牛优势卵泡和非优势卵泡颗粒细胞的蛋白质组学特征。

Proteomic characterization of bovine granulosa cells in dominant and subordinate follicles.

机构信息

1College of Life Science, Shanxi Agricultural University, Taigu, 030801 Shanxi China.

2College of Animal Science and Technology, Shanxi Agricultural University, Taigu, 030801 Shanxi China.

出版信息

Hereditas. 2019 Jun 25;156:21. doi: 10.1186/s41065-019-0097-5. eCollection 2019.

DOI:10.1186/s41065-019-0097-5

PMID:31293364

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6593542/

Abstract

BACKGROUND

Characterization of molecular factors regulating ovarian follicular development is critical to understanding its functional mechanism of controlling the estrous cycle, determining oocyte competency, and regulating ovulation. In previous studies, we performed next-gene sequencing to investigate the differentially expressed transcripts of bovine follicular granulosa cells (GCs) at the dominant follicle (DF) and subordinate follicle (SF) stages during the first follicular wave. This study aims to investigate the proteomic characterization of GCs of DF and SF in the bovine estrous cycle.

RESULTS

In total, 3409 proteins were identified from 30,321 peptides obtained from liquid chromatograph-mass spectrometer analysis. Two hundred fifty-nine of these proteins were found to be expressed differently in DF and SF. Out of 259, a total of 26 proteins were upregulated (fold change≥2) and 233 proteins were downregulated (fold change≤0.5) in DF. Gene Ontology (GO) analysis of proteome data revealed the biological process, cellular component and molecular function of expressed proteins in DF and SF, while the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed important signaling pathways associated with follicular development such as the PI3K-Akt, estrogen, and insulin signaling pathways. Immunoblotting results of OGN, ROR2, and HSPB1 confirmed the accuracy of the data. Bioinformatics analysis showed that 13 proteins may be linked to follicular development.

CONCLUSIONS

Findings from this study will provide useful information for exploring follicular development and function.

摘要

背景

阐明调控卵巢卵泡发育的分子因子对于理解其控制发情周期、决定卵母细胞活力和调控排卵的功能机制至关重要。在之前的研究中，我们进行了下一代测序，以研究牛发情周期中第一个卵泡波中优势卵泡（DF）和次级卵泡（SF）阶段颗粒细胞（GC）的差异表达转录本。本研究旨在研究牛发情周期中 DF 和 SF 的 GC 蛋白质组学特征。

结果

从液相色谱-质谱分析获得的 30321 个肽中鉴定出 3409 种蛋白质。其中 259 种在 DF 和 SF 中表达不同。在 259 种中，DF 中有 26 种蛋白上调（倍数变化≥2），233 种蛋白下调（倍数变化≤0.5）。对蛋白质组数据的基因本体（GO）分析显示了 DF 和 SF 中表达蛋白的生物学过程、细胞成分和分子功能，而京都基因与基因组百科全书（KEGG）途径分析则显示了与卵泡发育相关的重要信号通路，如 PI3K-Akt、雌激素和胰岛素信号通路。OGN、ROR2 和 HSPB1 的免疫印迹结果证实了数据的准确性。生物信息学分析表明，13 种蛋白质可能与卵泡发育有关。