Animal Reproduction (AR)
https://animal-reproduction.org/article/doi/10.1590/1984-3143-AR2024-0039
Animal Reproduction (AR)
SHORT COMMUNICATION

First isolation and characterization of caprine oviduct fluid extracellular vesicles

Roberto Mendes Júnior; Agostinho Soares de Alcântara Neto; Gildas Mbemya Tetaping; Marco Aurélio Schiavo Novaes; Vanessa Barbosa Pinheiro Gonçalves; João Xavier da Silva Neto; José Jonathas Albuquerque de Almeida; Maria Izabel Florindo Guedes; Luzia Kalyne Almeida Moreira Leal; Joanna Maria Gonçalves Souza-Fabjan; Roberto Nicolete; Deborah de Melo Magalhães Padilha; José Ricardo de Figueiredo; Ana Paula Ribeiro Rodrigues

Downloads: 0
Views: 20

Abstract

Oviduct fluid extracellular vesicles (oEV) are essential for periconceptional events. The presence of EV has already been identified in the oviduct fluid (OF) from mammalian species, except in caprine. Therefore, this study aimed to isolate and characterize the caprine oEV (coEV). Initially, in Experiment 1, coEV were isolated from the OF of either each animal individually or from a pool of three animals. In experiment 2, coEV were isolated during the follicular or luteal phases of the estrous cycle. The coEV were characterized by size distribution, polydispersity index (PDI), and zeta potential using dynamic light scattering (DLS) analysis, as well as, by transmission electron microscopy (TEM) and dot blotting (DB). Our results indicated that the physicochemical characteristics of the coEV were similar (P > 0.05), regardless of the isolation method (individual or pool). However, coEV collected during the luteal phase were larger (P < 0.05) than those during the follicular phase. The TEM showed spherical and cup-shaped particles, characteristic of exosomes. The DB revealed the presence of exosomal proteins involved in the biogenesis of coEV. In conclusion, it is possible to isolate and characterize coEV from a single caprine female and the estrous cycle phase influences the vesicles average size and PDI.

Keywords

estrous cycle, nanoparticles, oviductosomes, physicochemical characteristics

References

Alcântara-Neto AS, Fernandez-Rufete M, Corbin E, Tsikis G, Uzbekov R, Garanina AS, Coy P, Almiñana C, Mermillod P. Oviduct fluid extracellular vesicles regulate polyspermy during porcine in vitro fertilisation. Reprod Fertil Dev. 2020;32(4):409-18. http://doi.org/10.1071/RD19058. PMid:31775998.

Almiñana C, Bauersachs S. Extracellular vesicles in the oviduct: progress, challenges and implications for the reproductive success. Bioengineering. 2019;6(2):32. http://doi.org/10.3390/bioengineering6020032. PMid:31013857.

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;19(1):622. http://doi.org/10.1186/s12864-018-4982-5. PMid:30134841.

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72(1-2):248-54. http://doi.org/10.1016/0003-2697(76)90527-3. PMid:942051.

Bunggulawa EJ, Wang W, Yin T, Wang N, Durkan C, Wang Y, Wang G. Recent advancements in the use of exosomes as drug delivery systems. J Nanobiotechnology. 2018;16(1):81. http://doi.org/10.1186/s12951-018-0403-9. PMid:30326899.

Danaei M, Dehghankhold M, Ataei S, Hasanzadeh Davarani F, Javanmard R, Dokhani A, Khorasani S, Mozafari MR. Impact of particle size and polydispersity index on the clinical applications of lipidic nanocarrier systems. Pharmaceutics. 2018;10(2):57. http://doi.org/10.3390/pharmaceutics10020057. PMid:29783687.

Huang A, Isobe N, Yoshimura Y. Changes in localization and density of CD63-positive exosome-like substances in the hen oviduct with artificial insemination and their effect on sperm viability. Theriogenology. 2017;101:135-43. http://doi.org/10.1016/j.theriogenology.2017.06.028. PMid:28708510.

Jamaludin NA, Thurston LM, Witek KJ, Meikle A, Basatvat S, Elliott S, Hunt S, Andronowska A, Fazeli A. Efficient isolation, biophysical characterisation and molecular composition of extracellular vesicles secreted by primary and immortalised cells of reproductive origin. Theriogenology. 2019;135:121-37. http://doi.org/10.1016/j.theriogenology.2019.06.002. PMid:31207473.

Javadi M, Rad JS, Pashaiasl M, Farashah MSG, Roshangar L. The effects of plasma-derived extracellular vesicles on cumulus expansion and oocyte maturation in mice. Reprod Biol. 2022;22(1):100593. http://doi.org/10.1016/j.repbio.2021.100593. PMid:34906824.

Kandiel MMM, Watanabe G, Abdel-Ghaffar AE, Sosa GA, Abou-El Roos ME, El-Azab AS, El-Azab I, Li JY, Manabe N, Taya K. Ovarian follicular dynamics and hormonal changes in goats during early pregnancy. J Reprod Dev. 2010;56(5):520-6. http://doi.org/10.1262/jrd.09-179T. PMid:20675962.

Laezer I, Palma-Vera SE, Liu F, Frank M, Trakooljul N, Vernunft A, Schoen J, Chen S. Dynamic profile of EVs in porcine oviductal fluid during the periovulatory period. Reproduction. 2020;159(4):371-82. http://doi.org/10.1530/REP-19-0219. PMid:31990667.

Lennon KM, Wakefield DL, Maddox AL, Brehove MS, Willner AN, Garcia-Mansfield K, Meechoovet B, Reiman R, Hutchins E, Miller MM, Goel A, Pirrotte P, Van Keuren-Jensen K, Jovanovic-Talisman T. Single molecule characterization of individual extracellular vesicles from pancreatic cancer. J Extracell Vesicles. 2019;8(1):1685634. http://doi.org/10.1080/20013078.2019.1685634. PMid:31741725.

Wright A, Snyder OL, Christenson LK, He H, Weiss ML. Effect of pre-processing storage condition of cell culture-conditioned medium on extracellular vesicles derived from human umbilical cord-derived mesenchymal stromal cells. Int J Mol Sci. 2022;23(14):7716. http://doi.org/10.3390/ijms23147716. PMid:35887064.

Yi C, Ni Y, Sun P, Gao T, Li K. Differential size distribution and estrogen receptor cargo of oviductal extracellular vesicles at various stages of estrous cycle in mice. Reprod Sci. 2022;29(10):2847-58. http://doi.org/10.1007/s43032-022-00862-w. PMid:35137347.
 


Submitted date:
02/25/2024

Accepted date:
07/25/2024

67129e25a953951e714d9902 animreprod Articles
Links & Downloads

Anim Reprod

Share this page
Page Sections