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Hum. Reprod. Advance Access published online on March 3, 2009
Human Reproduction, doi:10.1093/humrep/dep010
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© The Author 2009. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Mitogen-activated protein kinase (MAPK) pathways mediate embryonic responses to culture medium osmolarity by regulating Aquaporin 3 and 9 expression and localization, as well as embryonic apoptosis

Christine E. Bell1,2,3, Nathalie M.K. Larivière1,2,3,4, Patricia H. Watson2,4,5 and Andrew J. Watson1,2,3,4,5,6

1 Department of Obstetrics and Gynaecology, The University of Western Ontario, London, Ontario, Canada 2 Department of Physiology and Pharmacology, The University of Western Ontario, London, Ontario, Canada 3 Children's Health Research Institute, London, Ontario, Canada 4 Lawson Health Research Institute, London, Ontario, Canada 5 Department of Medicine, The University of Western Ontario, London, Ontario, Canada

6 Correspondence address. E-mail: awatson{at}uwo.ca

BACKGROUND: In order to advance the development of culture conditions and increase the potential for supporting normal preimplantation embryo development in vitro, it is critical to define the mechanisms that early embryos utilize to survive in culture. We investigated the mechanisms that embryos employ in response to culture medium osmolarity. We hypothesized that mitogen-activated protein kinase (MAPK) pathways mediate responses to hyperosmotic stress by regulating Aquaporin (AQP) 3 and 9 expression as well as embryonic apoptosis.

METHODS: Real-time reverse transcription and polymerase chain reaction and whole-mount immunofluorescence were used to determine the relative mRNA levels and protein localization patterns of AQP 3 and 9 after hyperosmotic medium treatment.

RESULTS: At 6 and 24 h, a significant increase in Aqp 3 and 9 mRNA was observed in the sucrose hyperosmotic treatment compared with standard medium and glycerol controls. Blockade of MAPK14/11 negated the increase in Aqp 3 and 9 mRNA levels, whereas culture in a MAPK8 blocker did not. Hyperosmotic sucrose treatment significantly increased embryonic apoptosis which was negated in the presence of MAPK8 blocker, but not MAPK14/11 blocker.

CONCLUSIONS: MAPK14/11 activation is a component of the rapid adaptive stress response mechanism that includes the effects of AQP mRNA expression and protein localization, whereas the MAPK8 pathway is a regulator of apoptosis.

Key words: programmed cell death/water channels/blastocyst/embryo culture

Submitted on June 18, 2008; resubmitted on December 19, 2008; accepted on January 7, 2009.


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