Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 2017, V. 52, № 4, pp. 776-784
(how to cite this article)

doi: 10.15389/agrobiology.2017.4.776eng

UDC 636.2:591.39

Acknowledgements:
Supported by grant from Russian Science Foundation (project 16-16-10069)

 

BOVINE OOCYTE ABILITY TO EMBRYONIC DEVELOPMENT WHEN
MATURING IN DIFFERENT TWO-PHASE CULTURE SYSTEMS

G.N. Singina, I.Yu. Lebedeva, E.N. Shedova, T.E. Taradajnic,
O.S. Mityashova, E.V. Tsyndrina, S.S. Danch

L.K. Ernst Federal Science Center for Animal Husbandry, Federal Agency of Scientific Organizations, 60, pos. Dubrovitsy, Podolsk District, Moscow Province, 142132 Russia,
e-mail g_singina@mail.ru (corresponding author), irledv@mail.ru, shedvek@yandex.ru,almaatinka25@rambler.ru, mityashova_o@mail.ru, danch.s.s@mail.ru, vip.krilochkina@mail.ru

The authors declare no conflict of interests

ORCID:

Singina G.N.
orcid.org/0000-0003-0198-9757

Mityashova O.S.
orcid.org/0000-0002-0401-5088

Lebedeva I.Yu.
orcid.org/0000-0002-7815-7900

Tsyndrina E.V.
orcid.org/0000-0002-3263-2358

Shedova E.N.
orcid.org/0000-0002-9642-2384

Danch S.S.
orcid.org/0000-0002-0198-2013

Taradajnic T.E.
orcid.org/0000-0002-1139-6667

 

Received May 16, 2017

 

Existing approaches to a two-phase method of culture of cattle oocytes do not take into account their specific demands during maturation from metaphase I to metaphase II including the need for normalization of profiles of sex steroid hormones in a culture medium. The aim of the presented research was to compare the developmental competence of bovine (Bos taurus taurus) oocytes matured in conventional single-phase and different two-phase systems. We have studied for the first time the ovum ability to develop to the blastocyst stage and the quality of embryos produced when replacing the standard medium at the second stage of culture with a medium free of follicle-stimulating hormone (FSH) as well as by a granulosa cell culture. When using the single-phase system, cumulus-oocyte complexes (COCs) were cultured for 24 h in the medium TCM 199 containing 10 % fetal bovine serum (FBS), 10 µg/ml FSH, and 10 µg/ml luteinizing hormone (LH). In the two-phase system, oocytes matured in the same conditions for first 12 h. Then COCs were transferred to a new medium (TCM 199 containing 10% FBS or the same medium supplemented with 5 µg/ml LH) and cultured for 12 h in the presence or in the absence of granulosa cells. After 24 h of maturation, oocyte fertilization was performed. In embryos that developed to the stages of late morula and blastocyst on day 7 of culture, the degree of apoptosis and the total number of nuclei were assessed (by the TUNEL method and DAPI staining, respectively). In media collected after oocyte maturation, levels of progesterone and 17β-estradiol were determined by enzyme immunoassay. The proportions of oocytes entered the first cleavage division (57.6-68.1 %) or developed to the stages of late morula/blastocyst (16.7-20.7 %) were not associated with the culture method or medium. The transfer of oocytes after 12 h of maturation to the medium free of gonadotropic hormones resulted in 1.3-1.4-fold increase in the total number of nuclei in the late morulae/blastocysts both in the absence and in the presence of granulosa cells, whereas this number decreased to the level of single-phase control on addition of LH. At the same time the proportion of apoptotic nuclei (2.3-6.9 %) did not depend on the system of oocyte maturation or the effect of LH. In the two-phase system, the level of 17b-estradiol in the maturation medium (with and without LH) was 1.3-1.4-fold lower (p < 0.01) as compared to the single-phase system. On the contrary, co-culture of COCs with granulosa cells led to a rise in the concentration of 17b-estradiol in the culture medium containing LH (from 418±16 to 496±26 pg/ml, p < 0.05). Meanwhile, no differences in the progesterone level in the culture medium of COCs of oocytes after 12 h of culture to the medium free of gonadotropic hormones causes an increase in the quality of the late morulae/blastocysts produced as well as leads to normalization of the 17b-estradiol profile in the culture medium at the terminal step of the ovum maturation.

Keywords: bovine oocytes, two-phase system of in vitro maturation, embryonic development, sex steroid hormones.

 

Full article (Rus)

Full text (Eng)

 

REFERENCES

  1. Mapletoft R.J, Hasler J.F. Assisted reproductive technologies in cattle: a review. Rev. Sci. Tech., 2005, 24(1): 393-403
  2. Lonergan P., Fair T. In vitro-produced bovine embryos: dealing with the warts. Theriogenology, 2008, 69: 17-22 CrossRef
  3. Sanchez F., Smitz J. Molecular control of oogenesis. Biochim. Biophys. Acta, 2012, 1822: 1896-1912 CrossRef
  4. Blondin P., Coenen K., Guilbault L.A., Sirard M.A. In vitro production of bovine embryos: Developmental competence is acquired before maturation. Theriogenology, 1997, 47: 1061-1075 CrossRef
  5. Eppig J.J., Pendola F.L., Wigglesworth K., Pendola J.K. Mouse oocytes regulate metabolic cooperativity between granulosa cells and oocytes: amino acid transport. Biol. Reprod., 2005, 73: 351-357 CrossRef
  6. Thompson J.G., Lane M., Gilchrist R.B. Metabolism of the bovine cumulus-oocyte complex and influence on subsequent developmental competence. Soc. Reprod. Fertil. Suppl., 2007, 64: 179-190.
  7. Gilchrist R.B., Thompson J.G. Oocyte maturation: emerging concepts and technologies to improve developmental potential in vitro. Theriogenology, 2007, 67: 6-15 CrossRef
  8. Hussein T.S., Sutton-McDowall M.L., Gilchrist R.B., Thompson J.G. Temporal effects of exogenous oocyte-secreted factors on bovine oocyte developmental competence during IVM. Reprod. Fertil. Dev., 2011, 23: 576-584 CrossRef
  9. Sirard M.A. Resumption of meiosis: mechanism involved in meiotic progression and its relation with developmental competence. Theriogenology, 2001, 55: 1241-1254 CrossRef
  10. Bilodeau-Goeseels S. Bovine oocyte meiotic inhibition before in vitro maturation and its value to in vitro embryo production: does it improve developmental competence? Reprod. Domest. Anim., 2011, 47: 687-693 CrossRef
  11. Zhang J., Wei Q., Cai J., Zhao X., Ma B. Effect of C-type natriuretic peptide on maturation and developmental competence of goat oocytes matured in vitro. PLoS ONE, 2015, 10(7): e0132318 CrossRef
  12. Franciosi F., Coticchio G., Lodde V., Tessaro I., Silvia C. Modina S.C., Fadini R., Dal Canto M., Renzini M.M., Albertini D.F., Luciano A.M. Natriuretic peptide precursor C delays meiotic resumption and sustains gap junction-mediated communication in bovine cumulus-enclosed oocytes. Biol. Reprod., 2014, 91(3): 61 CrossRef
  13. Oliveira e Silva I., Vasconcelos R.B., Caetano J.V., Gulart L.V., Camargo L.S., Báo S.N., Rosa e Silva A.A. Induction of reversible meiosis arrest of bovine oocytes using a two-step procedure under defined and nondefined conditions. Theriogenology, 2011, 75(6): 1115-1124 CrossRef
  14. Guemra S., da Silva Santo E., Zanin R., Monzani P.S., Sovernigo T.C., Ohashi O.M., Verde Leal C.L., Adona P.R. Effect of temporary meiosis block during prematuration of bovine cumulus-oocyte complexes on pregnancy rates in a commercial setting for in vitro embryo production. Theriogenology, 2014, 81(7): 982-987 CrossRef
  15. Pavlok A., Lapathitis G., Cech S., Kubelka M., Lopatárová M., Holý L., Klíma J., Motlík J., Havlícek V. Simulation of intrafollicular conditions prevents GVBD in bovine oocytes: a better alternative to affect their developmental capacity after two-step culture. Mol. Reprod. Dev., 2005, 71(2): 197-208 CrossRef
  16. Mingoti G.Z., Garcia J.M., Rosa-e-Silva A.A. Steroidogenesis in cumulus cells of bovine cumulus-oocyte-complexes matured in vitro with BSA and different concentrations of steroids. Anim. Reprod. Sci., 2002, 69: 175-186 CrossRef
  17. Schoenfelder M., Schams D., Einspanier R. Steroidogenesis during in vitro maturation of bovine cumulus oocyte complexes and possible effects of tri-butyltin on granulosa cells. J. Steroid. Biochem. Mol. Biol., 2003, 84: 291-300 CrossRef
  18. Tosca L., Uzbekova S., Chabrolle C., Dupont J. Possible role of 5´AMP-activated protein kinase in the metformin-mediated arrest of bovine oocytes at the germinal vesicle stage during in vitro maturation. Biol. Reprod., 2007, 77: 452-465 CrossRef
  19. Blaschka C., Wrenzycki C. Temporal pattern of steroid hormone concentrations during in vitro maturation of bovine oocytes. Anim. Reprod., 2015, 12(3): 729.
  20. Gilchrist R.B., Ritter L.J., Armstrong D.T. Oocyte-somatic cell interactions during follicle development in mammals. Anim. Reprod. Sci., 2004, 82-83: 431-446 CrossRef
  21. Singina G.N., Bagirov V.A., Danch S.S., Taradainik T.E., Dotsev A.V., Zinov'eva N.A. In vitro production of interspecies hybrid embryos of cattle (Bos taurus) and wisent (Bison bonasus). Sel’skokhozyaistvennaya biology [Agricultural Biology], 2016, 51(6): 824-829 CrossRef
  22. Parrish J.J., Susko-Parrish J.L., Leibfried-Rutledge M.L., Critser E.S., Eyestone W.H., First N.L. Bovine in vitro fertilization with frozen-thawed semen. Theriogenology, 1986, 25(4): 591-600 CrossRef
  23. Rosenkrans C.F. Jr., First N.L. Effect of free amino acids and vitamins on cleavage and developmental rate of bovine zygotes in vitro. J. Anim. Sci., 1994, 72(2): 434-437 CrossRef
  24. Funahashi H., Day B.N. Effects of the duration of exposure to hormone supplements on cytoplasmic maturation of pig oocytes in vitro. J. Reprod. Fertil., 1993, 98: 179-185 CrossRef
  25. Wang H.F., Isobe N., Kumamoto K., Yamashiro H., Yamashita Y., Terada T. Studies of the role of steroid hormone in the regulation of oocyte maturation in cattle. Reprod. Biol. Endocrinol., 2006, 4: 4 CrossRef

back