The Proteome of Equine Oviductal Fluid Varies Before and After Ovulation: A Comparative Study.
Abstract: Equine fertilization cannot be performed in the laboratory as equine spermatozoa do not cross the oocyte's zona pellucida . Hence, a more profound study of equine oviductal fluid (OF) composition at the pre-ovulatory and post-ovulatory stages could help in understanding what components are required to achieve fertilization in horses. Our work aimed to elucidate the proteomic composition of equine OF at both stages. To do this, OF was obtained postmortem from oviducts of slaughtered mares ipsilateral to a pre-ovulatory follicle ( = 4) or a recent ovulation ( = 4); the samples were kept at -80°C until analysis. After protein extraction and isobaric tags for relative and absolute quantification (iTRAQ) labeling, the samples were analyzed by nano-liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The analysis of the spectra resulted in the identification of a total of 1,173 proteins present in pre-ovulatory and post-ovulatory samples; among these, 691 were unique for . Proteins from post-ovulatory oviductal fluid were compared with the proteins from pre-ovulatory oviductal fluid and were categorized as upregulated (positive log fold change) or downregulated (negative log fold change). Fifteen proteins were found to be downregulated in the post-ovulatory fluid and 156 were upregulated in the post-ovulatory OF compared to the pre-ovulatory fluid; among the upregulated proteins, 87 were included in the pathway. The identified proteins were related to , and , among others. Our data reveal consistent differences in the proteome of equine OF prior to and after ovulation, helping to increase our understanding in the factors that promote fertilization and early embryo development in horses.
Copyright © 2021 Fernández-Hernández, Marinaro, Sánchez-Calabuig, García-Marín, Bragado, González-Fernández and Macías-García.
Publication Date: 2021-08-05 PubMed ID: 34422946PubMed Central: PMC8375304DOI: 10.3389/fvets.2021.694247Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This study focused on identifying the differences in protein composition of the oviductal fluid (OF) in horses before and after ovulation to understand the factors needed for successful fertilization in horses.
Objective and Rationale
- The research focused on understanding the processes and factors necessary for equine fertilization, which has proven to be challenging in a lab setting due to the inability of horse sperm cells to cross the outer layer of the ovum, zona pellucida. As such, significant attention was placed on evaluating the OF at both pre- and post-ovulatory stages.
Methods and Experimentation
- The researchers acquired OF samples post-mortem from horses that were either alongside a pre-ovulatory follicle or a recent ovulation. These samples were preserved at -80°C until the analysis phase.
- Protein extraction was then carried out on the samples. The extracted proteins were quantified using, isobaric tags for relative and absolute quantification (iTRAQ) labelling methodology.
- Further analysis was performed by nano-liquid chromatography combined with tandem mass spectrometry (LC-MS/MS). Together, these steps allowed the researchers to identify a total of 1,173 proteins in both pre-ovulatory and post-ovulatory samples.
Findings
- Out of the total proteins identified, 691 were unique. A comparison of proteins in post-ovulatory OF with that of pre-ovulatory OF showed that some proteins were upregulated or downregulated.
- More specifically, 15 proteins were found to have downregulated (decreased) in the post-ovulatory fluid compared to the pre-ovulatory fluid. On the other hand, 156 proteins were upregulated (increased) in the post-ovulatory OF relative to the pre-ovulatory fluid. Among these upregulated proteins, 87 were part of the unique set of 691 proteins.
Significance
- This research elucidated the differences in protein structure of equine OF before and after ovulation. The findings provide a broader understanding of the proteins and factors that potentially aid fertilization and early embryo development in horses.
Cite This Article
APA
Fernández-Hernández P, Marinaro F, Sánchez-Calabuig MJ, García-Marín LJ, Bragado MJ, González-Fernández L, Macías-García B.
(2021).
The Proteome of Equine Oviductal Fluid Varies Before and After Ovulation: A Comparative Study.
Front Vet Sci, 8, 694247.
https://doi.org/10.3389/fvets.2021.694247 Publication
Researcher Affiliations
- Research Group of Intracellular Signaling and Technology of Reproduction (Research Institute INBIO G+C), University of Extremadura, Cáceres, Spain.
- Department of Animal Medicine, Faculty of Veterinary Sciences, University of Extremadura, Cáceres, Spain.
- Stem Cell Therapy Unit, Jesús Usón Minimally Invasive Surgery Centre, Cáceres, Spain.
- Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, University Complutense of Madrid, Madrid, Spain.
- Research Group of Intracellular Signaling and Technology of Reproduction (Research Institute INBIO G+C), University of Extremadura, Cáceres, Spain.
- Department of Physiology, Faculty of Veterinary Sciences, University of Extremadura, Cáceres, Spain.
- Research Group of Intracellular Signaling and Technology of Reproduction (Research Institute INBIO G+C), University of Extremadura, Cáceres, Spain.
- Department of Biochemistry and Molecular Biology and Genetics, Faculty of Veterinary Sciences, University of Extremadura, Cáceres, Spain.
- Research Group of Intracellular Signaling and Technology of Reproduction (Research Institute INBIO G+C), University of Extremadura, Cáceres, Spain.
- Department of Biochemistry and Molecular Biology and Genetics, Faculty of Veterinary Sciences, University of Extremadura, Cáceres, Spain.
- Research Group of Intracellular Signaling and Technology of Reproduction (Research Institute INBIO G+C), University of Extremadura, Cáceres, Spain.
- Department of Animal Medicine, Faculty of Veterinary Sciences, University of Extremadura, Cáceres, Spain.
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
References
This article includes 57 references
- Galli C, Lazzari G. The manipulation of gametes and embryos in farm animals.. Reprod Domest Anim 2008 Jul;43 Suppl 2:1-7.
- Lopera-Vasquez R, Hamdi M, Maillo V, Lloreda V, Coy P, Gutierrez-Adan A, Bermejo-Alvarez P, Rizos D. Effect of bovine oviductal fluid on development and quality of bovine embryos produced in vitro.. Reprod Fertil Dev 2017 Mar;29(3):621-629.
- THIBAULT C, DAUZIER L, WINTENBERGER S. [Cytological study of fecundation in vitro of rabbit ovum].. C R Seances Soc Biol Fil 1954 May;148(9-10):789-90.
- CHANG MC. Fertilizing capacity of spermatozoa deposited into the fallopian tubes.. Nature 1951 Oct 20;168(4277):697-8.
- AUSTIN CR. Observations on the penetration of the sperm in the mammalian egg.. Aust J Sci Res B 1951 Nov;4(4):581-96.
- Coy P, Yanagimachi R. The common and species-specific roles of oviductal proteins in mammalian fertilization and embryo development.. BioScience (2015) 65:973–84.
- Gardner DK, Leese HJ. Concentrations of nutrients in mouse oviduct fluid and their effects on embryo development and metabolism in vitro.. J Reprod Fertil 1990 Jan;88(1):361-8.
- Leemans B, Gadella BM, Stout TA, De Schauwer C, Nelis H, Hoogewijs M, Van Soom A. Why doesn't conventional IVF work in the horse? The equine oviduct as a microenvironment for capacitation/fertilization.. Reproduction 2016 Dec;152(6):R233-R245.
- Leemans B, Stout TAE, De Schauwer C, Heras S, Nelis H, Hoogewijs M, Van Soom A, Gadella BM. Update on mammalian sperm capacitation: how much does the horse differ from other species?. Reproduction 2019 May;157(5):R181-R197.
- Choi YH, Okada Y, Hochi S, Braun J, Sato K, Oguri N. In vitro fertilization rate of horse oocytes with partially removed zonae.. Theriogenology 1994 Oct;42(5):795-802.
- González-Fernández L, Sánchez-Calabuig MJ, Calle-Guisado V, García-Marín LJ, Bragado MJ, Fernández-Hernández P, Gutiérrez-Adán A, Macías-García B. Stage-specific metabolomic changes in equine oviductal fluid: New insights into the equine fertilization environment.. Theriogenology 2020 Feb;143:35-43.
- Hinrichs K, Love CC, Brinsko SP, Choi YH, Varner DD. In vitro fertilization of in vitro-matured equine oocytes: effect of maturation medium, duration of maturation, and sperm calcium ionophore treatment, and comparison with rates of fertilization in vivo after oviductal transfer.. Biol Reprod 2002 Jul;67(1):256-62.
- Lamy J, Gatien J, Dubuisson F, Nadal-Desbarats L, Salvetti P, Mermillod P, Saint-Dizier M. Metabolomic profiling of bovine oviductal fluid across the oestrous cycle using proton nuclear magnetic resonance spectroscopy.. Reprod Fertil Dev 2018 Jun;30(7):1021-1028.
- Bauersachs S, Blum H, Mallok S, Wenigerkind H, Rief S, Prelle K, Wolf E. Regulation of ipsilateral and contralateral bovine oviduct epithelial cell function in the postovulation period: a transcriptomics approach.. Biol Reprod 2003 Apr;68(4):1170-7.
- Maillo V, de Frutos C, O'Gaora P, Forde N, Burns GW, Spencer TE, Gutierrez-Adan A, Lonergan P, Rizos D. Spatial differences in gene expression in the bovine oviduct.. Reproduction 2016 Jul;152(1):37-46.
- Hunter RH. Components of oviduct physiology in eutherian mammals.. Biol Rev Camb Philos Soc 2012 Feb;87(1):244-55.
- Avilés M, Gutiérrez-Adán A, Coy P. Oviductal secretions: will they be key factors for the future ARTs?. Mol Hum Reprod 2010 Dec;16(12):896-906.
- Saint-Dizier M, Schoen J, Chen S, Banliat C, Mermillod P. Composing the Early Embryonic Microenvironment: Physiology and Regulation of Oviductal Secretions.. Int J Mol Sci 2019 Dec 28;21(1).
- Smits K, Nelis H, Van Steendam K, Govaere J, Roels K, Ververs C, Leemans B, Wydooghe E, Deforce D, Van Soom A. Proteome of equine oviducal fluid: effects of ovulation and pregnancy.. Reprod Fertil Dev 2017 Jun;29(6):1085-1095.
- Ramos-Fernández A, Paradela A, Navajas R, Albar JP. Generalized method for probability-based peptide and protein identification from tandem mass spectrometry data and sequence database searching.. Mol Cell Proteomics 2008 Sep;7(9):1748-54.
- Lopez-Serra P, Marcilla M, Villanueva A, Ramos-Fernandez A, Palau A, Leal L, Wahi JE, Setien-Baranda F, Szczesna K, Moutinho C, Martinez-Cardus A, Heyn H, Sandoval J, Puertas S, Vidal A, Sanjuan X, Martinez-Balibrea E, Viñals F, Perales JC, Bramsem JB, Ørntoft TF, Andersen CL, Tabernero J, McDermott U, Boxer MB, Vander Heiden MG, Albar JP, Esteller M. A DERL3-associated defect in the degradation of SLC2A1 mediates the Warburg effect.. Nat Commun 2014 Apr 3;5:3608.
- Huang da W, Sherman BT, Lempicki RA. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources.. Nat Protoc 2009;4(1):44-57.
- Huang da W, Sherman BT, Lempicki RA. Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists.. Nucleic Acids Res 2009 Jan;37(1):1-13.
- Reimand J, Kull M, Peterson H, Hansen J, Vilo J. g:Profiler--a web-based toolset for functional profiling of gene lists from large-scale experiments.. Nucleic Acids Res 2007 Jul;35(Web Server issue):W193-200.
- Lamy J, Labas V, Harichaux G, Tsikis G, Mermillod P, Saint-Dizier M. Regulation of the bovine oviductal fluid proteome.. Reproduction 2016 Dec;152(6):629-644.
- Capra E, Lange-Consiglio A. The Biological Function of Extracellular Vesicles during Fertilization, Early Embryo-Maternal Crosstalk and Their Involvement in Reproduction: Review and Overview.. Biomolecules 2020 Nov 4;10(11).
- Binelli M, Gonella-Diaza AM, Mesquita FS, Membrive CMB. Sex Steroid-Mediated Control of Oviductal Function in Cattle.. Biology (Basel) 2018 Feb 2;7(1).
- Saint-Dizier M, Marnier C, Tahir MZ, Grimard B, Thoumire S, Chastant-Maillard S, Reynaud K. OVGP1 is expressed in the canine oviduct at the time and place of oocyte maturation and fertilization.. Mol Reprod Dev 2014 Nov;81(11):972-82.
- Van Winkle LJ. Amino acid transport regulation and early embryo development.. Biol Reprod 2001 Jan;64(1):1-12.
- Hotamisligil GS, Bernlohr DA. Metabolic functions of FABPs--mechanisms and therapeutic implications.. Nat Rev Endocrinol 2015 Oct;11(10):592-605.
- Del Collado M, da Silveira JC, Sangalli JR, Andrade GM, Sousa LRDS, Silva LA, Meirelles FV, Perecin F. Fatty Acid Binding Protein 3 And Transzonal Projections Are Involved In Lipid Accumulation During In Vitro Maturation Of Bovine Oocytes.. Sci Rep 2017 Jun 1;7(1):2645.
- Smits K, Goossens K, Van Soom A, Govaere J, Hoogewijs M, Peelman LJ. In vivo-derived horse blastocysts show transcriptional upregulation of developmentally important genes compared with in vitro-produced horse blastocysts.. Reprod Fertil Dev 2011;23(2):364-75.
- Song GX, Shen YH, Liu YQ, Sun W, Miao LP, Zhou LJ, Liu HL, Yang R, Kong XQ, Cao KJ, Qian LM, Sheng YH. Overexpression of FABP3 promotes apoptosis through inducing mitochondrial impairment in embryonic cancer cells.. J Cell Biochem 2012 Dec;113(12):3701-8.
- Budna J, Celichowski P, Knap S, Jankowski M, Magas M, Nawrocki MJ. Fatty acids related genes expression undergo substantial changes in porcine oviductal epithelial cells during long-term primary culture.. Med J Cell Biol (2018) 6:39–47.
- Walter J, Huwiler F, Fortes C, Grossmann J, Roschitzki B, Hu J, Naegeli H, Laczko E, Bleul U. Analysis of the equine "cumulome" reveals major metabolic aberrations after maturation in vitro.. BMC Genomics 2019 Jul 17;20(1):588.
- Rodriguez-Martinez H, Tienthai P, Atikuzzaman M, Vicente-Carrillo A, Rubér M, Alvarez-Rodriguez M. The ubiquitous hyaluronan: Functionally implicated in the oviduct?. Theriogenology 2016 Jul 1;86(1):182-6.
- Darr CR, Varner DD, Teague S, Cortopassi GA, Datta S, Meyers SA. Lactate and Pyruvate Are Major Sources of Energy for Stallion Sperm with Dose Effects on Mitochondrial Function, Motility, and ROS Production.. Biol Reprod 2016 Aug;95(2):34.
- González-Fernández L, Sánchez-Calabuig MJ, Alves MG, Oliveira PF, Macedo S, Gutiérrez-Adán A, Rocha A, Macías-García B. Expanded equine cumulus-oocyte complexes exhibit higher meiotic competence and lower glucose consumption than compact cumulus-oocyte complexes.. Reprod Fertil Dev 2018 Jan;30(2):297-306.
- Lewis N, Hinrichs K, Leese HJ, McG Argo C, Brison DR, Sturmey R. Energy metabolism of the equine cumulus oocyte complex during in vitro maturation.. Sci Rep 2020 Feb 26;10(1):3493.
- Steinberger B, Yu H, Brodmann T, Milovanovic D, Reichart U, Besenfelder U, Artemenko K, Razzazi-Fazeli E, Brem G, Mayrhofer C. Semen modulated secretory activity of oviductal epithelial cells is linked to cellular proteostasis network remodeling: Proteomic insights into the early phase of interaction in the oviduct in vivo.. J Proteomics 2017 Jun 23;163:14-27.
- Yu H, Hackenbroch L, Meyer FRL, Reiser J, Razzazi-Fazeli E, Nöbauer K, Besenfelder U, Vogl C, Brem G, Mayrhofer C. Identification of Rabbit Oviductal Fluid Proteins Involved in Pre-Fertilization Processes by Quantitative Proteomics.. Proteomics 2019 Mar;19(5):e1800319.
- Saewu A, Kadunganattil S, Raghupathy R, Kongmanas K, Diaz-Astudillo P, Hermo L, Tanphaichitr N. Clusterin in the mouse epididymis: possible roles in sperm maturation and capacitation.. Reproduction 2017 Dec;154(6):867-880.
- Smits K, Willems S, Van Steendam K, Van De Velde M, De Lange V, Ververs C, Roels K, Govaere J, Van Nieuwerburgh F, Peelman L, Deforce D, Van Soom A. Proteins involved in embryo-maternal interaction around the signalling of maternal recognition of pregnancy in the horse.. Sci Rep 2018 Mar 27;8(1):5249.
- Hemachand T, Gopalakrishnan B, Salunke DM, Totey SM, Shaha C. Sperm plasma-membrane-associated glutathione S-transferases as gamete recognition molecules.. J Cell Sci 2002 May 15;115(Pt 10):2053-65.
- Soleilhavoup C, Riou C, Tsikis G, Labas V, Harichaux G, Kohnke P, Reynaud K, de Graaf SP, Gerard N, Druart X. Proteomes of the Female Genital Tract During the Oestrous Cycle.. Mol Cell Proteomics 2016 Jan;15(1):93-108.
- Liang X, Jin Y, Wang H, Meng X, Tan Z, Huang T, Fan S. Transgelin 2 is required for embryo implantation by promoting actin polymerization.. FASEB J 2019 Apr;33(4):5667-5675.
- Breitbart H, Cohen G, Rubinstein S. Role of actin cytoskeleton in mammalian sperm capacitation and the acrosome reaction.. Reproduction 2005 Mar;129(3):263-8.
- Puts GS, Leonard MK, Pamidimukkala NV, Snyder DE, Kaetzel DM. Nuclear functions of NME proteins.. Lab Invest 2018 Feb;98(2):211-218.
- Almiñana C, Tsikis G, Labas V, Uzbekov R, da Silveira JC, Bauersachs S, Mermillod P. Deciphering the oviductal extracellular vesicles content across the estrous cycle: implications for the gametes-oviduct interactions and the environment of the potential embryo.. BMC Genomics 2018 Aug 22;19(1):622.
- Sommer A, Christensen E, Schwenger S, Seul R, Haas D, Olbrich H, Omran H, Sass JO. The molecular basis of aminoacylase 1 deficiency.. Biochim Biophys Acta 2011 Jun;1812(6):685-90.
- Gardner DK, Wale PL. Analysis of metabolism to select viable human embryos for transfer.. Fertil Steril 2013 Mar 15;99(4):1062-72.
- Hess AP, Talbi S, Hamilton AE, Baston-Buest DM, Nyegaard M, Irwin JC, Barragan F, Kruessel JS, Germeyer A, Giudice LC. The human oviduct transcriptome reveals an anti-inflammatory, anti-angiogenic, secretory and matrix-stable environment during embryo transit.. Reprod Biomed Online 2013 Oct;27(4):423-35.
- Onken J, Moeckel S, Leukel P, Leidgens V, Baumann F, Bogdahn U, Vollmann-Zwerenz A, Hau P. Versican isoform V1 regulates proliferation and migration in high-grade gliomas.. J Neurooncol 2014 Oct;120(1):73-83.
- Ricciardelli C, Russell DL, Ween MP, Mayne K, Suwiwat S, Byers S, Marshall VR, Tilley WD, Horsfall DJ. Formation of hyaluronan- and versican-rich pericellular matrix by prostate cancer cells promotes cell motility.. J Biol Chem 2007 Apr 6;282(14):10814-25.
- Lamy J, Nogues P, Combes-Soia L, Tsikis G, Labas V, Mermillod P, Druart X, Saint-Dizier M. Identification by proteomics of oviductal sperm-interacting proteins.. Reproduction 2018 May;155(5):457-466.
- Bernabò N, Ordinelli A, Di Agostino R, Mattioli M, Barboni B. Network analyses of sperm-egg recognition and binding: ready to rethink fertility mechanisms?. OMICS 2014 Dec;18(12):740-53.
- Bromfield EG, Nixon B. The function of chaperone proteins in the assemblage of protein complexes involved in gamete adhesion and fusion processes.. Reproduction 2013 Feb;145(2):R31-42.
Citations
This article has been cited 3 times.- Lawson EF, Ghosh A, Blanch V, Grupen CG, Aitken RJ, Lim R, Drury HR, Baker MA, Gibb Z, Tanwar PS. Establishment and characterization of oviductal organoids from farm and companion animals†. Biol Reprod 2023 Jun 9;108(6):854-865.
- Mahé C, Lavigne R, Com E, Pineau C, Locatelli Y, Zlotkowska AM, Almiñana C, Tsikis G, Mermillod P, Schoen J, Saint-Dizier M. Spatiotemporal profiling of the bovine oviduct fluid proteome around the time of ovulation. Sci Rep 2022 Mar 9;12(1):4135.
- Lösle M, Lin CW, Beil-Wagner J, Aebi M, Buch T. Comparison of pregnant mare serum gonadotropin products with surprising differences in protein content. Sci Rep 2025 Feb 25;15(1):6824.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
