Vitrification protocol for immature Brycon orbignyanus ovarian tissue as an extinction escape strategy.

Autor: Santos Marques L; Animal Science Research Program of Federal University of Rio Grande Do Sul, Bento Gonçalves, 7712, 91540-000, Porto Alegre, RS, Brazil., Rodrigues de Freitas T; Animal Science Research Program of Federal University of Rio Grande Do Sul, Bento Gonçalves, 7712, 91540-000, Porto Alegre, RS, Brazil., Batista Rodrigues R; Animal Science Research Program of Federal University of Rio Grande Do Sul, Bento Gonçalves, 7712, 91540-000, Porto Alegre, RS, Brazil., Dos Santos Teixeira N; Animal Science Research Program of Federal University of Rio Grande Do Sul, Bento Gonçalves, 7712, 91540-000, Porto Alegre, RS, Brazil., Pérez-Atehortúa M; Animal Science Research Program of Federal University of Rio Grande Do Sul, Bento Gonçalves, 7712, 91540-000, Porto Alegre, RS, Brazil., Rosa-Silva HT; Postgraduate Program in Biological Sciences: Biochemistry, Department of Biochemistry of Federal University of Rio Grande Do Sul, Ramiro Barcelos, 2600, 90035-003, Porto Alegre, RS, Brazil., Fonseca Moreira JC; Postgraduate Program in Biological Sciences: Biochemistry, Department of Biochemistry of Federal University of Rio Grande Do Sul, Ramiro Barcelos, 2600, 90035-003, Porto Alegre, RS, Brazil., Streit DP Jr; Animal Science Research Program of Federal University of Rio Grande Do Sul, Bento Gonçalves, 7712, 91540-000, Porto Alegre, RS, Brazil. Electronic address: danilo.streit@ufrgs.br.
Jazyk: angličtina
Zdroj: Cryobiology [Cryobiology] 2021 Dec; Vol. 103, pp. 116-122. Date of Electronic Publication: 2021 Aug 28.
DOI: 10.1016/j.cryobiol.2021.08.004
Abstrakt: Piracanjunba (Brycon orbignyanus) is an endangered South American fish, and ovarian tissue cryopreservation is an alternative method for preserving maternal germplasm and genetic diversity. Therefore, our aim was to test a vitrification protocol for ovarian tissue containing primary growth (PG) oocytes of B. orbignyanus as a strategy to avoid the threat of extinction. Two vitrification solutions were evaluated (VS1: 1.5 M methanol + 4.5 M propylene glycol and VS2: 1.5 M methanol + 5.5 M Me 2 SO) and compared using control/fresh ovarian tissue. After vitrification, the following factors were analyzed: membrane integrity using trypan blue, morphology using a histological assessment, oxidative stress (total reactive antioxidant potential (TRAP) and reduced thiol [-SH]), mitochondrial activity using MTT, and DNA damage using a comet assay. The vitrified oocytes (VS1 = 24.3 ± 0.49% and VS2 = 24.8 ± 0.69%) showed higher DNA damage than the control group (control = 20.7 ± 1.03%) (P = 0.004). In contrast, in most evaluations (membrane integrity, membrane damage, oxidative stress, and mitochondrial activity), there were no discernible differences between the control group and the vitrified samples. In addition, oocyte (P = 0.883) and nuclear diameter (P = 0.118) did not change after vitrification. VS2 treatment resulted in higher nuclear damage (15.7 ± 1.45%) than in the control treatment (3.5 ± 1.19%); however, VS1 treatment did not result in significantly more damage (9.5 ± 3.01%) than in the control (P = 0.015). Therefore, the protocol for ovarian tissue vitrification tested in this study resulted in high maintenance of PG oocyte cell integrity, making it a promising alternative for B. orbignyanus maternal genome preservation.
(Copyright © 2021. Published by Elsevier Inc.)
Databáze: MEDLINE
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