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Singina G.N., Shedova E.N., Uzbekov R., Chinarov R.Yu., Lukanina V.A., Uzbekova S. Presence of follicular fluid extracellular vesiclesduring in vitro maturation of donor cow (Bos taurus) oocytes increases their ability to in vitro embryo development. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2023, Vol. 58, № 6, p. 1100-1111.
(how to cite this article)

doi: 10.15389/agrobiology.2023.6.1100eng

UDC: 636.2:591.39

Acknowledgements:
Supported financially by the Russian Science Foundation, project No. 19-16-00115-П

 

PRESENCE OF FOLLICULAR FLUID EXTRACELLULAR VESICLES DURING in vitro MATURATION OF DONOR COW (Bos taurus) OOCYTES INCREASES THEIR ABILITY TO in vitro EMBRYO DEVELOPMENT

G.N. Singina1 ✉, E.N. Shedova1, R. Uzbekov2, 3,
R.Yu. Chinarov1, V.A. Lukanina1, S. Uzbekova4

1Ernst Federal Research Center for Animal Husbandry, 60, pos. Dubrovitsy, Podolsk District, Moscow Province, 142132 Russia, e-mail g_singina@mail.ru (✉ corresponding author), shedvek@yandex.ru, roman_chinarov@mail.ru, kristybatle@gmail.com;
2Laboratory of Cell Biology and Electron Microscopy, Faculty of Medicine, University of Tours, 37032 Tours, France, 10 bd Tonnelle, e-mail rustem.uzbekov@univ-tours.fr;
3Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, GSP-1, 1-73, Leninskiye Gory, Moscow, 119234 Russia;
4Physiology of Reproduction and Behavior unit, INRAE, CNRS, IFCE, Tours University, 37380 Nouzilly, France, e-mail svetlana.uzbekova@inrae.fr

ORCID:
Singina G.N. orcid.org/0000-0003-0198-9757
Chinarov R.Yu. orcid.org/0000-0001-6511-5341
Shedova E.N. orcid.org/0000-0002-9642-2384
Lukanina V.A. orcid.org/0000-0003-4744-7873
Uzbekov R. orcid.org/0000-0002-9336-5484
Uzbekova S. orcid.org/0000-0002-6989-6904

Final revision received November 03, 2023
Accepted November 30, 2023

In vitro embryo production (IVEP) technology from the oocytes isolated by transvaginal follicle puncture (ovum-pickup, OPU) is widely used to generate a number of descendants from the most valuable donor cows and becomes a routine in cattle breeding programs to replicate and conserve precious genotypes. To increase the efficiency of OPU/IVEP-technology, in this work for the first time, bovine OPU-oocytes were matured in the presence of extracellular vesicles (EVs) from follicular fluid (FF), and the ability of treated oocytes to IVEP after in vitro fertilization was investigated. The aim was to study the effects of FF EVs on oocyte maturation and capacity to develop up to blastocyst of OPU-oocytes, as well as to resistance of such blastocysts to freezing. EVs were obtained from FF by differential centrifugations followed by ultracentrifugation at 100,000 g. The preparations were analyzed by transmission electron microscopy that confirmed the presence of exosome-size EVs. Oocyte donors were sexually mature Yaroslavl breed heifers (n = 6) following natural cycle. OPU was performed twice a week. The isolated oocytes were in vitro matured in TC-199 medium, supplemented by fetal bull serum (10 %), follicle stimulating and luteinizing (10 mg/ml) hormones, epidermal growth factor (10 ng/ml) in absence (control) or presence of EVs (experiment). Vesicular preparations were added to the in vitro maturation (IVM) medium in the physiological concentration (EVs isolated from 1 ml of FF per 1 ml of medium). After 24 hours of IVM, the oocytes were subjected to in-vitro fertilization and in vitro embryo culture. At day 3 after fertilization, oocyte morphology was checked, and at day 7 of culture, a number of embryos developed to blastocyst (Bl) was determined. The resulting Bl were frozen and stored for some time at -196 °С. Then the blastocysts were thawed and in vitro cultured until hatching to determine their viability. A total of 10 independent experiments were performed, 57 and 56 oocytes were analyzed in control and treatment, respectively. No impact of experimental conditions on nuclear maturation rate was evidenced. The mature oocytes were similar in control and EVs-treated groups and accounted for 90.4±5.6 % and 94.3±3.1 %, respectively. In addition, the presence of EVs during IVM did not change oocyte cleavage rate after vitro fertilization, 78.6±7.3 % and 86.7±4.9 % in control and EV-treated groups, respectively. However, beneficial effect of EVs on blastocyst rate was found. In control, 26.6±5.8 % of OPU oocytes developed to Bl. EVs added during IVM increased blastocyst rate to 41.2±3.2 % (p < 0.05). EVs also tended to increase cryoresistance of resulting blastocysts. In control, blastocyst hatching rate after thawing and short-term culture was 29.1±8.8 % and increased to 53.3±9.2% in the EVs group. Thus, addition of EVs from cow follicular fluid during IVM culture increases oocyte quality and, consequently, their competence to embryo development after in vitro fertilization. Therefore, using of follicular fluid EVs during in vitro maturation of OPU oocytes can improve the efficiency of OPU/IVEP technologies in cattle.

Keywords: extracellular vesicles, bovine follicular fluid, bovine oocytes, in vitro maturation, oocyte aging, embryo development.

 

REFERENCES

  1. Mapletoft R.J. History and perspectives on bovine embryo transfer. Animal Reproduction, 2013, 10(3): 168-173.
  2. Sirard M.A. 40 years of bovine IVF in the new genomic selection context. Reproduction, 2018, 156(1): 1-7 CrossRef
  3. Qi M., Yao Y., Ma H., Wang J., Zhao X., Liu L., Tang X., Zhang L., Zhang S., Sun F. Transvaginal ultrasound-guided Ovum Pick-up (OPU) in cattle. Journal of Biomaterials and Tissue Engineering, 2013, 18: Article 118 CrossRef
  4. Boni R. Ovum pick-up in cattle: A 25 yr retrospective analysis. Animal Reproduction, 2012, 9(3): 362-369.
  5. Viana J. 2019 Statistics of embryo production and transfer in domestic farm animals. Embryo Technology Newsletter, 2020, 38(4): 7-26.
  6. Viana J. 2021 Statistics of embryo production and transfer in domestic farm animals. Embryo Technology Newsletter, 2022, 38(4): 22-40.
  7. Ferré L.B., Kjelland M.E., Strøbech L.B., Hyttel P., Mermillod P., Ross P.J. Review: Recent advances in bovine in vitro embryo production: reproductive biotechnology history and methods. Animal, 2020, 14(5): 991-1004 CrossRef
  8. Sanches B.V., Zangirolamo A.F., Seneda M.M. Intensive use of IVF by large-scale dairy programs. Animal Reproduction, 2019, 16(3): 394-401 CrossRef
  9. Ashry M., Smith G.W. Application of embryo transfer using in vitro produced embryos: intrinsic factors affecting efficiency. Cattle Practice: Journal of the British Cattle Veterinary Association, 2015, 23(Pt. 1): 1-8.
  10. Ferré L.B., Kjelland M.E., Taiyeb A.M., Campos-Chillon F., Ross P.J. Recent progress in bovine in vitro-derived embryo cryotolerance: impact of in vitro culture systems, advances in cryopreservation and future considerations. Reproduction in Domestic Animals, 2020, 55(6): 659-676 CrossRef
  11. Marsico T.V., de Camargo J., Valente R.S., Sudano M.J. Embryo competence and cryosurvival: Molecular and cellular features. Animal Reproduction, 2019, 16(3): 423-439 CrossRef
  12. Thompson J.G., Lane M., Gilchrist R.B. Metabolism of the bovine cumulus-oocyte complex and influence on subsequent developmental competence. Society of Reproduction and Fertility Supplement, 2007, 64: 179-190 CrossRef
  13. Wrenzycki C., Stinshoff H. Maturation environment and impact on subsequent developmental competence of bovine oocytes. Reproduction in Domestic Animals, 2013, 48(1): 38-43 CrossRef
  14. Mingoti G.Z., Castro V.S., Méo S.C., Sά Barreto L.S., Garcia J.M. The effects of macromolecular and serum supplements and oxygen tension during bovine in vitro procedures on kinetics of oocyte maturation and embryo development. In Vitro Cellular & Developmental Biology. Animal, 2011, 47: 361-367 CrossRef
  15. Lonergan P., Fair T. Maturation of oocytes in vitro. Annual Review of Animal Biosciences, 2016, 4: 255-268 CrossRef
  16. Luciano A.M., Franciosi F., Barros, R.G., Dieci C., Lodde V. The variable success of in vitro maturation: can we do better? Animal Reproduction, 2018, 15: 727-736 CrossRef
  17. Peixoto de Souza V., Jensen J., Whitler W., Estill C.T., Bishop C.V. Increasing vitamin D levels to improve fertilization rates in cattle. Journal of Animal Science, 2022, 100(7): Article skac168 CrossRef
  18. Raposo G., Stoorvogel W. Extracellular vesicles: exosomes, microvesicles, and friends. Journal of Cell Biology, 2013, 200(4): 373-383 CrossRef
  19. Record M., Carayon K., Poirot M., Silvente-Poirot S. Exosomes as new vesicular lipid transporters involved in cell-cell communication and various pathophysiologies. Biochimica et Biophysica Acta, 2014, 1841(1): 108-120 CrossRef
  20. Uzbekova S., Almiñana C., Labas V., Teixeira-Gomes A.P., Combes-Soia L., Tsikis G., Carvalho A.V., Uzbekov R., Singina G. Protein cargo of extracellular vesicles from bovine follicular fluid and analysis of their origin from different ovarian cells. Frontiers in Veterinary Science, 2020, 7: Article 584948 CrossRef
  21. Maugrion E., Shedova E.N, Uzbekov R., Teixeira-Gomes A.P., Labas V., Tomas D., Banliat C., Singina G.N., Uzbekova S. Extracellular vesicles contribute to the difference in lipid composition between ovarian follicles of different size revealed by mass spectrometry imaging. Metabolites, 2023, 13: Article 1001 CrossRef
  22. Tesfaye D., Hailay T., Salilew-Wondim D., Hoelker M., Bitseha S., Gebremedhn S. Extracellular vesicle mediated molecular signaling in ovarian follicle: Implication for oocyte developmental competence. Theriogenology, 2020, 150: 70-74 CrossRef
  23. Hailay T., Hoelker M., Poirier M., Gebremedhn S., Rings F., Saeed-Zidane M., Salilew-Wondim D., Dauben C., Tholen E., Neuhoff C., Schellander K., Tesfaye D. Extracellular vesicle-coupled miRNA profiles in follicular fluid of cows with divergent post-calving metabolic status. Scientific Reproduction, 2019, 9(1): Article 12851 CrossRef
  24. Asaadi A., Dolatabad N.A., Atashi H., Raes A., Van Damme P., Hoelker M., Hendrix A., Pascottini O.B., Van Soom A., Kafi M., Pavani K.C. Extracellular vesicles from follicular and ampullary fluid isolated by density gradient ultracentrifugation improve bovine embryo development and quality. International Journal of Molecular Sciences, 2021, 22(2): Article 578 CrossRef
  25. Rodrigues T.A., Tuna K.M., Alli A.A., Tribulo P., Hansen P.J., Koh J., Paula-Lopes F.F. Follicular fluid exosomes act on the bovine oocyte to improve oocyte competence to support development and survival to heat shock. Reproduction, Fertility, and Development, 2019, 31(5): 888-897 CrossRef
  26. Singina G.N., Chinarov R.Yu., Lukanina V.A., Voroshbit T.A. The effect of prolactin on the quality of heifer oocytes retrieved by transvaginal puncture of follicles. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2021, 56(6): 1148-1155 CrossRef
  27. Parrish J.J. Bovine in vitro fertilization: in vitro oocyte maturation and sperm capacitation with heparin. Theriogenology, 2014, 81(1): 67-73 CrossRef
  28. Singina G.N. Change of culture medium positively influences the development and quality of in vitro cattle embryos. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2022, 57(6): 1197-1207 CrossRef
  29. Kowal J., Arras G., Colombo M., Jouve M., Morath J.P., Primdal-Bengtson B., Dingli F., Loew D., Tkach M., Théry C. Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proceedings of the National Academy of Sciences of the United States of America, 2016, 113(8): E968-E977 CrossRef
  30. Pavani K.C., Hendrix A., Van Den Broeck W., Couck L., Szymanska K., Lin X., De Koster J., Van Soom A., Leemans B. Isolation and characterization of functionally active extracellular vesicles from culture medium conditioned by bovine embryos in vitro. International Journal of Molecular Sciences, 2018, 20(1): 38 CrossRef
  31. Hung W.-T., Navakanitworakul R., Khan T., Zhang P., Davis J.S., McGinnis L.K., Christenson L.K. Stage-specific follicular extracellular vesicle uptake and regulation of bovine granulosa cell proliferation. Biology of Reproduction, 2017, 97(4): 644-655 CrossRef
  32. da Silveira J.C., Andrade G.M., Del Collado M., Sampaio R.V., Sangalli J.R., Silva L.A., Pinaffi F.V.L., Jardim I.B., Cesar M.C., Nogueira M.F.G., Cesar A.S.M., Coutinho L.L., Pereira R.W., Perecin F., Meirelles F.V. Supplementation with small-extracellular vesicles from ovarian follicular fluid during in vitro production modulates bovine embryo development. PLoS ONE, 2017, 12(6): e0179451 CrossRef
  33. Giacomini E., Makieva S., Murdica V., Vago R., Viganó P. Extracellular vesicles as a potential diagnostic tool in assisted reproduction. Current Opinion in Obstetrics and Gynecology, 2020, 32(3): 179-184 CrossRef
  34. Shedova E.N., Singina G.N., Uzbekova S. Uzbekov R., Lukanina V.A., Tsyndrina E.V. Effect of extracellular vesicles of follicular origin during in vitro maturation and ageing of bovine oocytes on embryo development after in vitro fertilization. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2022, 57(6): 1178-1187 CrossRef
  35. Lopera-Vásquez R., Hamdi M., Fernandez-Fuertes B., Maillo V., Beltrán-Breña P., Calle A., Redruello A., López-Martín S., Gutierrez-Adán A., Yañez-Mó M., Ramirez M.Á., Rizos D. Extracellular vesicles from BOEC in in vitro embryo development and quality. PLoS ONE, 2016, 11(2): e0148083 CrossRef

 

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