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Open AccessArticle

Spatial and Pregnancy-Related Changes in the Protein, Amino Acid, and Carbohydrate Composition of Bovine Oviduct Fluid

1
Department of Animal Reproduction, National Institute for Agricultural and Food Research and Technology (INIA), Ctra. de la Coruña KM 5.9, 28040 Madrid, Spain
2
School of Agriculture and Food Science, University College Dublin, Belfield, Dublin 4, Ireland
3
Department of Cell Biology and Histology, Faculty of Medicine, Instituto Murciano de Investigación Biosanitaria (IMIB-Arrixaca), University of Murcia, 30100 Murcia, Spain
4
Faculty of Veterinary and Zootechnics, Autonomous University of Sinaloa, Culiacan 80246, Mexico
5
Department of Research, Animal Reproduction Biotechnology (ARBiotech), Culiacan 80015, Mexico
6
Molecular Biology Section, SAI, University of Murcia, 30100 Murcia, Spain
7
Center for Atherothrombosis and Metabolic Disease, Hull York Medical School, University of Hull, Hull HU3 2JZ, UK
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(5), 1681; https://doi.org/10.3390/ijms21051681
Received: 12 February 2020 / Revised: 27 February 2020 / Accepted: 27 February 2020 / Published: 29 February 2020
(This article belongs to the Special Issue Embryo-Maternal Interactions Underlying Reproduction in Mammals)
Knowledge of how the biochemical composition of the bovine oviduct is altered due to the oviduct anatomy or the presence of an embryo is lacking. Thus, the aim of this study was to assess the effect of (І) oviduct anatomy and (ІІ) embryo presence on oviductal fluid (OF) protein, amino acid, and carbohydrate composition. Cross-bred beef heifers (n = 19) were synchronized and those in standing estrus were randomly allocated to a cyclic (non-bred) or pregnant (artificially inseminated) group. All heifers were slaughtered on Day 3 after estrus. The oviducts ipsilateral to the corpus luteum from each animal were isolated, straightened and cut, separating ampulla and isthmus. Each portion was flushed with 500 µl of PBS enabling recovery of the oocyte/embryo. Recovered unfertilized oocytes (cyclic group) and embryos (8-cell embryos; pregnant group) were located in the isthmus of the oviduct. Samples of flushing medium from the isthmus and ampulla were used for proteomic (n = 2 per group), amino acid (n = 5), and carbohydrate (n = 5) analysis. For proteomic analysis, total protein from cyclic and pregnant samples were labelled with different cyanine fluorescent probes and separated according to the isoelectric point using immobilized pH gradient strips (pH 3–10, 17 cm, Protean® IEF cell system, Bio Rad). Second dimension was performed in a polyacrylamide gel (12%) in the presence of SDS using a Protean II XL system (Bio Rad). Images were obtained with a Typhoon 9410 scanner and analyzed with Progenesis SameSpots software v 4.0. Amino acid content in the OF was determined by high performance liquid chromatography (HPLC). Glucose, lactate, and pyruvate were quantified using microfluorometric enzyme-linked assays. For the proteomic assessment, the results of the image analysis were compared by ANOVA. For both amino acid and carbohydrate analyses, statistical analysis was carried out by 2-way ANOVA with the Holm-Sidak nonparametric post hoc analysis. On Day 3 post-estrus, OF composition varied based on (І) anatomical region, where isthmic metabolites were present in lower (i.e., lactate, glycine, and alanine) or higher (i.e., arginine) concentrations compared to the ampulla; and (ІІ) embryo presence, which was correlated with greater, arginine, phosphoglycerate kinase 1, serum albumin, α-1-antiproteinase and [email protected] protein concentrations. In conclusion, data indicate that the composition of bovine OF is anatomically dynamic and influenced by the presence of an early embryo. View Full-Text
Keywords: oviduct fluid; pregnancy; proteome; metabolome; bovine oviduct fluid; pregnancy; proteome; metabolome; bovine
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MDPI and ACS Style

Rodríguez-Alonso, B.; Maillo, V.; Acuña, O.S.; López-Úbeda, R.; Torrecillas, A.; Simintiras, C.A.; Sturmey, R.; Avilés, M.; Lonergan, P.; Rizos, D. Spatial and Pregnancy-Related Changes in the Protein, Amino Acid, and Carbohydrate Composition of Bovine Oviduct Fluid. Int. J. Mol. Sci. 2020, 21, 1681. https://doi.org/10.3390/ijms21051681

AMA Style

Rodríguez-Alonso B, Maillo V, Acuña OS, López-Úbeda R, Torrecillas A, Simintiras CA, Sturmey R, Avilés M, Lonergan P, Rizos D. Spatial and Pregnancy-Related Changes in the Protein, Amino Acid, and Carbohydrate Composition of Bovine Oviduct Fluid. International Journal of Molecular Sciences. 2020; 21(5):1681. https://doi.org/10.3390/ijms21051681

Chicago/Turabian Style

Rodríguez-Alonso, Beatriz; Maillo, Veronica; Acuña, Omar S.; López-Úbeda, Rebeca; Torrecillas, Alejandro; Simintiras, Constantine A.; Sturmey, Roger; Avilés, Manuel; Lonergan, Patrick; Rizos, Dimitrios. 2020. "Spatial and Pregnancy-Related Changes in the Protein, Amino Acid, and Carbohydrate Composition of Bovine Oviduct Fluid" Int. J. Mol. Sci. 21, no. 5: 1681. https://doi.org/10.3390/ijms21051681

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