Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes

The objective of this study was to investigate the mRNA expression and protein localization of Grb10 gene in bovine cumulus-oocyte complexes (COCs) from different follicle sizes. Firstly, it was investigated the mRNA expression to correlate with maturation rates. COCs from follicles at 1-3, 4-6, 6-8...
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Autor*in:	Paulo Roberto Antunes da Rosa [verfasserIn] Rodrigo Camponogara Bohrer [verfasserIn] Charles Alencar Ludke [verfasserIn] Matheus Pedroti De Cesaro [verfasserIn] Gabriel Ribas Pereira [verfasserIn] Rafael Gianela Mondadori [verfasserIn] Alfredo Quites Antoniazzi [verfasserIn] Paulo Bayard Dias Gonçalves [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch ; Portugiesisch

Erschienen:	2015

Schlagwörter:	O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito. RNAm proteína maturação oócito Grb10.

Übergeordnetes Werk:	In: Ciência Rural - Universidade Federal de Santa Maria, 2004, 45(2015), 5, Seite 898-904
Übergeordnetes Werk:	volume:45 ; year:2015 ; number:5 ; pages:898-904

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1590/0103-8478cr20140762

Katalog-ID:	DOAJ010698116

Internformat


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520			\|a The objective of this study was to investigate the mRNA expression and protein localization of Grb10 gene in bovine cumulus-oocyte complexes (COCs) from different follicle sizes. Firstly, it was investigated the mRNA expression to correlate with maturation rates. COCs from follicles at 1-3, 4-6, 6-8 and >8mm were used to evaluate Grb10 gene expression by qRT-PCR assay and nuclear maturation rates. It was observed that more competent oocytes (from follicles at 6-8 and >8mm; P>0.05), had lower Grb10 mRNA expression levels when compared to the oocytes from follicles at 1-3 and 4-6mm (P>0.05). After it was performed an immunofluorescence analysis in COCs from different follicle sizes (1-3, 4-6, 6-8 and >8mm) to investigate Grb10 protein localization. Samples were incubated with primary antibody: Polyclonal rabbit anti-Grb10 (1:100). Primary antibody was detected using goat anti-rabbit IgG antibody conjugated with Alexa Fluor 488 (1:500). Positive fluorescence signal was detected in all analyzed samples but less evident in COCs from largest follicles. These results characterized Grb10 gene in bovine COC and provide evidences for its involvement during oocyte molecular maturation.
650		4	\|a O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm
650		4	\|a P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito.
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allfields	10.1590/0103-8478cr20140762 doi (DE-627)DOAJ010698116 (DE-599)DOAJ0ff7fe8d3e8244c69eb7655810e6df92 DE-627 ger DE-627 rakwb eng por S1-972 Paulo Roberto Antunes da Rosa verfasserin aut Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes 2015 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier The objective of this study was to investigate the mRNA expression and protein localization of Grb10 gene in bovine cumulus-oocyte complexes (COCs) from different follicle sizes. Firstly, it was investigated the mRNA expression to correlate with maturation rates. COCs from follicles at 1-3, 4-6, 6-8 and >8mm were used to evaluate Grb10 gene expression by qRT-PCR assay and nuclear maturation rates. It was observed that more competent oocytes (from follicles at 6-8 and >8mm; P>0.05), had lower Grb10 mRNA expression levels when compared to the oocytes from follicles at 1-3 and 4-6mm (P>0.05). After it was performed an immunofluorescence analysis in COCs from different follicle sizes (1-3, 4-6, 6-8 and >8mm) to investigate Grb10 protein localization. Samples were incubated with primary antibody: Polyclonal rabbit anti-Grb10 (1:100). Primary antibody was detected using goat anti-rabbit IgG antibody conjugated with Alexa Fluor 488 (1:500). Positive fluorescence signal was detected in all analyzed samples but less evident in COCs from largest follicles. These results characterized Grb10 gene in bovine COC and provide evidences for its involvement during oocyte molecular maturation. O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito. RNAm proteína maturação oócito Grb10. Agriculture S Agriculture (General) Rodrigo Camponogara Bohrer verfasserin aut Charles Alencar Ludke verfasserin aut Matheus Pedroti De Cesaro verfasserin aut Gabriel Ribas Pereira verfasserin aut Rafael Gianela Mondadori verfasserin aut Alfredo Quites Antoniazzi verfasserin aut Paulo Bayard Dias Gonçalves verfasserin aut In Ciência Rural Universidade Federal de Santa Maria, 2004 45(2015), 5, Seite 898-904 (DE-627)320649970 (DE-600)2025834-3 16784596 nnns volume:45 year:2015 number:5 pages:898-904 https://doi.org/10.1590/0103-8478cr20140762 kostenfrei https://doaj.org/article/0ff7fe8d3e8244c69eb7655810e6df92 kostenfrei http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782015000500898&lng=en&tlng=en kostenfrei https://doaj.org/toc/1678-4596 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_252 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 45 2015 5 898-904
spelling	10.1590/0103-8478cr20140762 doi (DE-627)DOAJ010698116 (DE-599)DOAJ0ff7fe8d3e8244c69eb7655810e6df92 DE-627 ger DE-627 rakwb eng por S1-972 Paulo Roberto Antunes da Rosa verfasserin aut Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes 2015 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier The objective of this study was to investigate the mRNA expression and protein localization of Grb10 gene in bovine cumulus-oocyte complexes (COCs) from different follicle sizes. Firstly, it was investigated the mRNA expression to correlate with maturation rates. COCs from follicles at 1-3, 4-6, 6-8 and >8mm were used to evaluate Grb10 gene expression by qRT-PCR assay and nuclear maturation rates. It was observed that more competent oocytes (from follicles at 6-8 and >8mm; P>0.05), had lower Grb10 mRNA expression levels when compared to the oocytes from follicles at 1-3 and 4-6mm (P>0.05). After it was performed an immunofluorescence analysis in COCs from different follicle sizes (1-3, 4-6, 6-8 and >8mm) to investigate Grb10 protein localization. Samples were incubated with primary antibody: Polyclonal rabbit anti-Grb10 (1:100). Primary antibody was detected using goat anti-rabbit IgG antibody conjugated with Alexa Fluor 488 (1:500). Positive fluorescence signal was detected in all analyzed samples but less evident in COCs from largest follicles. These results characterized Grb10 gene in bovine COC and provide evidences for its involvement during oocyte molecular maturation. O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito. RNAm proteína maturação oócito Grb10. Agriculture S Agriculture (General) Rodrigo Camponogara Bohrer verfasserin aut Charles Alencar Ludke verfasserin aut Matheus Pedroti De Cesaro verfasserin aut Gabriel Ribas Pereira verfasserin aut Rafael Gianela Mondadori verfasserin aut Alfredo Quites Antoniazzi verfasserin aut Paulo Bayard Dias Gonçalves verfasserin aut In Ciência Rural Universidade Federal de Santa Maria, 2004 45(2015), 5, Seite 898-904 (DE-627)320649970 (DE-600)2025834-3 16784596 nnns volume:45 year:2015 number:5 pages:898-904 https://doi.org/10.1590/0103-8478cr20140762 kostenfrei https://doaj.org/article/0ff7fe8d3e8244c69eb7655810e6df92 kostenfrei http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782015000500898&lng=en&tlng=en kostenfrei https://doaj.org/toc/1678-4596 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_252 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 45 2015 5 898-904
allfields_unstemmed	10.1590/0103-8478cr20140762 doi (DE-627)DOAJ010698116 (DE-599)DOAJ0ff7fe8d3e8244c69eb7655810e6df92 DE-627 ger DE-627 rakwb eng por S1-972 Paulo Roberto Antunes da Rosa verfasserin aut Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes 2015 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier The objective of this study was to investigate the mRNA expression and protein localization of Grb10 gene in bovine cumulus-oocyte complexes (COCs) from different follicle sizes. Firstly, it was investigated the mRNA expression to correlate with maturation rates. COCs from follicles at 1-3, 4-6, 6-8 and >8mm were used to evaluate Grb10 gene expression by qRT-PCR assay and nuclear maturation rates. It was observed that more competent oocytes (from follicles at 6-8 and >8mm; P>0.05), had lower Grb10 mRNA expression levels when compared to the oocytes from follicles at 1-3 and 4-6mm (P>0.05). After it was performed an immunofluorescence analysis in COCs from different follicle sizes (1-3, 4-6, 6-8 and >8mm) to investigate Grb10 protein localization. Samples were incubated with primary antibody: Polyclonal rabbit anti-Grb10 (1:100). Primary antibody was detected using goat anti-rabbit IgG antibody conjugated with Alexa Fluor 488 (1:500). Positive fluorescence signal was detected in all analyzed samples but less evident in COCs from largest follicles. These results characterized Grb10 gene in bovine COC and provide evidences for its involvement during oocyte molecular maturation. O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito. RNAm proteína maturação oócito Grb10. Agriculture S Agriculture (General) Rodrigo Camponogara Bohrer verfasserin aut Charles Alencar Ludke verfasserin aut Matheus Pedroti De Cesaro verfasserin aut Gabriel Ribas Pereira verfasserin aut Rafael Gianela Mondadori verfasserin aut Alfredo Quites Antoniazzi verfasserin aut Paulo Bayard Dias Gonçalves verfasserin aut In Ciência Rural Universidade Federal de Santa Maria, 2004 45(2015), 5, Seite 898-904 (DE-627)320649970 (DE-600)2025834-3 16784596 nnns volume:45 year:2015 number:5 pages:898-904 https://doi.org/10.1590/0103-8478cr20140762 kostenfrei https://doaj.org/article/0ff7fe8d3e8244c69eb7655810e6df92 kostenfrei http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782015000500898&lng=en&tlng=en kostenfrei https://doaj.org/toc/1678-4596 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_252 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 45 2015 5 898-904
allfieldsGer	10.1590/0103-8478cr20140762 doi (DE-627)DOAJ010698116 (DE-599)DOAJ0ff7fe8d3e8244c69eb7655810e6df92 DE-627 ger DE-627 rakwb eng por S1-972 Paulo Roberto Antunes da Rosa verfasserin aut Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes 2015 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier The objective of this study was to investigate the mRNA expression and protein localization of Grb10 gene in bovine cumulus-oocyte complexes (COCs) from different follicle sizes. Firstly, it was investigated the mRNA expression to correlate with maturation rates. COCs from follicles at 1-3, 4-6, 6-8 and >8mm were used to evaluate Grb10 gene expression by qRT-PCR assay and nuclear maturation rates. It was observed that more competent oocytes (from follicles at 6-8 and >8mm; P>0.05), had lower Grb10 mRNA expression levels when compared to the oocytes from follicles at 1-3 and 4-6mm (P>0.05). After it was performed an immunofluorescence analysis in COCs from different follicle sizes (1-3, 4-6, 6-8 and >8mm) to investigate Grb10 protein localization. Samples were incubated with primary antibody: Polyclonal rabbit anti-Grb10 (1:100). Primary antibody was detected using goat anti-rabbit IgG antibody conjugated with Alexa Fluor 488 (1:500). Positive fluorescence signal was detected in all analyzed samples but less evident in COCs from largest follicles. These results characterized Grb10 gene in bovine COC and provide evidences for its involvement during oocyte molecular maturation. O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito. RNAm proteína maturação oócito Grb10. Agriculture S Agriculture (General) Rodrigo Camponogara Bohrer verfasserin aut Charles Alencar Ludke verfasserin aut Matheus Pedroti De Cesaro verfasserin aut Gabriel Ribas Pereira verfasserin aut Rafael Gianela Mondadori verfasserin aut Alfredo Quites Antoniazzi verfasserin aut Paulo Bayard Dias Gonçalves verfasserin aut In Ciência Rural Universidade Federal de Santa Maria, 2004 45(2015), 5, Seite 898-904 (DE-627)320649970 (DE-600)2025834-3 16784596 nnns volume:45 year:2015 number:5 pages:898-904 https://doi.org/10.1590/0103-8478cr20140762 kostenfrei https://doaj.org/article/0ff7fe8d3e8244c69eb7655810e6df92 kostenfrei http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782015000500898&lng=en&tlng=en kostenfrei https://doaj.org/toc/1678-4596 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_252 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 45 2015 5 898-904
allfieldsSound	10.1590/0103-8478cr20140762 doi (DE-627)DOAJ010698116 (DE-599)DOAJ0ff7fe8d3e8244c69eb7655810e6df92 DE-627 ger DE-627 rakwb eng por S1-972 Paulo Roberto Antunes da Rosa verfasserin aut Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes 2015 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier The objective of this study was to investigate the mRNA expression and protein localization of Grb10 gene in bovine cumulus-oocyte complexes (COCs) from different follicle sizes. Firstly, it was investigated the mRNA expression to correlate with maturation rates. COCs from follicles at 1-3, 4-6, 6-8 and >8mm were used to evaluate Grb10 gene expression by qRT-PCR assay and nuclear maturation rates. It was observed that more competent oocytes (from follicles at 6-8 and >8mm; P>0.05), had lower Grb10 mRNA expression levels when compared to the oocytes from follicles at 1-3 and 4-6mm (P>0.05). After it was performed an immunofluorescence analysis in COCs from different follicle sizes (1-3, 4-6, 6-8 and >8mm) to investigate Grb10 protein localization. Samples were incubated with primary antibody: Polyclonal rabbit anti-Grb10 (1:100). Primary antibody was detected using goat anti-rabbit IgG antibody conjugated with Alexa Fluor 488 (1:500). Positive fluorescence signal was detected in all analyzed samples but less evident in COCs from largest follicles. These results characterized Grb10 gene in bovine COC and provide evidences for its involvement during oocyte molecular maturation. O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito. RNAm proteína maturação oócito Grb10. Agriculture S Agriculture (General) Rodrigo Camponogara Bohrer verfasserin aut Charles Alencar Ludke verfasserin aut Matheus Pedroti De Cesaro verfasserin aut Gabriel Ribas Pereira verfasserin aut Rafael Gianela Mondadori verfasserin aut Alfredo Quites Antoniazzi verfasserin aut Paulo Bayard Dias Gonçalves verfasserin aut In Ciência Rural Universidade Federal de Santa Maria, 2004 45(2015), 5, Seite 898-904 (DE-627)320649970 (DE-600)2025834-3 16784596 nnns volume:45 year:2015 number:5 pages:898-904 https://doi.org/10.1590/0103-8478cr20140762 kostenfrei https://doaj.org/article/0ff7fe8d3e8244c69eb7655810e6df92 kostenfrei http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782015000500898&lng=en&tlng=en kostenfrei https://doaj.org/toc/1678-4596 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_252 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 45 2015 5 898-904
language	English Portuguese
source	In Ciência Rural 45(2015), 5, Seite 898-904 volume:45 year:2015 number:5 pages:898-904
sourceStr	In Ciência Rural 45(2015), 5, Seite 898-904 volume:45 year:2015 number:5 pages:898-904
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito. RNAm proteína maturação oócito Grb10. Agriculture S Agriculture (General)
isfreeaccess_bool	true
container_title	Ciência Rural
authorswithroles_txt_mv	Paulo Roberto Antunes da Rosa @@aut@@ Rodrigo Camponogara Bohrer @@aut@@ Charles Alencar Ludke @@aut@@ Matheus Pedroti De Cesaro @@aut@@ Gabriel Ribas Pereira @@aut@@ Rafael Gianela Mondadori @@aut@@ Alfredo Quites Antoniazzi @@aut@@ Paulo Bayard Dias Gonçalves @@aut@@
publishDateDaySort_date	2015-01-01T00:00:00Z
hierarchy_top_id	320649970
id	DOAJ010698116
language_de	englisch portugiesisch
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Firstly, it was investigated the mRNA expression to correlate with maturation rates. COCs from follicles at 1-3, 4-6, 6-8 and &gt;8mm were used to evaluate Grb10 gene expression by qRT-PCR assay and nuclear maturation rates. It was observed that more competent oocytes (from follicles at 6-8 and &gt;8mm; P&gt;0.05), had lower Grb10 mRNA expression levels when compared to the oocytes from follicles at 1-3 and 4-6mm (P&gt;0.05). After it was performed an immunofluorescence analysis in COCs from different follicle sizes (1-3, 4-6, 6-8 and &gt;8mm) to investigate Grb10 protein localization. Samples were incubated with primary antibody: Polyclonal rabbit anti-Grb10 (1:100). Primary antibody was detected using goat anti-rabbit IgG antibody conjugated with Alexa Fluor 488 (1:500). Positive fluorescence signal was detected in all analyzed samples but less evident in COCs from largest follicles. 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callnumber-first	S - Agriculture
author	Paulo Roberto Antunes da Rosa
spellingShingle	Paulo Roberto Antunes da Rosa misc S1-972 misc O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm misc P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito. misc RNAm misc proteína misc maturação misc oócito misc Grb10. misc Agriculture misc S misc Agriculture (General) Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes
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topic_title	S1-972 Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito RNAm proteína maturação oócito Grb10
topic	misc S1-972 misc O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm misc P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito. misc RNAm misc proteína misc maturação misc oócito misc Grb10. misc Agriculture misc S misc Agriculture (General)
topic_unstemmed	misc S1-972 misc O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm misc P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito. misc RNAm misc proteína misc maturação misc oócito misc Grb10. misc Agriculture misc S misc Agriculture (General)
topic_browse	misc S1-972 misc O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. Foi observado que os oócitos mais competentes para a progressão meiótica (oriundos de folículos com diâmetro de 6-8 e <8mm misc P<0.05) tiveram menor expressão de RNAm para Grb10, quando comparados aos CCOs oriundos de folículos com 1-3 e 4-6mm (P<0.05). Posteriormente, foi realizada a técnica de imunofluorescência em CCOs oriundos de folículos de diferentes tamanhos (1-3, 4-6, 6-8 e <8mm) para investigar a localização da proteína Grb10. As amostras foram incubadas com o anticorpo primário: anti-Grb10 policlonal, produzido em coelho (1:100). O anticorpo primário foi detectado utilizando um anticorpo secundário, IgG anti-coelho, produzido em cabra, conjugado com Alexa Fluor 488 (1:500). A fluorescência foi detectada em todas as amostras analisadas, porém, menos evidente nos CCOs oriundos dos folículos maiores. Os resultados apresentados neste estudo caracterizam o gene Grb10 em CCOs de bovinos e fornecem evidências do seu envolvimento na maturação molecular do oócito. misc RNAm misc proteína misc maturação misc oócito misc Grb10. misc Agriculture misc S misc Agriculture (General)
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title	Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes
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title_full	Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes
author_sort	Paulo Roberto Antunes da Rosa
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author_browse	Paulo Roberto Antunes da Rosa Rodrigo Camponogara Bohrer Charles Alencar Ludke Matheus Pedroti De Cesaro Gabriel Ribas Pereira Rafael Gianela Mondadori Alfredo Quites Antoniazzi Paulo Bayard Dias Gonçalves
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title_sort	grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes
callnumber	S1-972
title_auth	Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes
abstract	The objective of this study was to investigate the mRNA expression and protein localization of Grb10 gene in bovine cumulus-oocyte complexes (COCs) from different follicle sizes. Firstly, it was investigated the mRNA expression to correlate with maturation rates. COCs from follicles at 1-3, 4-6, 6-8 and >8mm were used to evaluate Grb10 gene expression by qRT-PCR assay and nuclear maturation rates. It was observed that more competent oocytes (from follicles at 6-8 and >8mm; P>0.05), had lower Grb10 mRNA expression levels when compared to the oocytes from follicles at 1-3 and 4-6mm (P>0.05). After it was performed an immunofluorescence analysis in COCs from different follicle sizes (1-3, 4-6, 6-8 and >8mm) to investigate Grb10 protein localization. Samples were incubated with primary antibody: Polyclonal rabbit anti-Grb10 (1:100). Primary antibody was detected using goat anti-rabbit IgG antibody conjugated with Alexa Fluor 488 (1:500). Positive fluorescence signal was detected in all analyzed samples but less evident in COCs from largest follicles. These results characterized Grb10 gene in bovine COC and provide evidences for its involvement during oocyte molecular maturation.
abstractGer	The objective of this study was to investigate the mRNA expression and protein localization of Grb10 gene in bovine cumulus-oocyte complexes (COCs) from different follicle sizes. Firstly, it was investigated the mRNA expression to correlate with maturation rates. COCs from follicles at 1-3, 4-6, 6-8 and >8mm were used to evaluate Grb10 gene expression by qRT-PCR assay and nuclear maturation rates. It was observed that more competent oocytes (from follicles at 6-8 and >8mm; P>0.05), had lower Grb10 mRNA expression levels when compared to the oocytes from follicles at 1-3 and 4-6mm (P>0.05). After it was performed an immunofluorescence analysis in COCs from different follicle sizes (1-3, 4-6, 6-8 and >8mm) to investigate Grb10 protein localization. Samples were incubated with primary antibody: Polyclonal rabbit anti-Grb10 (1:100). Primary antibody was detected using goat anti-rabbit IgG antibody conjugated with Alexa Fluor 488 (1:500). Positive fluorescence signal was detected in all analyzed samples but less evident in COCs from largest follicles. These results characterized Grb10 gene in bovine COC and provide evidences for its involvement during oocyte molecular maturation.
abstract_unstemmed	The objective of this study was to investigate the mRNA expression and protein localization of Grb10 gene in bovine cumulus-oocyte complexes (COCs) from different follicle sizes. Firstly, it was investigated the mRNA expression to correlate with maturation rates. COCs from follicles at 1-3, 4-6, 6-8 and >8mm were used to evaluate Grb10 gene expression by qRT-PCR assay and nuclear maturation rates. It was observed that more competent oocytes (from follicles at 6-8 and >8mm; P>0.05), had lower Grb10 mRNA expression levels when compared to the oocytes from follicles at 1-3 and 4-6mm (P>0.05). After it was performed an immunofluorescence analysis in COCs from different follicle sizes (1-3, 4-6, 6-8 and >8mm) to investigate Grb10 protein localization. Samples were incubated with primary antibody: Polyclonal rabbit anti-Grb10 (1:100). Primary antibody was detected using goat anti-rabbit IgG antibody conjugated with Alexa Fluor 488 (1:500). Positive fluorescence signal was detected in all analyzed samples but less evident in COCs from largest follicles. These results characterized Grb10 gene in bovine COC and provide evidences for its involvement during oocyte molecular maturation.
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container_issue	5
title_short	Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes
url	https://doi.org/10.1590/0103-8478cr20140762 https://doaj.org/article/0ff7fe8d3e8244c69eb7655810e6df92 http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782015000500898&lng=en&tlng=en https://doaj.org/toc/1678-4596
remote_bool	true
author2	Rodrigo Camponogara Bohrer Charles Alencar Ludke Matheus Pedroti De Cesaro Gabriel Ribas Pereira Rafael Gianela Mondadori Alfredo Quites Antoniazzi Paulo Bayard Dias Gonçalves
author2Str	Rodrigo Camponogara Bohrer Charles Alencar Ludke Matheus Pedroti De Cesaro Gabriel Ribas Pereira Rafael Gianela Mondadori Alfredo Quites Antoniazzi Paulo Bayard Dias Gonçalves
ppnlink	320649970
callnumber-subject	S - General Agriculture
mediatype_str_mv	c
isOA_txt	true
hochschulschrift_bool	false
doi_str	10.1590/0103-8478cr20140762
callnumber-a	S1-972
up_date	2024-07-03T16:15:49.258Z
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Firstly, it was investigated the mRNA expression to correlate with maturation rates. COCs from follicles at 1-3, 4-6, 6-8 and &gt;8mm were used to evaluate Grb10 gene expression by qRT-PCR assay and nuclear maturation rates. It was observed that more competent oocytes (from follicles at 6-8 and &gt;8mm; P&gt;0.05), had lower Grb10 mRNA expression levels when compared to the oocytes from follicles at 1-3 and 4-6mm (P&gt;0.05). After it was performed an immunofluorescence analysis in COCs from different follicle sizes (1-3, 4-6, 6-8 and &gt;8mm) to investigate Grb10 protein localization. Samples were incubated with primary antibody: Polyclonal rabbit anti-Grb10 (1:100). Primary antibody was detected using goat anti-rabbit IgG antibody conjugated with Alexa Fluor 488 (1:500). Positive fluorescence signal was detected in all analyzed samples but less evident in COCs from largest follicles. These results characterized Grb10 gene in bovine COC and provide evidences for its involvement during oocyte molecular maturation.</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">O objetivo deste estudo foi investigar a expressão de RNAm e a localização da proteína Grb10 em complexos cumulus-oócito (CCO), oriundos de folículos bovinos em diferentes fases de desenvolvimento. Primeiramente, foi investigada a expressão de RNAm e relacionada com as taxas de maturação. CCOs oriundos de folículos com diâmetro de 1-3, 4-6, 6-8 e <8mm foram utilizados para avaliar a expressão de RNAm por PCR em tempo real e para analisar os estádios de maturação nuclear. 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Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes

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