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. 2019 Dec 16;20(24):6339.
doi: 10.3390/ijms20246339.

Metabolomic Profile of Oviductal Extracellular Vesicles Across the Estrous Cycle in Cattle

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Free PMC article

Metabolomic Profile of Oviductal Extracellular Vesicles Across the Estrous Cycle in Cattle

Julie Gatien et al. Int J Mol Sci. .
Free PMC article

Abstract

Oviductal extracellular vesicles (oEVs) have been proposed as key modulators of gamete/embryo maternal interactions. The aim of this study was to examine the metabolite content of oEVs and its regulation across the estrous cycle in cattle. Oviductal EVs were isolated from bovine oviducts ipsilateral and contralateral to ovulation at four stages of the estrous cycle (post-ovulatory stage, early and late luteal phases, and pre-ovulatory stage). The metabolomic profiling of EVs was performed by proton nuclear magnetic resonance spectroscopy (NMR). NMR identified 22 metabolites in oEVs, among which 15 were quantified. Lactate, myoinositol, and glycine were the most abundant metabolites throughout the estrous cycle. The side relative to ovulation had no effect on the oEVs' metabolite concentrations. However, levels of glucose-1-phosphate and maltose were greatly affected by the cycle stage, showing up to 100-fold higher levels at the luteal phase than at the peri-ovulatory phases. In contrast, levels of methionine were significantly higher at peri-ovulatory phases than at the late-luteal phase. Quantitative enrichment analyses of oEV-metabolites across the cycle evidenced several significantly regulated metabolic pathways related to sucrose, glucose, and lactose metabolism. This study provides the first metabolomic characterization of oEVs, increasing our understanding of the potential role of oEVs in promoting fertilization and early embryo development.

Keywords: NMR; amino acids; energy substrates; exosomes; extracellular vesicles; fallopian tube; metabolomics; oviduct.

Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Characterization of bovine oviductal extracellular vesicles (oEVs). Representative images of exosomes (30–100 nm) and microvesicles (>100 nm) in oEV preparations observed by transmission electron microscopy (TEM) across the estrus cycle (a) and Western blotting characterization of bovine oEVs for known exosomal protein markers (b). A pool of samples from four different stages was used, showing that oEVs were positive for CD81, HSP70, and ANXA1.
Figure 2
Figure 2
Comparative analysis of the oviduct EV metabolite content across the bovine estrous cycle. Principal component analysis (PCA) of metabolites measured in oEV at four different stages of the estrous cycle. S1, post-ovulation (red); S2, mid-luteal phase (blue); S3, late luteal phase (green) and S4, pre-ovulation (purple) from oviducts ipsilateral (triangles) and contralateral (round spots) to ovulation.
Figure 3
Figure 3
Differential concentrations of specific bovine oEV metabolites across the estrus cycle. Maltose intra-oEV concentrations (nmol.mg−1 of EV protein, (a) was affected by the cycle stage and side of ovulation. Glucose-1-P (b), methionine (c) and acetone (d) intra-oEV concentrations were only influenced by the stage of the estrus cycle. For (bd), ipsilateral and contralateral concentrations data were pooled as the side of ovulation did not show any significant effect on the metabolite level. (Post-ov: post-ovulatory phase; Mid-lut: mid-luteal phase; Late-lut: late lutal phase; Pre-ov: pre-ovulatory phase).
Figure 4
Figure 4
Graphical overview of the over representation analysis (ORA) for all metabolites identified in bovine oEVs generated by MetaboAnalyst 4.0 web-based software. Pathway associated metabolite sets are sorted based on fold enrichment and p value. Further details on p-values and metabolites included in each pathway are detailed in Table 4.
Figure 5
Figure 5
Graphical overview of the quantitative enrichment analysis for all metabolites quantified in bovine oEVs generated by MetaboAnalyst 4.0 web-based software. Pathway associated metabolite sets are sorted based on fold enrichment and p-value after comparison between stages. This bar chart was obtained by comparing Pre-ov vs. Late-lut stages. For each stage, ipsilateral and contralateral data were pooled. Further details on p-values and metabolites included in each pathway are detailed in Table 5.

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References

    1. Yanez-Mo M., Siljander P.R., Andreu Z., Zavec A.B., Borras F.E., Buzas E.I., Buzas K., Casal E., Cappello F., Carvalho J., et al. Biological properties of extracellular vesicles and their physiological functions. J. Extracell. Vesicles. 2015;4:27066. doi: 10.3402/jev.v4.27066. - DOI - PMC - PubMed
    1. da Silveira J.C., Veeramachaneni D.N., Winger Q.A., Carnevale E.M., Bouma G.J. Cell-secreted vesicles in equine ovarian follicular fluid contain miRNAs and proteins: A possible new form of cell communication within the ovarian follicle. Biol. Reprod. 2012;86:71. doi: 10.1095/biolreprod.111.093252. - DOI - PubMed
    1. Ng Y.H., Rome S., Jalabert A., Forterre A., Singh H., Hincks C.L., Salamonsen L.A. Endometrial exosomes/microvesicles in the uterine microenvironment: A new paradigm for embryo-endometrial cross talk at implantation. PLoS ONE. 2013;8:e58502 doi: 10.1371/journal.pone.0058502. - DOI - PMC - PubMed
    1. Lopera-Vasquez R., Hamdi M., Maillo V., Gutierrez-Adan A., Bermejo-Alvarez P., Ramirez M.A., Yanez-Mo M., Rizos D. Effect of bovine oviductal extracellular vesicles on embryo development and quality in vitro. Reproduction. 2017;153:461–470. doi: 10.1530/REP-16-0384. - DOI - PubMed
    1. Alminana C., Corbin E., Tsikis G., Alcantara-Neto A.S., Labas V., Reynaud K., Galio L., Uzbekov R., Garanina A.S., Druart X., et al. Oviduct extracellular vesicles protein content and their role during oviduct-embryo cross-talk. Reproduction. 2017;154:153–168. doi: 10.1530/REP-17-0054. - DOI - PubMed
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