title: Proteome Profile and Quantitative Proteomic Analysis of Buffalo (Bubalusbubalis) Follicular Fluid during Follicle Development.
abstract: Follicular fluid (FF) accumulates in the antrum of the ovarian follicle and provides the microenvironment for oocyte development. FF plays an important role in follicle growth and oocyte maturation. The FF provides a unique window to investigate the processes occurring during buffalo follicular development. The observed low quality of buffalo oocytes may arise from the poor follicular microenvironment. Investigating proteins found in buffalo FF (BFF) should provide insight into follicular development processes and provide further understanding of intra-follicular maturation and oocytes quality. Here, a proteomic-based approach was used to analyze the proteome of BFF. SDS-PAGE separation combined with mass spectrometry was used to generate the proteomic dataset. In total, 363 proteins were identified and classified by Gene Ontology terms. The proteins were assigned to 153 pathways, including signaling pathways. To evaluate difference in proteins expressed between BFF with different follicle size (small, <4 mm; and large, >8 mm), a quantitative proteomic analysis based on multi-dimensional liquid chromatography pre-fractionation tandem Orbitrap mass spectrometry identification was performed. Eleven differentially expressed proteins (six downregulated and five upregulated in large BFF) were identified and assigned to a variety of functional processes, including serine protease inhibition, oxidation protection and the complement cascade system. Three differentially expressed proteins, Vimentin, Peroxiredoxin-1 and SERPIND1, were verified by Western blotting, consistent with the quantitative proteomics results. Our datasets offers new information about proteins present in BFF and should facilitate the development of new biomarkers. These differentially expressed proteins illuminate the size-dependent protein changes in follicle microenvironment.
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresource, Guangxi University, Nanning 530004, China. gxfuq@163.com.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Fu
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresource, Guangxi University, Nanning 530004, China. echomide@163.com.
authors.forename: Yulin
authors.initials: Y
authors.lastname: Huang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresource, Guangxi University, Nanning 530004, China. zhqwang@gxu.edu.cn.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Wang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresource, Guangxi University, Nanning 530004, China. gxchenfumei@163.com.
authors.forename: Fumei
authors.initials: F
authors.lastname: Chen
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresource, Guangxi University, Nanning 530004, China. delun.huang@uconn.edu.
authors.forename: Delun
authors.initials: D
authors.lastname: Huang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresource, Guangxi University, Nanning 530004, China. luyangqing@126.com.
authors.forename: Yangqing
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Water Buffalo Institute, Chinese Academy of Agricultural Science, Nanning 530001, China. liangbri@126.com.
authors.forename: Xianwei
authors.initials: X
authors.lastname: Liang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresource, Guangxi University, Nanning 530004, China. mingzhang@gxu.edu.cn.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002020:Buffaloes / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004958:Estradiol / Q000032:analysis; D005260:Female; D015571:Follicular Fluid / Q000378:metabolism; D015844:Heparin Cofactor II / Q000378:metabolism; D006080:Ovarian Follicle / Q000254:growth & development / Q000378:metabolism*; D010455:Peptides / Q000032:analysis; D054464:Peroxiredoxins / Q000378:metabolism; D011374:Progesterone / Q000032:analysis; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D053719:Tandem Mass Spectrometry; D014746:Vimentin / Q000378:metabolism
chemical_list: D010455:Peptides; D020543:Proteome; D014746:Vimentin; D011374:Progesterone; D004958:Estradiol; D015844:Heparin Cofactor II; D054464:Peroxiredoxins
keywords: LC-MS/MS; buffalo; follicular fluid; proteome; quantitative proteomic