Navegando por Palavras-chave "Epididymal Morphogenesis"

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    Morfogênese epididimária: relevância funcional de androgênios e glicocorticoides
    (Universidade Federal de São Paulo (UNIFESP), 2020-07-30) Sousa, Maria Eleticia De [UNIFESP]; Winston, Maria Christina Werneck De Avellar [UNIFESP]; Universidade Federal de São Paulo
    The epididymis is a single and convoluted tubule, which connects the efferent ductules to the vas deferens. It is an important organ for sperm maturation, transport, storage, and their protection before ejaculation. The epididymal morphogenesis, that occurs from the mesonephric duct (its embryonic precursor), is primarily dependent on androgens by mechanisms that are not well defined yet. Recently, our research group has identified a differential expression pattern of glucocorticoids receptors (GR) on epithelial and mesenchymal cells of the developing rat mesonephric duct. The biological role of GR/glucocorticoid signaling, as well as its potential interaction with androgens during pre- and post-natal development of the epididymis is not well established yet. In the present study, we used a preclinical model of dexamethasone (synthetic glucocorticoid) antenatal treatment in rat dams (Wistar) as a tool to help to elucidate the role and function of glucocorticoids during the mesonephric duct development, with a focus on epididymal morphogenesis. Initially, we tested the daily DEX (1 mg/kg, s.c.) treament of dams during the gestational period that, in the rat, correlates with epididymal morphogenesis (e17.5-e19.5; group DEX-LW versus control, SAL-LW; euthanasia at e20.5). The specific aims, using this preclinical model, were: a) to evaluate the effects of the DEX antenatal treatment on (i) biometric and hormonal parameters in the dams, and (ii) biometric and hormonal parameters in the male fetuses, b) to characterize the effects of the DEX antenatal treatment on histomorphological parameters and cellular proliferation rate of the mesonephric duct of the male offsprings, c) to characterize the effects of the DEX antenatal treatment in the immunodistribution of AR and GR in the mesonephric duct of the male offsprings, d) to characterize the effect of the DEX antenatal treatments in the transcriptional levels of a subset of genes that are target of GR and AR regulation. Based on the results obtained, we subsequently conducted studies by exposing rat dams to daily DEX treatment during the gestacional period that involves masculinization and differntation windows (e13.5-e19.5; DEX-FL versus control SAL-FL; euthanasia at e20.5). Our readouts were: biometrical parameters (dams and fetuses), hormonal dosage (corticosterone and testosterone; dams and fetuses), histopathology, immunofluorescence studies and quantification of gene expression by RT-qPCR on isolated mesonephric duct. Althogether the results confirmed the hypothesis that glucocorticoids play a role in morphogenetic events of the mesonephric duct, including epididymal morphogenesis; also, the data contributed to the better understanding that changes in maternal plasma concentration of glucocorticoids impact the expression and function profile of GR and AR in the duct. From a translational point of view, the present study corroborates to a better understand of how corticotherapy or changes in the endougenous glucocorticoids profile during gestational period may influence reproductive health of male offsprings. (Research Ethics Committee, CEUA/UNIFESP, #8759031215).
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    Regulação Androgênica, Atividade Biológica e Sinalização Celular de Beta-Defensinas na Morfogênese do Epidídimo
    (Universidade Federal de São Paulo (UNIFESP), 2019-07-25) Ferreira, Lucas Garcia Alves [UNIFESP]; Winston, Maria Christina Werneck De Avellar [UNIFESP]; Ribeiro, Camilla Moreira [UNIFESP]; http://lattes.cnpq.br/9498472516886944; http://lattes.cnpq.br/5534065920525009; http://lattes.cnpq.br/2251799531659813; Universidade Federal de São Paulo (UNIFESP)
    Reproductive disorders in adult men, a topic of public health interest, may be related to develop-mental and masculinization abnormalities of the male reproductive tract. The mesonephric duct (or Wolffian duct) is an embryonic structure that undergoes a process of tubular-epithelial morphogenesis primarily induced by androgens that originates the epididymis, an essential organ for sperm maturation and function. Disruption of the development of this structure may result in infertility at adulthood, jus-tifying the need for a better understanding of the mechanisms and mediators driving epididymal mor-phogenesis. Our research group has recently identified the β-defensin SPAG11C (sperm associated an-tigen 11 C), a component of innate immunity, as an androgen-dependent mesenchymal factor that mod-ulates the rat mesonephric duct morphogenesis. We have also detected the expression of other β-defen-sins in this tissue. Members of this protein family can promote their effects via Toll-like receptor (TLR) cellular signaling. Taken together, the present aims of the study were to characterize the expression profile and the androgen regulation of a subset of β-defensins in the rat mesonephric duct, to investigate the functional aspects of the DEFB2 (defensin beta 2) and its potential involvement with TLR4 signaling pathway during the prenatal epididymal development, as well as to validate an experimental approach for the detection of recombinant β-defensin processing and secretion in a cell culture model. Our results revealed that the tested β-defensins presented differential gene expression, androgen regulation and cel-lular distribution profile during prenatal epididymal morphogenesis in rat. The β-defensin Defb4 tran-script, the homologue of human DEFB2 (hDEFB2), was negatively regulated between the embryonic days (e) 17.5 and 20.5, a period when the mesonephric duct morphological differentiation occurs. In ducts (e17.5) cultured ex vivo for 96 h, recombinant hDEFB2 reduced duct elongation and coiling as a result of increased cell death in both epithelial and mesenchymal cells. We have also observed that TLR4-induced signaling pathway is present and functional in the mesonephric duct, and its activation by LPS from E. coli can be negatively regulated by hDEFB2. Furthermore, we have validated a Western blot protocol for the detection of SPAG11C protein forms on cell lysate and supernatant from cultured human cells transiently transfected to express this β-defensin. Collectively, these results reveal the par-ticipation of innate immunity components on the scenario of mesonephric duct development, contrib-uting to increased knowledge of the functional repertoire and role of β-defensins in the epididymis and in events related to male fertility.