Grb10 characterization in bovine cumulus oocyte complexes from different follicle sizes

被引：1
作者：
Antunes da Rosa, Paulo Roberto [1 ]
Bohrer, Rodrigo Camponogara [1 ]
Ludke, Charles Alencar [1 ]
De Cesaro, Matheus Pedroti [1 ]
Pereira, Gabriel Ribas [1 ]
Mondadori, Rafael Gianela [2 ]
Antoniazzi, Alfredo Quites [1 ]
Dias Goncalves, Paulo Bayard [1 ]
机构：
[1] Univ Fed Santa Maria, Hosp Vet Univ, Dept Clin Grandes Anim, Lab Biotecnol & Reprod Anim BioRep, BR-97105900 Santa Maria, RS, Brazil
[2] Univ Fed Pelotas UFPel, Lab Reprod Anim ReproPEL, Pelotas, RS, Brazil
来源：
CIENCIA RURAL | 2015年 / 45卷 / 05期
关键词：
mRNA; protein; maturation; oocyte; Grb10; GROWTH-FACTOR-I; DEVELOPMENTAL COMPETENCE; SIGNALING PATHWAY; VITRO MATURATION; GRANULOSA-CELLS; INSULIN; PROTEIN; DISRUPTION; INHIBITION; ACTIVATION;
D O I：
10.1590/0103-8478cr20140762
中图分类号：
S3 [农学（农艺学）];
学科分类号：
0901 ;
摘要：
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.
引用
收藏
页码：898 / 904
页数：7
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