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.

Proteomic profiling of follicle fluids after superstimulation in one‐month‐old lambs

“…The most abundant protein, albumin, as well as other proteins of high relative abundance, including complement C3, apolipoprotein A‐I, serotransferrin and gelsolin, observed here have been reported in studies of other species (Fu et al., ; Zamah, Hassis, Albertolle, & Williams, ). In comparison with other reports exploring various veterinary species as well as human FF (Fu et al., ; Zamah et al., ), it was observed that 119 proteins are conserved among mammalian FF (see Figure and Table ). These unique protein component characteristics of FF likely are a unique microenvironment and affect follicular development and maturation.…”

“…These secretions contain numerous proteins, metabolites, ionic compounds and regulatory factors (Fortune, ). Follicular fluid proteins, such as insulin‐like growth factor‐1 (IGF‐1), tumour necrosis factor alpha (TNFα), interleukin‐6 (IL‐6), vimentin, peroxiredoxin‐1 and SERPIND1, have been shown to directly or indirectly affect the fertility competence of oocytes and their developmental potential (Fu et al., ; Mendoza et al., ; Twigt, Steegers‐Theunissen, Bezstarosti, & Demmers, ). A number of studies have shown that FF inhibits oocyte maturation and embryo development (Wang et al., ; Wu et al., ).…”

Complement C3 and anti-testosterone antibody in the 150 kDa Bali cattle (Bos sondaicus/javanicus) subordinate follicular fluid peptide observed using MALDI-TOF/TOF-MS

“…The presence of Complement C3 has been reported in the human follicular fluid, and it was abundant in younger group as compared to older group (Hashemitabar et al, 2014). Complement C3 has also identified in the mare follicular fluid (Fahiminiya et al, 2010), in the buffalo (Fu et al, 2016); but is limited in the cows. Identification of Complement C3 in Bali cattle follicular fluid is therefore categorized as novel.…”

“…It contains a variety of proteins or peptides that provide the micro environment for follicle development and oocyte maturation (Twigt et al, 2012;Zamah et al, 2015). It has been reported elsewhere, oocyte maturation is related with the presence of gonadotropins and steroids in the FF (Mason et Fu et al (2016) have successfully identified at least eleven differentially expressed proteins from small (<4 mm) and large (>8 mm) buffalo follicular fluid that are involved in serine protease inhibition, complement cascade system and oxidation protection system. From the findings, it was revealed that some proteins or peptides are of common proteins or factors while several are of species-specific.…”

Application of Some Proteome Analysis Techniques in Animal Reproduction

“…Alpha-1-antiproteinase, metalloproteinase inhibitor 2, inter-alphatrypsin inhibitor heavy chain H1, basement membrane-specific heparin sulfate proteoglycan core protein, complement component C8 alpha chain, collagen alpha-2(I) chain, prothrombin, alpha-S1-casein, interleukin-1 receptor accessory protein, and one uncharacterized protein (its gene named LOC781004) were differentially expressed in low-fertile dairy cows compared to controls [13]. Other studies identified 143 proteins in Holstein cows' follicular fluid during follicle development [14] and 363 proteins in buffalos follicular fluid [15], while other studies identified 38 and 463 protein spots in the female dog [16] and the mare [17].…”