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T. Zakar
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F. J. Teixeira
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J. J. Hirst
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F. Guo
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E. A. MacLeod
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D. M. Olson
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Since glucocorticoids decrease and protein kinase C (PKC) activators increase amniotic PGE2 production, the possibility that they regulate the activity of prostaglandin endoperoxide H synthase (PGHS), the rate-limiting enzyme of prostaglandin synthesis from arachidonate, was investigated. Glucocorticoids inhibited the production of PGE2 from exogenous arachidonate specifically and in a concentration dependent fashion. Furthermore, cortisol decreased PGHS activity and the amount of PGHS protein in amnion microsomes, and reduced the rate of recovery of PGHS after acetylsalicylic acid (ASA) pretreatment. Actinomycin D blocked the inhibition of PGHS recovery by cortisol, but did not suppress the spontaneous recovery of the enzyme, indicating that the glucocorticoid induced a post-transcriptional inhibitor of PGHS synthesis. PKC-activating phorbol esters, such as 12-tetradecanoyl phorbol 13-acetate (TPA) increased the synthesis of PGE2 from exogenous arachidonate, also in a specific and concentration dependent manner. PGHS recovery after ASA treatment was enhanced by TPA. PGHS activity and protein concentrations were increased by phorbol ester treatment; however, this was apparent only in tissues in which the concentrations of PGHS were initially low. These results show that the synthesis of PGHS is positively and negatively regulated in the human amnion by PKC and glucocorticoids, respectively, and suggest that effectors using these pathways may regulate the enzyme in vivo.

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André F A Figueiredo Department of Morphology, Institute of Biological Sciences, Laboratory of Cellular Biology, Federal University of Minas Gerais - UFMG, Belo Horizonte, Minas Gerais, Brazil

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Natália Teixeira Wnuk Department of Morphology, Institute of Biological Sciences, Laboratory of Cellular Biology, Federal University of Minas Gerais - UFMG, Belo Horizonte, Minas Gerais, Brazil

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Amanda O Tavares Department of Morphology, Institute of Biological Sciences, Laboratory of Cellular Biology, Federal University of Minas Gerais - UFMG, Belo Horizonte, Minas Gerais, Brazil

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José Rafael Miranda Department of Veterinary Medicine, Federal University of Lavras, Lavras, Minas Gerais - UFLA, Brazil

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Rex A Hess Department of Comparative Biosciences, University of Illinois, Urbana Champaign, Illinois, USA

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Luiz Renato de França Department of Morphology, Institute of Biological Sciences, Laboratory of Cellular Biology, Federal University of Minas Gerais - UFMG, Belo Horizonte, Minas Gerais, Brazil

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Guilherme M J Costa Department of Morphology, Institute of Biological Sciences, Laboratory of Cellular Biology, Federal University of Minas Gerais - UFMG, Belo Horizonte, Minas Gerais, Brazil

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The number of Sertoli cells (SCs) ultimately determines the upper limit of sperm production in the testis. Previous studies have shown that thyroid hormones (TH) receptors are abundantly expressed in developing SCs; therefore, it was highly significant to discover that transient neonatal hypothyroidism induced by the goitrogen 6-n-propyl-2-thiouracil (PTU) can extend SCs proliferation beyond the first 2 weeks postnatal and increase testis weight and sperm production. Further studies concluded that treatment must begin before day 8 post birth in rats. Recent studies, however, showed that SCs present in the transition region at the rete testis exhibit a more immature phenotype and have prolonged mitotic activity, which led to the hypothesis that SCs in this region will retain the capacity to respond to PTU treatment over a longer period of time. In the present study, male Wistar rats were treated with PTU from days 21 to 40 and were evaluated at 40 and 160 days of age. Similar to neonatal rat SCs, it was demonstrated that prepubertal SCs in the transition region have a high mitotic activity and are highly sensitive to TH levels. This delayed, transient hypothyroidism resulted in significantly increased testis weight, SCs number and daily sperm production. The results demonstrate for the first time that Sertoli cells showing plasticity in the transition region can be stimulated to increase proliferation and contribute to a late stage surge in testis weight and sperm output.

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J Buratini Jr Departamento de Fisiologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Farmacologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil and Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada

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A B Teixeira Departamento de Fisiologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Farmacologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil and Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada

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I B Costa Departamento de Fisiologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Farmacologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil and Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada

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V F Glapinski Departamento de Fisiologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Farmacologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil and Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada

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M G L Pinto Departamento de Fisiologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Farmacologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil and Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada

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I C Giometti Departamento de Fisiologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Farmacologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil and Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada

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C M Barros Departamento de Fisiologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Farmacologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil and Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada

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M Cao Departamento de Fisiologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Farmacologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil and Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada

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E S Nicola Departamento de Fisiologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Farmacologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil and Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada

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C A Price Departamento de Fisiologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil, Departamento de Farmacologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil and Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada

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Paracrine cell signaling is believed to be important for ovarian follicle development, and a role for some members of the fibroblast growth factor (FGF) family has been suggested. In the present study, we tested the hypothesis that FGF-8 and its cognate receptors (FGFR3c and FGFR4) are expressed in bovine antral follicles. RT-PCR was used to analyze bovine Fgf8, Fgfr3c and Fgfr4 mRNA levels in oocytes, and granulosa and theca cells. Fgf8 expression was detected in oocytes and in granulosa and theca cells; this expression pattern differs from that reported in rodents. Granulosa and theca cells, but not oocytes, expressed Fgfr3c, and expression in granulosa cells increased significantly with follicle estradiol content, a major indicator of follicle health. Fgfr4 expression was restricted to theca cells in the follicle, and decreased significantly with increasing follicle size. To investigate the potential regulation of Fgfr3c expression in the bovine granulosa, cells were cultured in serum-free medium with FSH or IGF-I; gene expression was upregulated by FSH but not by IGF-I. The FSH-responsive and developmentally regulated patterns of Fgfr3c mRNA expression suggest that this receptor is a potential mediator of paracrine signaling to granulosa cells during antral follicle growth in cattle.

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