Competence to induced partenogenesis of Bos Taurus oocytes, depending on the gamete functional status and its origin (follicle diameter)
https://doi.org/10.31043/2410-2733-2020-1-15-21
Abstract
The development of effective methods for obtaining parthenogenetic embryos is currently being considered in a new perspective as a promising technology for creating homozygous lines of embryonic stem cells, genetically identical animals. Parthenogenetic embryos of animals of different species are used as a model for studying imprinting mechanisms. In the present study, Bos Taurus parthenogenetic embryos were obtained by cold shock induction (exposure for 20 minutes at a temperature of -0 - (-4o) C) of after oocytes culture in TC-119 medium with 10% fetal bovine serum, 50 ng / ml prolactin, 50 μg / ml of gentamicin together with 1 × 106 granulosa cells per ml of medium for 24 hours. Previously, the initial population of donor oocytes from the ovaries was evaluated with BCB test to assess functional status. For this, the oocyte-cumulus complexes were placed in a solution of 13 Mm brilliant crystalline blue - BCB (B-5388, Sigma) for 60 minutes at a temperature of 38.50 C in an atmosphere of 90% humidity. After the expiration of the BCB exposure time, the oocytes were ranked as follows: oocytes with a blue cytoplasm - BCB + oocytes (oocytes that completed their growth phase) and oocytes with unpainted ooplasm - BCB- oocytes (growing oocytes). It was found that the functional status (growing oocytes or oocytes that have finished growth phase) of oocytes and their origin (the diameter of the follicles from which the gametes were isolated) determine their competence for induced parthenogenesis. Oocytes that completed the growth phase have high potentials to cold shock-induced parthenogenesis, which is expressed in a significant increase in the proportion of embryos developed from them at all stages of preimplantation development (percentage of cleavage: 87% vs 11%, P <0.001; level of late morula, blastocysts: 32 % vs 6%, P <0.001). Monitoring of competency indicators to parthenogenesis of Bos Taurus oocytes isolated from follicles of different diameters (<3 mm, 3 mm -5 mm,> 5 mm-8 mm) revealed low competence to cold shock-induced parthenogenesis of oocytes isolated from follicles less than 3 mm in diameter ( the percentage of cleavage: 20% vs 67% and 87%, P <0.001, respectively; level of late morula, blastocysts: 9% vs 22% and 37%, P <0.001, respectively).
About the Authors
T. Kuzmina
Russian research institute of farm animal genetics and breeding — branch of the L. K. Ernst Federal science center for animal husbandry
Russian Federation
Russian Federation
Dr. Habil. (Biol. Sci.)
St. Petersburg, Pushkin, Moskovskoe shosse, 55a, 196601
H. Alm
Leibniz Institute for Farm Animal Biology (FBN)
Germany
Germany
Alm Hanna — PhD (Biology)
Wilhelm-Stahl-Allee 2, 18196, Dummerstorf
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Review
For citations:
Kuzmina T., Alm H. Competence to induced partenogenesis of Bos Taurus oocytes, depending on the gamete functional status and its origin (follicle diameter). Genetics and breeding of animals. 2020;(1):15-21. (In Russ.) https://doi.org/10.31043/2410-2733-2020-1-15-21
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