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Focal adhesion kinase PTK2 autophosphorylation is not required for the activation of sodium–hydrogen exchange by decreased cell volume in the preimplantation mouse embryo

Published online by Cambridge University Press:  07 June 2019

Jane C. Fenelon
Affiliation:
Ottawa Hospital Research Institute, Ottawa, Ontario, Canada Division of Reproductive Medicine, Department of Obstetrics and Gynaecology, and Department of Cellular and Molecular Medicine, University of Ottawa Faculty of Medicine, Ottawa, Ontario, Canada
Baozeng Xu
Affiliation:
Ottawa Hospital Research Institute, Ottawa, Ontario, Canada Division of Reproductive Medicine, Department of Obstetrics and Gynaecology, and Department of Cellular and Molecular Medicine, University of Ottawa Faculty of Medicine, Ottawa, Ontario, Canada Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, China
Jay M. Baltz*
Affiliation:
Ottawa Hospital Research Institute, Ottawa, Ontario, Canada Division of Reproductive Medicine, Department of Obstetrics and Gynaecology, and Department of Cellular and Molecular Medicine, University of Ottawa Faculty of Medicine, Ottawa, Ontario, Canada
*
*Address for correspondence: Jay M. Baltz. Ottawa Hospital Research Institute, Ottawa, Ontario, Canada. Tel: +1 613 737 8899 ext. 79763. E-mail: [email protected]

Summary

Recovery from decreased cell volume is accomplished by a regulated increase of intracellular osmolarity. The acute response is activation of inorganic ion transport into the cell, the main effector of which is the Na+/H+ exchanger NHE1. NHE1 is rapidly activated by a cell volume decrease in early embryos, but how this occurs is incompletely understood. Elucidating cell volume-regulatory mechanisms in early embryos is important, as it has been shown that their dysregulation results in preimplantation developmental arrest. The kinase JAK2 has a role in volume-mediated NHE1 activation in at least some cells, including 2-cell stage mouse embryos. However, while 2-cell embryos show partial inhibition of NHE1 when JAK2 activity is blocked, NHE1 activation in 1-cell embryos is JAK2-independent, implying a requirement for additional signalling mechanisms. As focal adhesion kinase (FAK aka PTK2) becomes phosphorylated and activated in some cell types in response to decreased cell volume, we sought to determine whether it was involved in NHE1 activation in the early mouse embryo. FAK activity requires initial autophosphorylation of a tyrosine residue, Y397. However, FAK Y397 phosphorylation levels were not increased in either 1- or 2-cell embryos after cell volume was decreased. Furthermore, the selective FAK inhibitor PF-562271 did not affect NHE1 activation at concentrations that essentially eliminated Y397 phosphorylation. Thus, autophosphorylation of FAK Y397 does not appear to be required for NHE1 activation induced by a decrease in cell volume in early mouse embryos.

Type
Research Article
Copyright
© Cambridge University Press 2019 

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