades VG et MII a déjà été étudié, celui du follicule en croissance n'a jamais été décrit.Dans cette étude, nous avons cherché à identifier, comparer et caractériser les profils protéiques de follicules ovariens de souris à trois stades de leur développement in vitro distincts morphologiquement : follicules secondaires en début de culture - stade initial (IS), follicules avec une rupture complète de la membrane de Slavjanski (RMS) et follicules avec une cavité similaire à l'antrum (FA).Après un préfractionnement par IEF et une analyse LC-MS/MS en deux configurations (1D et 2D), 1403 protéines ont pu être identifiées dans le follicule ovarien de souris. 43,4 % (609) des protéines identifiées étaient communes aux trois stades et d'autres ont été identifiées uniquement à un stade : 71 au stade IS, 182 au stade RMS et 193 au stade FA. De plus, on a identifié 365 protéines qui n'avaient pas été décrites antérieurement dans le protéome de l'ovocyte ce qui pourrais indiquer qu'elles sont exprimées dans les cellules somatiques du follicule. Des analyses qualitatives et quantitatives complémentaires ont démontré une surreprésentation pour 44 fonctions biologiques par rapport aux fonctions biologiques des gènes constituant le génome de Mus musculus et mis en évidence des différences d'expression et d'abondance des protéines liées au cycle cellulaire, à la fixation des ions de calcium et à la glycolyse selon le stade de développement. Ces résultats représentent un point de départ pour beaucoup d'autres études de caractérisation moléculaire du développement folliculaire. / Until now only the proteome of isolated oocyte from fully grown follicle were described with the aim of oocyte maturation characterization. However the ability of oocyte to mature is acquired during its development within the follicle. Thus in this study we proposed a protein identification and characterization of whole mice ovarian follicle at three morphological stages during in vitro development: early secondary stage, described as initial stage (IS), follicles with a complete Slavjanski membrane rupture (RMS) and follicles with an antrum like cavity formation (FA). Using an IEF pre fractionation before protein digestion and two configurations of LC-MS/MS analysis (1D and 2D), 1403 proteins were successfully identified in the murine ovarian follicle. From those, 43.4 % (609) were commonly identified in the three stages and some were identified only at one single stage: 71 at IS stage, 182 at RMS stage and 193 at FA stage. Compared to the proteomes of isolated oocyte previously described, 365 proteins were only identified in our samples indicating that those ones were probably expressed in the somatic cells of the follicle. Additional qualitative and quantitative analysis highlighted 44 biological processes over represented in our samples when compared to Mus musculus gene database and different expressions and protein abundance implicated in cell cycle, calcium ion binding and glycolysis, throughout in vitro follicle development. This report represents so far the most complete catalogue of follicle proteins and could be an important milestone in the proteomic study of the follicle metabolism throughout in vitro development.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-06-01T11:44:36Z No. of bitstreams: 1 ralphmaturanopinheiro.pdf: 530518 bytes, checksum: c7705e2c22719ac685677b3a3ed3fe26 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-07-02T12:54:45Z (GMT) No. of bitstreams: 1 ralphmaturanopinheiro.pdf: 530518 bytes, checksum: c7705e2c22719ac685677b3a3ed3fe26 (MD5) / Made available in DSpace on 2016-07-02T12:54:45Z (GMT). No. of bitstreams: 1 ralphmaturanopinheiro.pdf: 530518 bytes, checksum: c7705e2c22719ac685677b3a3ed3fe26 (MD5) Previous issue date: 2012-02-03 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A bionomia de Chelopistes meleagridis fora do hospedeiro foi observada com o objetivo de compreender aspectos relacionados ao ciclo de vida desta espécie. Para isto, adultos de C. meleagridis foram coletados e colocados em condições controladas para se reproduzir, oferecendo-se pena como alimento. Da prole destes adultos, foi observado o desenvolvimento de 150 indivíduos desde o ovo até a fase adulta. Para 75 destes, foi oferecida a dieta composta de pena, enquanto para os outros 75 a dieta foi composta de pena e pele do hospedeiro (peru, Meleagris gallopavo). Ao verificar que a dieta “pena + pele” foi a que resultou no maior número de adultos, foram observadas a fertilidade, fecundidade e a longevidade de piolhos criados in vitro desde o primeiro ínstar alimentados com esta dieta. Valores altos relacionados à reprodução desta espécie foram encontrados em relação a outros piolhos da subordem Ischnocera, destacando-se: número de ovos produzidos por dia e número de ovos produzidos por fêmeas durante a vida (médias de 2,54 e 26,61 ovos, respectivamente, para fêmeas selvagens e 2,11 e 29,33 ovos, respectivamente, para fêmeas criadas in vitro.). A inclusão de pele na dieta foi determinante para o desenvolvimento até o estágio adulto, uma vez que 48% dos piolhos alimentados com essa dieta atingiram a fase adulta. Quando foi oferecido apenas pena, 1,3% dos piolhos atingiram a maturidade. O tempo de desenvolvimento de machos e fêmeas foi semelhante (média de 29,38 dias) sem haver diferença na proporção sexual dos adultos. / Bionomics of the large louse turkey Chelopistes meleagridis off host was evaluated to comprehend aspects related to life cycle. To this, adults was collected and put in controlled conditions for reproduce, offered feathers as food. From offspring of these adults were observed the development of 150 lice from eggs to adults. To 75 of them was given only feathers as resource of food while to others 75 lice the diet was composed by feathers and skin debris from the natural host, Meleagris gallopavo. When verified that diet composed by feather + skin resulted in more adults, was evaluated fertility, fecundity and longevity of this specie reared in vitro from first nymphs to adults feed with this diet. High values related to reproduction of this specie were found in comparison with others avian Ischnocera, like: number of eggs produced by day and number of egg produced during the lifespan of females (means of 2.54 and 26.61, respectively to wild females and 2.11 and 29.33 respectively to females reared in vitro). The addition of skin in diet was determinant to development to adult stage, once time 48% fed with this diet reared to adults while the lice feed with feathers only 1.3 % of then became adult stage. The time of development of males and females was the same (29 days). The sex rate of adults reared in vitro was 1:1.