Expressão gênica dos fatores de crescimento fibroblástico 7 e 10 (FGF-7 e FGF-10) ao longo do desenvolvimento folicular ovariano bovino
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Data
2006
Autores
Pinto, Maicon Gaissler Lorena [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Estudos recentes têm evidenciado o envolvimento de diferentes FGFs no controle do desenvolvimento folicular ovariano, entre eles o FGF-7. O FGF- 10 apresenta alta similaridade ao FGF-7, expresso em células da teca, e ambos atuam através do mesmo receptor FGFR-2b, expresso em células da granulosa. O FGF-10 modula a atividade celular em uma série de tecidos, mas ainda não foi investigado no ovário. O objetivo principal deste trabalho foi avaliar a participação do FGF-10 no controle do desenvolvimento folicular mediante investigação da expressão do seu RNAm em folículos ovarianos bovinos. A expressão do FGF-7 também foi investigada paralelamente. Além disso, a expressão gênica do FGF-10 e FGF-7 foi investigada numa variedade de tecidos bovinos fetais e adultos. “Pools” de folículos pré-antrais e oócitos, folículos inteiros com diâmetro entre 2 e 4mm, células da teca e da granulosa de folículos maiores que 5mm e amostras de tecidos tiveram seu RNA total extraído. Dos folículos maiores que 5mm, o fluído folicular foi coletado e as concentrações de estradiol e de progesterona foram determinadas por radioimunoensaio. Folículos com progesterona acima de 100ng/mL foram considerados atrésicos e excluídos da análise. Os folículos restantes foram agrupados de acordo com a concentração de estradiol (<5; 5-20; >20-100 e >100ng/mL) e de acordo com o diâmetro (5-7; 8-10 e >10mm). A expressão gênica do FGF-7 e do FGF-10 foi examinada por RT-PCR. Os efeitos da concentração intrafolicular de estradiol e do diâmetro foram investigados por RT-PCR semiquantitativo. A expressão do FGF-10 foi detectada em folículos pré-antrais primordiais, primários e secundários, oócitos e células da teca, mas não em células da granulosa. Adicionalmente, o RNAm do FGF-7 foi pela primeira vez detectado...
Recent studies have evidenced the involvement of different FGF on ovarian follicular development control, among them FGF-7. FGF-10 exhibits high similarity to FGF-7, which is expressed in theca cells, and both act through the same receptor FGFR-2b, expressed in granulosa cells. FGF-10 modulates cellular activities in several tissues, but it has not been investigated in ovary yet. The main objective of this work was to evaluate FGF-10 participation in controlling follicular development, through the investigation of its mRNA expression in bovine ovarian follicles. In parallel, FGF-7 gene expression was also investigated. Furthermore, FGF-10 and FGF-7 gene expression was assessed in a variety of bovine adult and fetal tissues. Pools of preantral follicles and oocytes, whole follicles at 2-4mm in diameter, granulosa and theca cells from follicles larger than 5mm and tissue samples were submitted to total RNA extraction. Follicular fluid from follicles larger than 5 mm was collected and estradiol and progesterone concentrations were measured by RIA. Follicles with progesterone levels above 100ng/mL were considered atretic and excluded from the analysis. Remaining follicles were grouped according to intrafollicular estradiol concentration (<5; 5-20; >20-100; and >100ng/mL) and according to diameter (5-7; 8-10 and >10mm). FGF-7 and FGF-10 gene expression was examined by RT-PCR. Effects of intrafollicular estradiol concentration and diameter were assessed by semiquantitative RT-PCR. FGF-10 gene expression was detected in primordial, primary and secondary preantral follicles, oocytes and theca cells, but not in granulosa cells. In addition, FGF-7 mRNA was detected for the first time in bovine preantral follicles and investigated in oocytes, where it was absent. FGF-10 expression in theca cells decreased as intrafollicular... (Complete abstract click electronic access below)
Recent studies have evidenced the involvement of different FGF on ovarian follicular development control, among them FGF-7. FGF-10 exhibits high similarity to FGF-7, which is expressed in theca cells, and both act through the same receptor FGFR-2b, expressed in granulosa cells. FGF-10 modulates cellular activities in several tissues, but it has not been investigated in ovary yet. The main objective of this work was to evaluate FGF-10 participation in controlling follicular development, through the investigation of its mRNA expression in bovine ovarian follicles. In parallel, FGF-7 gene expression was also investigated. Furthermore, FGF-10 and FGF-7 gene expression was assessed in a variety of bovine adult and fetal tissues. Pools of preantral follicles and oocytes, whole follicles at 2-4mm in diameter, granulosa and theca cells from follicles larger than 5mm and tissue samples were submitted to total RNA extraction. Follicular fluid from follicles larger than 5 mm was collected and estradiol and progesterone concentrations were measured by RIA. Follicles with progesterone levels above 100ng/mL were considered atretic and excluded from the analysis. Remaining follicles were grouped according to intrafollicular estradiol concentration (<5; 5-20; >20-100; and >100ng/mL) and according to diameter (5-7; 8-10 and >10mm). FGF-7 and FGF-10 gene expression was examined by RT-PCR. Effects of intrafollicular estradiol concentration and diameter were assessed by semiquantitative RT-PCR. FGF-10 gene expression was detected in primordial, primary and secondary preantral follicles, oocytes and theca cells, but not in granulosa cells. In addition, FGF-7 mRNA was detected for the first time in bovine preantral follicles and investigated in oocytes, where it was absent. FGF-10 expression in theca cells decreased as intrafollicular... (Complete abstract click electronic access below)
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Reprodução animal, Bovino
Como citar
PINTO, Maicon Gaissler Lorena. Expressão gênica dos fatores de crescimento fibroblástico 7 e 10 (FGF-7 e FGF-10) ao longo do desenvolvimento folicular ovariano bovino. 2006. 83 f. Tese (doutorado) - Universidade Estadual Paulista, Faculdade de Medicina Veterinária e Zootecnia de Botucatu, 2006.