Proteômica do fluído folicular de búfalas
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Data
2020-07-05
Autores
Marques, Nayara Fernanda Silva
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O objetivo do presente estudo foi identificar, quantificar e descrever o perfil proteico do liquido folicular e morfometria dos foliculos, de ovários de búfalas mestiças das raças Mediterrânea e Murrah.O fluido folicular (FF) é um meio essencial para o crescimento e a maturação das células ovarianas. A análise dos seus componentes é fundamental para o entendimento da dinâmica folicular da espécie bubalina. O objetivo deste estudo foi traçar o perfil proteico do FF. Foram utilizados 112 ovários provenientes de um abatedouro, a extração do FF foi realizada manualmente com o uso de seringas e agulhas por meio de punção aspirativa nos folículos visíveis macroscopicamente, os folículos aspirados foram separados em seis grupos, < 5 mm com corpo lúteo (CL); < 5 sem CL; 5 – 8 com CL; 5 – 8 sem CL; > 8 com CL; > 8 sem CL. A concentração de proteína total foi determinada pelo método de Bradford. Baseado na concentração de proteina total uma alíquota contendo 50 µg foi aplicada em gel de poliacrilamida 12%, a corrida foi interrompida quando as amostras atingiram o gel de separação, a seguir foi realizada a disgestão em gel para a análise da espectrometria de massa em Nano-LC-ESI-Q-TOF mass spectrometry. A analise estatística foi realizada utilizando o programa MetaboAnalyst 4.0 e de acordo com os dados obtidos podemos observar no dendrograma e na analise de componente principal (PCA), no Vip Scores e no Heat Map que a concetração de proteínas foi constante em todos os grupos independente do tamanho dos folículos. As proteínas mais relevantes foram, Mucin-19, Haptoglobin, Histone H2A type 1, Histone H2B, Lysosomal alpha-glucosidase, Glutathione S-transferase A1, e a Serine protease inhibitor clade E member 2. Podemos relacionar estas proteínas que estão em abundancia no liquido folicular como sendo promotoras do desenvolvimento folicular, e do processo de maturação oocitária.
The objective of the present study was to identify, quantify and describe the protein profile of the follicular fluid and morphometry of the follicles, of mixedrace buffalo ovaries of the Mediterranean and Murrah breeds. The follicular fluid (FF) is an essential medium for the growth and maturation of the ovarian cells. The analysis of its components is fundamental for understanding the follicular dynamics of the buffalo species. The aim of this study was to trace the protein profile of FF. 112 ovaries from a slaughterhouse were used, FF extraction was performed manually using syringes and needles through aspiration puncture in the macroscopically visible follicles, the aspirated follicles were separated into six groups, <5 mm with corpus luteum (CL ); <5 without CL; 5 - 8 with CL; 5 - 8 without CL; > 8 with CL; > 8 without CL. The total protein concentration was determined by the Bradford method. Based on the total protein concentration, an aliquot containing 50 µg was applied on 12% polyacrylamide gel, the run was interrupted when the samples reached the separation gel, then the gel was analyzed for analysis of the Nano mass spectrometry. -LC-ESI-Q-TOF mass spectrometry. The statistical analysis was performed using the MetaboAnalyst 4.0 program and according to the data obtained, we can see in the dendrogram and in the main component analysis (PCA), in the Vip Scores and in the Heat Map that the concentration of proteins was constant in all groups regardless the size of the follicles. The most relevant proteins were, Mucin-19, Haptoglobin, Histone H2A type 1, Histone H2B, Lysosomal alpha-glucosidase, Glutathione Stransferase A1, e a Serine protease inhibitor clade E member 2. We can relate these proteins that are in abundance in the follicular fluid as being promoters of follicular development, and the oocyte maturation process.
The objective of the present study was to identify, quantify and describe the protein profile of the follicular fluid and morphometry of the follicles, of mixedrace buffalo ovaries of the Mediterranean and Murrah breeds. The follicular fluid (FF) is an essential medium for the growth and maturation of the ovarian cells. The analysis of its components is fundamental for understanding the follicular dynamics of the buffalo species. The aim of this study was to trace the protein profile of FF. 112 ovaries from a slaughterhouse were used, FF extraction was performed manually using syringes and needles through aspiration puncture in the macroscopically visible follicles, the aspirated follicles were separated into six groups, <5 mm with corpus luteum (CL ); <5 without CL; 5 - 8 with CL; 5 - 8 without CL; > 8 with CL; > 8 without CL. The total protein concentration was determined by the Bradford method. Based on the total protein concentration, an aliquot containing 50 µg was applied on 12% polyacrylamide gel, the run was interrupted when the samples reached the separation gel, then the gel was analyzed for analysis of the Nano mass spectrometry. -LC-ESI-Q-TOF mass spectrometry. The statistical analysis was performed using the MetaboAnalyst 4.0 program and according to the data obtained, we can see in the dendrogram and in the main component analysis (PCA), in the Vip Scores and in the Heat Map that the concentration of proteins was constant in all groups regardless the size of the follicles. The most relevant proteins were, Mucin-19, Haptoglobin, Histone H2A type 1, Histone H2B, Lysosomal alpha-glucosidase, Glutathione Stransferase A1, e a Serine protease inhibitor clade E member 2. We can relate these proteins that are in abundance in the follicular fluid as being promoters of follicular development, and the oocyte maturation process.
Descrição
Palavras-chave
Líquido folicular, Ovário, Folículo, Proteômica, Follicular fluid, Ovary, Follicle, Proteomics