Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes
Autor(a) principal: | |
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Data de Publicação: | 2015 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Ciência Rural |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782015000500898 |
Resumo: | 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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Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes: mRNAproteinmaturationoocyteGrb10.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.Universidade Federal de Santa Maria2015-05-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782015000500898Ciência Rural v.45 n.5 2015reponame:Ciência Ruralinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM10.1590/0103-8478cr20140762info:eu-repo/semantics/openAccessRosa,Paulo Roberto Antunes daBohrer,Rodrigo CamponogaraLudke,Charles AlencarCesaro,Matheus Pedroti DePereira,Gabriel RibasMondadori,Rafael GianelaAntoniazzi,Alfredo QuitesGonçalves,Paulo Bayard Diaseng2015-12-04T00:00:00ZRevista |
dc.title.none.fl_str_mv |
Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes |
title |
Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes |
spellingShingle |
Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes Rosa,Paulo Roberto Antunes da : mRNA protein maturation oocyte Grb10. |
title_short |
Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes |
title_full |
Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes |
title_fullStr |
Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes |
title_full_unstemmed |
Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes |
title_sort |
Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes |
author |
Rosa,Paulo Roberto Antunes da |
author_facet |
Rosa,Paulo Roberto Antunes da Bohrer,Rodrigo Camponogara Ludke,Charles Alencar Cesaro,Matheus Pedroti De Pereira,Gabriel Ribas Mondadori,Rafael Gianela Antoniazzi,Alfredo Quites Gonçalves,Paulo Bayard Dias |
author_role |
author |
author2 |
Bohrer,Rodrigo Camponogara Ludke,Charles Alencar Cesaro,Matheus Pedroti De Pereira,Gabriel Ribas Mondadori,Rafael Gianela Antoniazzi,Alfredo Quites Gonçalves,Paulo Bayard Dias |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
Rosa,Paulo Roberto Antunes da Bohrer,Rodrigo Camponogara Ludke,Charles Alencar Cesaro,Matheus Pedroti De Pereira,Gabriel Ribas Mondadori,Rafael Gianela Antoniazzi,Alfredo Quites Gonçalves,Paulo Bayard Dias |
dc.subject.por.fl_str_mv |
: mRNA protein maturation oocyte Grb10. |
topic |
: mRNA protein maturation oocyte Grb10. |
description |
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. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-05-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782015000500898 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782015000500898 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/0103-8478cr20140762 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria |
dc.source.none.fl_str_mv |
Ciência Rural v.45 n.5 2015 reponame:Ciência Rural instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Ciência Rural |
collection |
Ciência Rural |
repository.name.fl_str_mv |
|
repository.mail.fl_str_mv |
|
_version_ |
1749140547156574208 |