Dataset Information

Seasonal variation in equine follicular fluid proteome.

ABSTRACT: BACKGROUND:Proteomic studies of follicular fluid (FF) exist for several species, including the horse; however, the seasonal influence on FF proteome has not been explored in livestock. The application of high-throughput proteomics of FF in horse has the potential to identify seasonal variations of proteins involved in follicle and oocyte growth. METHODS:This study (i) profiles the proteomes of equine FF collected from dominant growing follicles during the spring anovulatory season (SAN), and spring (SOV), summer (SUM), and fall (FOV) ovulatory seasons; and (ii) identifies season-dependent regulatory networks and associated key proteins. RESULTS:Regardless of season, a total of 90 proteins were identified in FF, corresponding to 63, 72, 69, and 78 proteins detected in the SAN, SOV, SUM, and FOV seasons, respectively. Fifty-two proteins were common to all seasons, a total of 13 were unique to either season, and 25 were shared between two seasons or more. Protein-to-protein interaction (PPI) analysis indicated the likely critical roles of plasminogen in the SAN season, the prothrombin/plasminogen combination in SUM, and plasminogen/complement C3 in both SOV and FOV seasons. The apolipoprotein A1 appeared crucial in all seasons. The present findings show that FF proteome of SUM differs from other seasons, with FF having high fluidity (low viscosity). CONCLUSIONS:The balance between the FF contents in prothrombin, plasminogen, and coagulation factor XII proteins favoring FF fluidity may be crucial at the peak of the ovulatory season (SUM) and may explain the reported lower incidence of hemorrhagic anovulatory follicles during the SUM season.

SUBMITTER: Dutra GA

PROVIDER: S-EPMC6404268 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature

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Seasonal variation in equine follicular fluid proteome.

Dutra G A GA Ishak G M GM Pechanova O O Pechan T T Peterson D G DG Jacob J C F JCF Willard S T ST Ryan P L PL Gastal E L EL Feugang J M JM

Reproductive biology and endocrinology : RB&E 20190306 1

<h4>Background</h4>Proteomic studies of follicular fluid (FF) exist for several species, including the horse; however, the seasonal influence on FF proteome has not been explored in livestock. The application of high-throughput proteomics of FF in horse has the potential to identify seasonal variations of proteins involved in follicle and oocyte growth.<h4>Methods</h4>This study (i) profiles the proteomes of equine FF collected from dominant growing follicles during the spring anovulatory season ...[more]

PMID: 30841911

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Project description:Ovarian follicular fluid (FF) comprises a dynamic milieu whose composition alters under seasonal variation, influencing follicle development and oocyte developmental competence. Various cell subtypes (e.g. granulosa, theca, and cumulus) produce cell-secreted nanoparticles, coined extracellular vesicles (EVs), which contain bioactive regulatory molecules such as microRNAs (miRNAs) known to mediate intrafollicular communication through various forms of signaling and information transfer. Markedly, the hallmark feature of EVs transferring cellular knowledge propagates their appealing nature as direct indicators of cellular status, albeit homeostasis, dysfunction, or disease. Photoperiod alterations influencing the regulatory content of FF-EVs as downstream modifiers remain largely unknown, particularly in seasonal breeding animals. Here, we aimed to unpack the impacts of seasonal variation on the miRNA expression profiles of FF-EVs in the mare. Equine pre-ovulatory follicles were monitored and aspirated using ultrasound-guided transvaginal techniques to obtain follicular fluids during non-breeding (spring anovulatory, SAN) and breeding seasons (spring ovulatory, SOV; summer, SUM; and fall ovulatory, FOV). 97 miRNAs were found to be differentially expressed between groups. Pairwise comparisons revealed specific clusters, including six miRNAs involved in the spring transition (miR-149-200b-206-221-328-615) and many others dysregulated during that of the summer (miR-143-192451- 302b-100, and let 7c), the peak time point for breeding. Bioinformatic analyses led to unveiling significant enrichments in various biological functions, such as transcription factor activity, transcription and transcription regulation, nucleic acid binding, sequence-specific DNA binding, p53 signaling, and post-translational modifications. Ultimately, the miRNA content of seasonally divergent FF-EVs poses as potentially relevant indicators of intrafollicular mechanisms directing seasonal transitions and their functional regulatory roles in the pre-ovulatory stage, governing subsequent ovulation.

Project description:The mammalian follicular development is a well-orchestrated and complex molecular process characterized by sequences of follicular waves that yield the ovulatory follicle. It has been reported that bi-directional communication between the growing oocyte and the surrounding somatic cells is a hallmark of mammalian follicular development, partially mediated by extracellular vesicles (EVs) present in the follicular fluid (FF). Bioactive molecules such as the miRNAs encapsulated in the EVs mediate this communication within the follicular environment. Here, we aimed to decipher the dynamics of the miRNAs in the EVs of equine FF aspirated in vivo during different stages of follicular development. Hence, all follicles ≥6 mm were ablated on Days 10–11 of the estrous cycle (ovulation = Day 0) and the new induced wave was ultrasound-monitored daily until the largest growing follicle for at least three consecutive days was randomly allocated to one of the five groups (n = 8 follicles/group) namely: predeviation (preDEV; 18–20 mm), deviation (DEV; 22–25 mm), postdeviation (postDEV; 26–29 mm), preovulatory (preOV; 30–35 mm), and impending ovulation (IMP; ~40 mm). The FF was collected from each target group by ultrasound-guided transvaginal follicle aspiration. After removal of cells, cellular debris, and larger particles, EVs were isolated from FF groups (4 biological replicates/group; 0.5 mL FF/replicate) using high-speed ultracentrifugation (120,000xg for 70 min). Isolated EVs were characterized using western blot for EV marker proteins, NanoSight Tracking Analysis, and electron microscopy. MiRNA enriched total RNA was isolated from EVs using an Exosomal RNA Isolation kit and small-RNA library preparation and RNAseq (NextSeq500; Illumina) was performed by Novogene Co, Ltd. Approximately 180 known miRNAs were detected in each group with 148 mutually found in all groups. Among the expressed miRNAs, miR-148a, miR-21, miR-192, and miR-99a were the top highly abundant miRNAs in all groups. Cluster analysis exhibited 15 different expression patterns during follicular development. Among these patterns, a group of 23 miRNAs (including miR-106b, miR-199a-5p, and miR-125a-5p) exhibited a stable expression at the preDev until the postDev stage, a significant increase at the preOV stage, and then a significant decrease at the IMP stage. This miRNA cluster has been found to target genes involved in TGF-signaling and endocytosis pathways. Another interesting pattern with 22 miRNAs (including miR-146b-5p, miR-140, and miR-143) exhibited a sharp reduction in their expression from the preDEV until the preOV stage and was predicted to target genes associated with fatty acid biosynthesis, cell cycle, and oocyte meiosis pathways. In contrast, a group of 16 miRNAs (including miR-483, miR-128, and miR-192) gradually increased in expression from preDEV to the postDEV and then downregulated at the preOV stage. This miRNA cluster targets several signaling pathways, including ErbB, thyroid, and estrogen signaling. In conclusion, this study provides greater insights into the stage-specific expression dynamics of the FF EV-miRNAs during equine follicular development, which may play a role in folliculogenesis in mares.

Dataset Information

Seasonal variation in equine follicular fluid proteome.

Publications

Seasonal variation in equine follicular fluid proteome.

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