Artificial blastocyst collapse prior to vitrification significantly improves Na + /K + -ATPase-dependent post-warming blastocoel re-expansion kinetics without inducing endoplasmic reticulum stress gene expression in the mouse

L. A. Frank, R. D. Rose, M. R. Anastasi, T. C.Y. Tan, M. F. Barry, J. G. Thompson, H. M. Brown

Research output: Contribution to journalArticle

2 Citations (Scopus)


Blastocoel expansion during embryo development is known to be reliant on the Na + /K + -ATPase pump, but little is known about the relative contribution of active (Na + /K + -ATPase pump) and facilitated diffusion (aquaporins) water transport during blastocoel re-expansion after vitrification. The aims of this study were to examine potential effects of artificial blastocoel collapse (ABC) on markers of embryo stress and the contribution of active and facilitated diffusion water transport mechanisms to blastocoel re-expansion. Day 5 mouse embryos were vitrified using either a standard protocol, laser pulse ABC, a hyperosmotic sucrose ABC protocol or both laser pulse and sucrose. Using real-time polymerase chain reaction, no differences were found in the gene expression of the endoplasmic reticulum (ER) stress markers activating transcription factor 4 (Atf4) or heat shock protein 90-alpha (Hsp90α) 2 h after warming. Similarly, expression of the Na + /K + -ATPase pump gene, ATPase, Na+/K+ transporting, beta 1 polypeptide (Atp1b1) and protein did not differ between groups. Aquaporin 8 (Aqp8) gene expression was significantly lower in the laser + sucrose ABC group than in fresh controls, and aquaporin 3 (Aqp3) expression significantly higher in standard vitrified embryos compared with all other groups. Ouabain, a potent and specific Na + /K + -ATPase pump inhibitor, inhibited blastocoel re-expansion in both standard protocol- and laser ABC-vitrified embryos, reducing both groups to the same rate of re-expansion 3 h after warming. These results demonstrate that ABC before vitrification does not alter mRNA or protein expression of Na + /K + -ATPase, or mRNA levels of ER stress genes Atf4 and Hsp90α. Activity of the pump may be increased in ABC embryos, with potential compensation by AQP3 when it is compromised.

Original languageEnglish
Pages (from-to)294-305
Number of pages12
JournalReproduction, Fertility and Development
Issue number2
Publication statusPublished - 1 Jan 2019
Externally publishedYes


  • assisted blastocyst collapse
  • embryo collapse
  • embryo re-expansion kinetics
  • laser-assisted collapse

ASJC Scopus subject areas

  • Biotechnology
  • Reproductive Medicine
  • Animal Science and Zoology
  • Molecular Biology
  • Genetics
  • Endocrinology
  • Developmental Biology

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