Inhibition of Na+, K+ -ATPase with ouabain is detrimental to equine blastocysts
Anim. Reprod. (Online)
; 17(1): e20190079, 2020. ilus
Article
em En
| VETINDEX
| ID: biblio-1461483
Biblioteca responsável:
BR68.1
Localização: BR68.1
ABSTRACT
Although equine blastocysts ≤ 300 µm in diameter can be successfully vitrified, larger equine blastocysts are not good candidates for cryopreservation. As Na+, K+-ATPase is involved in maintaining blastocyst expansion, perhaps inhibition of this enzyme would be a viable method of reducing blastocyst diameter prior to cryopreservation. Objectives were to evaluate effects of ouabain-induced inhibition of Na+, K+-ATPase in equine blastocysts. Sixteen mares were ultrasonographically monitored, given deslorelin acetate to induce ovulation, and inseminated. Embryos (D7 and D9) were harvested and Na+, K+-ATPase inhibited for 1 or 6 h by exposure to 10-6 M ouabain, either natural ouabain or conjugated to fluorescein (OuabainFL), during incubation at 37° C. Evaluations included morphometric characteristics (bright field microscopy) and viability (Hoescht 33342 + propidium iodide). Blastocysts incubated for 6 h in Holding medium + ouabain (n=3) had, on average, a 45.7% reduction in diameter, with adverse morphologic features and no re-expansion after subsequent incubation in Holding medium for 12 h. In subsequent studies, even a 1-h exposure to Ouabain or OuabainFL, caused similar reductions, namely 38.7 ± 6.7% (n=5) and 33.6 ± 3.3% (n=7) for D7 and D9 blastocysts, respectively. Ouabain binding was confirmed after OuabainFL exposition and all embryos (n=12) lost viability. We concluded that Na+, K+-ATPase inhibition with ouabain caused death of equine blastocysts and therefore was not a viable method of reducing blastocyst size prior to cryopreservation.
Palavras-chave
Texto completo:
1
Base de dados:
VETINDEX
Assunto principal:
Ouabaína
/
Blastocisto
/
ATPase Trocadora de Sódio-Potássio
/
Cavalos
Limite:
Animals
Idioma:
En
Revista:
Anim. Reprod.
/
Anim. Reprod. (Online)
Ano de publicação:
2020
Tipo de documento:
Article