Morphological evaluation of Prochilodus lineatus embryos after vitrification-thawing in high-osmolarity cryoprotectant solution.

Autor: Costa RDS; Department of Biology and Animal Science, Universidade Estadual Paulista 'Júlio de Mesquita Filho', Ilha Solteira, Brazil.; Pós-Graduação em Biologia Animal, Universidade Estadual Paulista 'Júlio de Mesquita Filho', São José do Rio Preto, Brazil., Capuzzo CS; Department of Biology and Animal Science, Universidade Estadual Paulista 'Júlio de Mesquita Filho', Ilha Solteira, Brazil., Ribeiro CDS; Department of Biology and Animal Science, Universidade Estadual Paulista 'Júlio de Mesquita Filho', Ilha Solteira, Brazil., Verissimo-Silveira R; Department of Biology and Animal Science, Universidade Estadual Paulista 'Júlio de Mesquita Filho', Ilha Solteira, Brazil., Siqueira-Silva DH; Instituto de Estudos em Saúde e Biológicas - IESB, Universidade Federal do Sul e Sudeste do Pará, Marabá, Brazil., Senhorini JA; National Center for Research and Conservation of Continental Fish at the Chico Mendes Institute of Biodiversity Conservation - CEPTA/ICMBio, Pirassununga, São Paulo, Brazil., Ninhaus-Silveira A; Pós-Graduação em Biologia Animal, Universidade Estadual Paulista 'Júlio de Mesquita Filho', São José do Rio Preto, Brazil.
Jazyk: angličtina
Zdroj: Reproduction in domestic animals = Zuchthygiene [Reprod Domest Anim] 2018 Dec; Vol. 53 (6), pp. 1353-1358. Date of Electronic Publication: 2018 Jul 16.
DOI: 10.1111/rda.13249
Abstrakt: We aimed to vitrify embryos of Prochilodus lineatus in a high-osmolarity cryoprotectant solution, evaluating, after the vitrification-thawing process, their morphological changes. Thus, 240 embryos in the 20-somite phase (20S) were exposed for 20 min to one main internal cryoprotectant solution (1,2-propanediol-PROP), divided into four immersion sequence steps of five minutes each. The first three steps were performed in solutions containing only a main internal cryoprotectant (PROP-2, 3 and 4 M), and the fourth step in a high-osmolarity solution combining internal (PROP + dimethyl sulphoxide-Me 2 SO) and external cryoprotectants (sucrose-SUC). The final concentration of vitrification was PROP 5 M + Me 2 SO 5 M + SUC 0.2 M. During vitrification, the straws exhibited a translucent solid appearance; however, during thawing, their structure became totally opaque and white. After thawing, the embryos suffered an increase in volume and presented morphological changes including protrusions on the surface of the yolk sac, yolk sac rupture, and optical vesicle degradation. On the inside, we observed intercellular spaces and a yolk syncytial layer (YSL) with altered chromatin. Yet, structures such as somites, neural tube, endoderm and epidermis presented cells with a nucleus and integral mitochondria. We conclude that the use of the tested cryoprotectant solution permits the formation of a vitreous solid and preserves part of the cells of the blastoderm. Yet, the heating protocol does not control recrystallization, resulting in the formation of serious morphological anomalies that prevent the preservation of the embryonic unit.
(© 2018 Blackwell Verlag GmbH.)
Databáze: MEDLINE
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