If sexual selection acts in a sex-limited way on a trait that has a shared genetic basis between males and females, the resulting intralocus sexual conflict can have a considerable impact on the opposite sex. A prime example of sexual conflict affecting a shared characteristic would be the reproductive organs in males and females. This study investigates how artificial selection on the female-specific trait egg size influences male reproductive success, particularly male testes morphology, by analysing the density and number of seminiferous tubules within the testes. It was found that selection on female reproductive investment has a concordant effect on male reproductive success, by demonstrating increased density of seminiferous tissue in the testes of males originating from high investment selection lines. Interestingly, a non-significant trend suggested that the density of seminiferous tissue influences testes size in a negative way. This study therefore provides evidence that female-specific selection on reproductive investment influences testes morphology in males, and that testes size depends on more than sperm-producing tissue.

No description available.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21008号 / 医博第4354号 / 新制||医||1028(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 長船 健二, 教授 小川 修, 教授 横出 正之 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 新制・課程博士 / 博士(医学) / 甲第25004号 / 医博第5038号 / 新制||医||1070(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 長船 健二, 教授 生田 宏一, 教授 上野 英樹 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Les glandes salivaires sont des glandes mixtes : la salive (produit exocrine) estimpliquée dans le maintien de l’homéostasie buccale alors que les secrétions endocrines (ex :facteurs de croissance) ont un rôle physiologique (gamétogénèse, ostéogenèse,hypertension…) Chez les mammifères, elles affichent un dimorphisme sexuel qui laisseentrevoir une sensibilité éventuelle à des xeno-hormones.Ce mémoire présente l’action de la génistéine (phyto-oestrogène) et/ou de la vinclozoline(anti-androgène) sur la glande submandibulaire (SM) de rat lors d’une exposition précoce viala mère (gestation-lactation) et lors d’une exposition pendant la période de croissance (dusevrage à l’âge adulte). Les glandes SM, prélevées au stade immature et jeune adulte, ont faitl’objet d’une analyse histologique et d’une étude de marqueurs moléculaire des fonctionsendocrines et exocrines associées aux processus gustatifs. L’exposition précoce ralenti ledéveloppement de la glande SM et augmente sélectivement la préférence au sucré des malesimmatures mais pas des adultes ; l’analyse moléculaire révèle une action sélective sur lesfonctions exocrines corrélée à celle sur les préférences, ainsi qu’une action sur les fonctionsendocrines (facteurs de croissances) qui s’inverse avec l’âge. L’exposition à partir du sevrageperturbe seulement les mâles qui présentent des altérations des structures sécrétrices coupléesà des modifications d’expression des récepteurs hormonaux et facteurs de croissance, maisaussi au taux sérique de l’EGF.Cette étude identifie la glande submandibulaire comme cible de perturbateurs endocriniens etpose la question des conséquences physiologiques à terme / The salivary glands are mixed glands: saliva (exocrine product) is involved inmaintaining oral homeostasis whereas endocrine secretions (eg growth factors) have aphysiological role (gametogenesis, osteogenesis, hypertension ..). In mammals, they displaysexual dimorphism suggesting a possible susceptibility to xeno-hormones.This manuscript presents the action of genistein (phytoestrogen) and/or vinclozolin (antiandrogenic)on the submandibular gland (SM) rats when performing an early exposure via themother (pregnancy, lactation) or an exposure during the growth period (from weaning toadulthood). The SM glands, collected at immature and young adult ages, have been analyzedaccording histological aspect and expression of molecular markers of endocrine and exocrinefunctions associated with gustatory process. The early exposure disrupted the development ofthe SM gland and selectively increases the sweet preference in immature males but not inadults; molecular analysis reveals a selective action on exocrine functions related to the sweetpreferences and also an action on endocrine functions (growth factors) which reverses withage. Exposure from weaning disrupts only the male salivary glands with alterations insecretory structures coupled with changes in expression of both, sex-hormone receptors andgrowth factors, but also in serum EGF.This study identifies the submandibular gland as a target for endocrine disruptors and raisesthe question of the further physiological consequences

O parálogo 3 do trocador Na+/H+ (NHE3) é essencial para a reabsorção de HCO3- nos túbulos proximais renais e sua expressão e função adaptam-se às diferentes condições ácido-base do organismo. O objetivo desta tese foi avaliar quais as diferenças entre os efeitos da acidose metabólica (AM) e respiratória (AR) sobre a regulação do NHE3 e identificar variáveis responsáveis pelas respostas adaptativas observadas. Em células OKP, a AM foi simulada diminuindo a [HCO3-] do meio de cultura e a AR aumentando a pCO2 ambiente por 24 h. Foram observados os efeitos das acidoses sobre o RNAm-Nhe3, a presença da proteína-NHE3 na membrana celular e a atividade promotora do gene do Nhe3. Concluiu-se que o pH extracelular não é a variável físico-química responsável por estimular a expressão do NHE3, contudo é um importante candidato à variável responsável por regular o tráfego da proteína para a membrana. Além disso, a região de -471 a -153 pb em relação ao sítio de início de transcrição do promotor do gene do Nhe3 contém prováveis reguladores positivos que atuam em resposta à AM. / The Na+/H+ exchanger 3 (NHE3) is essential for HCO3- reabsorption in renal proximal tubules and its expression and function must adapt to acid-base conditions. The goal of the presente study was to evaluate whether there are differences between metabolic (MA) and respiratory acidosis (RA) with regard to NHE3 modulation and to identify variables that may trigger these distinct adaptive responses. In OKP cells, MA was achieved by lowering [HCO3-] in the cell culture medium and RA by increasing pCO2 in the incubator chamber for 24 h. The effects of both acidosis on Nhe3 mRNA levels, cell-surface NHE3 expression and promoter activity were evaluated. In summary, it was concluded that low extracellular pH is not the physical-chemical variable that up-regulates NHE3 expression, however, extracellular pH is a candidate for the variable related to the NHE3 displacement to the apical membrane. Moreover, the Nhe3 gene promoter region spanning from -471 to -153 base pairs upstream from the transcriptional start site contains putative enhancers regulated in response to MA.

Le syndrome de Bartter de type III résulte de mutations du gène CLCNKB codant le canal chlorure CLC-KB. Un grand nombre de mutations a été répertorié, mais peu d'entre elles ont été caractérisées fonctionnellement. Devant ce manque de données, nous nous sommes fixés comme objectif de procéder à une analyse fonctionnelle des mutations de CLCNKB pour tenter d'approfondir davantage les mécanismes de régulation de CLC-KB. Ce travail a nécessité l'emploi de trois systèmes d'expression hétérologue, les ovocytes de Xenopus laevis et les lignées cellulaires rénales HEK293T et MDCK, afin de procéder à des analyses électrophysiologiques et des expériences de biologie cellulaire. Nous démontrons que toutes les mutations étudiées altèrent l'expression membranaire de la protéine et que la conductance résiduelle est proportionnée à cette expression. Ainsi, la réduction des courants provient d'une réduction du nombre de canaux et non d'une altération majeure de la conduction ou de la régulation. Nous rapportons également que certaines mutations modifient la sensibilité au pH et au calcium extracellulaires du CLC-KB. Cette altération peut avoir un impact pathologique majeur. En conclusion, cette étude fonctionnelle montre le rôle essentiel du CLC-KB dans le maintien de la balance sodée et laisse entrevoir un espoir thérapeutique à long terme de restauration des anomalies précisément identifiées. Enfin, l'analyse de mutations est un outil extrêmement puissant pour les études structure-fonction. Associée à la recherche de nouveaux partenaires, elle devrait permettre de découvrir les régulations physiologiques de ce canal, pour l'heure presqu'inconnues.

Made available in DSpace on 2015-03-26T13:47:30Z (GMT). No. of bitstreams: 1 texto completo.pdf: 256877 bytes, checksum: d548a53fcd07f9ed376af138ca9245bb (MD5) Previous issue date: 2007-07-31 / The crab-eating fox (Cerdocyon thous), known as graxaim, lobinho, etc. belongs to the order Carnivora and family Canidae, being found at the most part of Brazil. This animal does not figure in the list of IBAMA s (Brazilian Institute of Environment and Renewable Natural Resources, 2007) threatened animals of extinguishing and even relatively abundant, little is known about its social, morphological and physiological biology, in special those related with the basic fundamentals of its reproductive physiology. The study of basic aspects of reproduction, in special the spermatogenic process, is relevant to provide informations for the development of technologies in attended reproduction. The quantitative analysis of spermatogenesis can be used to establish kinetic patterns of division and renewal of spermatogonias. It allows estimate the efficiency of the spermatogenic process, the quantification of germ cells population and its relationship with the Sertoli cells population, in some stages of the spermatogenic process development. The present work defines some testicular volumetric parameters and correlates the morphometry of testis and seminiferous tubule data with corporal weight. It also characterizes and classifies the intertubular space of crab-eating fox. In this way, it had been used six adults crab-eating fox, from the Wild Animals Selection Center (CETAS-UFV). To collect samples was chosen the testis biopsy, which is an appropriate methodology for sample collection for the study of the physiologic and reproductive pathologies in domestic and wild animals, as it provides sufficient material for histological analysis, not presenting any alteration in long term spermatogenic process. In this work, the average body weight of the crab-eating fox was 6.53 kg, of which 0.068% are allocated in testicular mass and 0.042% specifically in seminiferous tubules, which represented 87.5% of testicular parenchyma. The testicular albuginea represented 12.5% of testicular mass. The average diameter of seminiferous tubules was 236 μm, the average epithelium thickness was 629 μm, with 18 meters of seminiferous tubule per gram of testis. In relation to the testicular parenchyma, 3.88% is occupied by Leydig cells, 7.95% by conjunctive tissue, 0.12% by lymphatic vessels and 0.72% by blood vessels. The average volume of Leydig cell was 1149 μm3 and the average nuclear diameter was 8.01 μm. In crab-eating fox the Leydig cells are on average 0.0022% of body weight and its average number per gram of testis (31 million) is within the expected for mammalians, between 20 and 40 million. / O cachorro-do-mato (Cerdocyon thous), conhecido como graxaim, raposacaranguejeira, lobinho, lobete e graxaim-do-mato, pertence à ordem Carnivora e família Canidae, sendo encontrado na maior parte do território brasileiro. Este animal não figura na lista de animais ameaçados de extinção do IBAMA (Instituto Brasileiro de Meio Ambiente e dos Recursos Naturais Renováveis, 2007) e embora relativamente abundante, pouco é conhecido sobre sua biologia social e morfofisiológica, em especial quanto aos fundamentos básicos da sua fisiologia reprodutiva. O estudo de aspectos básicos da reprodução, em especial o processo espermatogênico, torna-se relevante para fornecer informações para o desenvolvimento de tecnologias de reprodução assistida. A análise quantitativa da espermatogênese pode ser utilizada no estabelecimento de padrões cinéticos, como a divisão e renovação de espermatogônias. Permite ainda estimar o coeficiente de eficiência do processo espermatogênico e quantificar a população das células germinativas e suas relações com a população das células de Sertoli em várias etapas do desenvolvimento ao longo do processo espermatogênico. O presente trabalho se propôs a definir alguns parâmetros volumétricos testiculares e correlacionar os dados de morfometria testicular e do túbulo seminífero à massa corporal, além de caracterizar e classificar o espaço intertubular do cachorro-domato. Foram utilizados seis cachorros-do-mato adultos, provenientes do Centro de Triagem de Animais Silvestres CETAS-UFV. Para a coleta das amostras foi realizada a biópsia testicular, que é uma metodologia eletiva para a coleta de material para o estudo da fisiologia e patologias reprodutivas em animais domésticos e selvagens, uma vez que fornece material suficiente para análise histológica, não apresentando quaisquer alterações sobre o processo espermatogênico em longo prazo. No presente trabalho, o peso corporal médio dos cachorros-do-mato foi de 6,53 Kg, dos quais cerca de 0,068% estão alocados em massa testicular e 0,042% especificamente em túbulos seminíferos, que representaram cerca de 87,5% do parênquima testicular .A albugínea testicular representou cerca de 12,5% da massa testicular. O diâmetro médio dos túbulos seminíferos foi de 236 μm, e a espessura média do epitélio seminífero foi de aproximadamente 62,9 μm, apresentando 18 metros de túbulo seminífero por grama de testículo. Com relação ao parênquima testicular, 3,88% é ocupado por células de Leydig, 7,95% por tecido conjuntivo, 0,12% por vasos linfáticos e 0,72% por vasos sangüíneos. O volume médio da célula de Leydig foi de 1149 μm3 e o seu diâmetro nuclear médio foi de 8,01 μm. Em cachorros-do-mato, as células de Leydig ocupam em média 0,0022% do peso corporal, com o número médio dessas células de 31 milhões por grama de testículo, que está dentro da amplitude descrita para a maioria dos mamíferos, entre 20 e 40 milhões.

O parálogo 3 do trocador Na+/H+ (NHE3) é essencial para a reabsorção de HCO3- nos túbulos proximais renais e sua expressão e função adaptam-se às diferentes condições ácido-base do organismo. O objetivo desta tese foi avaliar quais as diferenças entre os efeitos da acidose metabólica (AM) e respiratória (AR) sobre a regulação do NHE3 e identificar variáveis responsáveis pelas respostas adaptativas observadas. Em células OKP, a AM foi simulada diminuindo a [HCO3-] do meio de cultura e a AR aumentando a pCO2 ambiente por 24 h. Foram observados os efeitos das acidoses sobre o RNAm-Nhe3, a presença da proteína-NHE3 na membrana celular e a atividade promotora do gene do Nhe3. Concluiu-se que o pH extracelular não é a variável físico-química responsável por estimular a expressão do NHE3, contudo é um importante candidato à variável responsável por regular o tráfego da proteína para a membrana. Além disso, a região de -471 a -153 pb em relação ao sítio de início de transcrição do promotor do gene do Nhe3 contém prováveis reguladores positivos que atuam em resposta à AM. / The Na+/H+ exchanger 3 (NHE3) is essential for HCO3- reabsorption in renal proximal tubules and its expression and function must adapt to acid-base conditions. The goal of the presente study was to evaluate whether there are differences between metabolic (MA) and respiratory acidosis (RA) with regard to NHE3 modulation and to identify variables that may trigger these distinct adaptive responses. In OKP cells, MA was achieved by lowering [HCO3-] in the cell culture medium and RA by increasing pCO2 in the incubator chamber for 24 h. The effects of both acidosis on Nhe3 mRNA levels, cell-surface NHE3 expression and promoter activity were evaluated. In summary, it was concluded that low extracellular pH is not the physical-chemical variable that up-regulates NHE3 expression, however, extracellular pH is a candidate for the variable related to the NHE3 displacement to the apical membrane. Moreover, the Nhe3 gene promoter region spanning from -471 to -153 base pairs upstream from the transcriptional start site contains putative enhancers regulated in response to MA.

Current in vitro cellular methods utilised in drug metabolism and pharmacokinetic (DMPK) studies during drug development do not provide the 3D structure and functions of organs found in vivo, such that resulting in vitro-in vivo extrapolation (IVIVE) may not always accurately reflect clinical outcome. This highlights the need for the development of new dynamic in vitro cell models to aid improvement of IVIVE. The aim of this project was to incorporate characterised primary renal cells within a hollow fibre bioreactor for use in DMPK studies investigating renal clearance. Fluorescence based assays were developed to assess the functionality of three drug transporters involved in the renal transport of pharmaceutical compounds: P-gp, BCRP and OCT2. The developed assays were then applied alongside transporter visualisation and genetic expression assays to characterise primary human proximal tubule cells over a series of population doublings. Cells at a population doubling of 5 demonstrated the best transporter activity whilst allowing cells to be expanded in vitro. Polysulfone (PSF) based membranes, which are widely used in dialysis components were developed by blending additives to improve renal cell attachment and culture. The membranes exhibited a characteristic porous internal structure with smooth skin layers on the surface, and were able to be sterilised via autoclaving due to their high thermal stability. PSF blended with polyvinylpyrrolidone (PVP) was the most hydrophilic with cell metabolic activity similar to standard tissue culture plastic. The production of hollow fibres of varying thicknesses and properties from the PSF and PVP blend yielded a marked difference in renal cell attachment and long term viability. Fibres incorporated into glass casings to produce the single hollow fibre bioreactors (HFBs) were able to be sterilised by autoclaving whilst remaining intact. Due to the variation of fibre integrity within the batch, many fibres exhibited tears within the HFBs. This ultimately led to cell depletion within the fibre over the culture period; however, intact fibres demonstrated an increase in cell growth towards the end of the culture period under flow conditions. These results demonstrate the progress made towards a small scale in vitro renal model incorporating characterised primary renal cells to aid the improvement of IVIVE in DMPK research.