Expressão dos membros da subfamília do fator de crescimento fibroblástico 8 (FGF8, FGF17 e FGF18) e dos receptores de fatores de crescimento fibroblástico(FGFRs) durante o desenvolvimento e regressão do corpo do lúteo bovino
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Data
2010-06-30
Autores
Guerra, Diego Marcondes [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A compreensão dos mecanismos moleculares controladores do desenvolvimento, função e regressão do CL bovino é necessária para o aprimoramento da manipulação hormonal ovariana. Fortes evidências sugerem o envolvimento de fatores de crescimento fibroblástico (FGFs) na regulação do crescimento e regressão do CL. “Splicing” alternativo de 4 genes formam sete subtipos de FGFRs com afinidade variável por diferentes FGFs. Os membros da subfamília do FGF8 (FGF8, 17 e 18) ativam eficientemente o FGFR3C e 4 e podem atuar em cooperação nos tecidos que expressão estes receptores. O objetivo deste trabalho foi determinar o padrão de expressão dos FGFRs e dos membros da subfamília do FGF8 no CL bovino (CL). Os CLs foram obtidos de ovários de abatedouro e classificados em 4 estádios de desenvolvimento (estádio/1= corpo hemorrágico, estádio/2= CL em desenvolvimento, estádio/3= CL maduro/início da luteólise funcional e estádio/4= luteólise estrutural). O RNAm foi mensurado por PCR semiquantitativo e a proteína localizada por imunohistoquímica. A expressão do RNAm codificante das isoformas ‘B’ e ‘C’ de FGFR1 e FGFR2 foi detectada no CL bovino por PCR associado à eletroforese e foi acompanhada pela localização da proteína nas pequenas e grandes células luteínicas. A expressão do RNAm do FGFR1C e 2C não variou durante o desenvolvimento luteínico, distintamente a expressão do FGFR1B aumentou no estádio 3. Embora os FGFRs 3B, 3C e 4 tenham sido detectados de forma inconsistente por PCR associado à eletroforese, o RNAm do FGFR3C e FGFR4 foram detectados por PCR em tempo real em todos os estádios do desenvolvimento luteínico. O RNAm do FGF18 foi detectado por PCR em tempo real em todos os estádios do desenvolvimento luteínico e sua abundancia do RNAm do FGF18 foi maior no estádio 3 comparado com os estádios 1, 2 e 4. Em contraste, os RNAm do FGF8 e 17...
The molecular mechanisms controlling the development, function and regression of the bovine corpus luteum are necessary for the improvement of reproductive biotechnologies. Strong evidence suggests the involvement of fibroblast growth factors (FGFs) in the regulation of growth, and regression of the corpus luteum (CL). Alternative splicing of 4 genes give rise to seven subtypes of FGFRs with varying affinity for different FGFs. FGF8 subfamily members (FGF8, 17 and 18) efficiently activate FGFR3C and FGFR4 and may act in cooperation in tissues expressing these receptors. The objective of the present study was to determine the pattern of expression of FGF8 subfamily members and FGFRs in the bovine CL. Bovine CLs were obtained from abattoir ovaries and classed into four stages of development (stage 1= corpus hemorragicum, stage 2= developing CL, stage 3= mature/early functional luteolysis CL, and stage 4= structural luteolysis). Expression of mRNA was measured by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) followed by gel analysis (FGFR1-4) and real time RT-PCR (FGF8 subfamily members, FGFR3C and FGFR4) and proteins were localized by immunohistochemistry. Expression of mRNA encoding ‘B’ and ‘C’ spliced forms of FGFR1 and FGFR2 was readily detected in the bovine CL and was accompanied by isoform non-specific protein localization. FGFR1C and FGFR2C mRNA expression did not vary throughout CL lifespan, whereas FGFR1B was upregulated in the mature CL (stage III). FGFR3B, FGFR3C and FGFR4 expression was inconsistent in the bovine CL as assessed by PCR associated with gel analysis. FGF18, FGFR3C and FGFR4 mRNA was detected by real time PCR in all four developmental stages, and FGF18 mRNA abundance was higher in stage 3 (2.89 0.05; mean ± SEM) compared with stages 1 (0.3 0.27), 2 (0.56 1.27) and 4 (0.99 0.32). The m RNA expression ... (Complete abstract click electronic access below)
The molecular mechanisms controlling the development, function and regression of the bovine corpus luteum are necessary for the improvement of reproductive biotechnologies. Strong evidence suggests the involvement of fibroblast growth factors (FGFs) in the regulation of growth, and regression of the corpus luteum (CL). Alternative splicing of 4 genes give rise to seven subtypes of FGFRs with varying affinity for different FGFs. FGF8 subfamily members (FGF8, 17 and 18) efficiently activate FGFR3C and FGFR4 and may act in cooperation in tissues expressing these receptors. The objective of the present study was to determine the pattern of expression of FGF8 subfamily members and FGFRs in the bovine CL. Bovine CLs were obtained from abattoir ovaries and classed into four stages of development (stage 1= corpus hemorragicum, stage 2= developing CL, stage 3= mature/early functional luteolysis CL, and stage 4= structural luteolysis). Expression of mRNA was measured by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) followed by gel analysis (FGFR1-4) and real time RT-PCR (FGF8 subfamily members, FGFR3C and FGFR4) and proteins were localized by immunohistochemistry. Expression of mRNA encoding ‘B’ and ‘C’ spliced forms of FGFR1 and FGFR2 was readily detected in the bovine CL and was accompanied by isoform non-specific protein localization. FGFR1C and FGFR2C mRNA expression did not vary throughout CL lifespan, whereas FGFR1B was upregulated in the mature CL (stage III). FGFR3B, FGFR3C and FGFR4 expression was inconsistent in the bovine CL as assessed by PCR associated with gel analysis. FGF18, FGFR3C and FGFR4 mRNA was detected by real time PCR in all four developmental stages, and FGF18 mRNA abundance was higher in stage 3 (2.89 0.05; mean ± SEM) compared with stages 1 (0.3 0.27), 2 (0.56 1.27) and 4 (0.99 0.32). The m RNA expression ... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Farmacologia, Bovino - Crescimento, Bovino - Melhoramento genetico, Fibroblast growth factors(FGFs)
Como citar
GUERRA, Diego Marcondes. Expressão dos membros da subfamília do fator de crescimento fibroblástico 8 (FGF8, FGF17 e FGF18) e dos receptores de fatores de crescimento fibroblástico(FGFRs) durante o desenvolvimento e regressão do corpo do lúteo bovino. 2010. 79 f. Dissertação (mestrado) - Universidade Estadual Paulista, Instituto de Biociências de Botucatu, 2010.