• Refine Query
  • Source
  • Publication year
  • to
  • Language
  • 1
  • Tagged with
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Expressão gênica do receptor do hormônio luteinizante (LHR), em células da teca e da granulosa de folículos antrais bovinos /

Nogueira, Marcelo Fábio Gouveia. January 2005 (has links)
Orientador: Ciro Moraes Barros / Resumo: Em células da teca e da granulosa, de folículos bovinos, foram detectados quatro transcritos alternativos do receptor do hormônio luteinizante (LHR). Apenas dois deles são traduzidos em proteínas funcionais com afinidades distintas em relação aos ligantes. Em humanos e símios, a isoforma completa ("full-length") tem afinidade pelo LH e hCG, enquanto que a isoforma que apresenta deleção do exon 10 tem afinidade somente pelo hCG. Além disso, isoformas com deleção do exon 3 foram observadas em ratos, embora nenhum outro estudo tenha investigado essa região do gene bovino. Objetivouse com este trabalho caracterizar o padrão da expressão do gene do LHR nas células da teca e da granulosa de folículos antrais bovinos. Ovários foram coletados em matadouro, os folículos (5-14 mm) foram dissecados e as células da teca e da granulosa separadas para extração de RNA total com Trizol. As concentrações de esteróides no fluido folicular foram determinadas por radioimunoensaio (RIE). A expressão gênica do LHR foi mensurada por RTPCR semiquantitativo com oligonucleotídeos iniciadores ("primers") específicos para amplificar o fragmento entre o final da região extracelular e o final da intracelular (LHRBC; "primers" posicionados nos exons 9 e 11). A ocorrência de transcritos alternativos oriundos do início da região extracelular (LHRA; "primers" posicionados nos exons 2 e 9) foi investigada mediante amplificação por RT-PCR. Como controle interno no PCR, utilizou-se a expressão da GAPDH. Paralelamente, células da granulosa cultivadas in vitro foram tratadas com 1 ou 10 ng de FSH no meio de cultura. Mediante RT-PCR, foi investigada a expressão das isoformas do LHRBC nas células da granulosa cultivadas e tratadas com FSH... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Growth of dominant follicle in the absence of circulating FSH and the events following the LH surge that culminate in ovulation, are dependent on the interaction between LH and its receptor (LHR). Four LHR alternative transcripts were described in theca and granulosa cells from bovine follicles. Only two of them can be translated to functional proteins (receptors coupled with G protein) with different affinities to their ligands. In humans and marmosets, the full-length isoform has affinity to both LH and hCG molecules, whereas the isoform with deletion of only exon 10 has affinity to hCG exclusively. Additionally, isoforms with deletion of exon 3 were observed in rats, although no previous report have investigated this region of the bovine gene. The objective of this study was to characterize the pattern of gene expression of the LHR in theca and granulosa cells from bovine antral follicles. Additionally, LHR expression was determined in cultured granulosa cells under FSH treatment. From ovaries collected in abattoir, antral follicles were dissected (5 to 14mm of diameter), and samples of theca and granulosa cells were obtained to total RNA extraction (Trizol protocol). Steroids concentrations in the follicular fluid were determined by RIA. Gene expression of LHR was measured by semiquantitative RT-PCR with specific primers to amplify part of extracellular region (LHRA; primers annealing on exons 2 and 9) and the fragment from the end of extracellular region, including the transmembrane domain and finishing near the end of intracellular region (LHRBC; primers annealing on exons 9 and 11). As internal control of the PCR, it was used GAPDH expression. Cultured granulosa cells were treated with 0, 1 or 10 ng of FSH (3 replicates each dose). As in vivo and positive control, theca cell sample was utilized to comparison... (Complete abstract, access undermentioned electronic address) / Doutor

Page generated in 0.0289 seconds