Skip to main content Accessibility help

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

  • Jane C. Fenelon (a1) (a2), Baozeng Xu (a1) (a2) (a3) and Jay M. Baltz (a1) (a2)


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.


Corresponding author

*Address for correspondence: Jay M. Baltz. Ottawa Hospital Research Institute, Ottawa, Ontario, Canada. Tel: +1 613 737 8899 ext. 79763. E-mail:


Hide All
Alexander, RT and Grinstein, S (2006) Na+/H+ exchangers and the regulation of volume. Acta Physiol (Oxf) 187, 159167.
Arold, ST (2011) How focal adhesion kinase achieves regulation by linking ligand binding, localization and action. Curr Opin Struct Biol 21, 808813.
Aronson, PS, Nee, J and Suhm, MA (1982) Modifier role of internal H+ in activating the Na+-H+ exchanger in renal microvillus membrane vesicles. Nature 299, 161163.
Arroyo, JP, Kahle, KT and Gamba, G (2013) The SLC12 family of electroneutral cation-coupled chloride cotransporters. Mol Aspects Med 34, 288298.
Baltz, JM and Phillips, KP (1999) Intracellular ion measurements in single eggs and embryos using ion-sensitive fluorophores. In A Comparative Methods Approach to the Study of Oocytes and Embryos, ed. JD Richter, Oxford University Press, New York and Oxford, pp. 3982.
Baltz, JM and Tartia, AP (2010) Cell volume regulation in oocytes and early embryos: connecting physiology to successful culture media. Hum Reprod Update 16, 166176.
Baltz, JM and Zhou, C (2012) Cell volume regulation in mammalian oocytes and preimplantation embryos. Mol Reprod Dev 79, 821831.
Dawson, KM and Baltz, JM (1997) Organic osmolytes and embryos: substrates of the Gly and beta transport systems protect mouse zygotes against the effects of raised osmolarity. Biol Reprod 56, 15501558.
Donowitz, M, Ming Tse, C and Fuster, D (2013) SLC9/NHE gene family, a plasma membrane and organellar family of Na+/H+ exchangers. Mol Aspects Med 34, 236251.
Garnovskaya, MN, Mukhin, YV, Vlasova, TM and Raymond, JR (2003) Hypertonicity activates Na+/H+ exchange through Janus kinase 2 and calmodulin. J Biol Chem 278, 1690816915.
Gatsios, P, Terstegen, L, Schliess, F, Häussinger, D, Kerr, IM, Heinrich, PC and Graeve, L (1998) Activation of the Janus kinase/signal transducer and activator of transcription pathway by osmotic shock. J Biol Chem 273, 2296222968.
Gibb, CA, Poronnik, P, Day, ML and Cook, DI (1997) Control of cytosolic pH in two-cell mouse embryos: roles of H+-lactate cotransport and Na+/H+ exchange. Am J Physiol 273, C404C419.
Hadi, T, Hammer, MA, Algire, C, Richards, T and Baltz, JM (2005) Similar effects of osmolarity, glucose, and phosphate on cleavage past the 2-cell stage in mouse embryos from outbred and F1 hybrid females. Biol Reprod 72, 179187.
Harding, EA, Gibb, CA, Johnson, MH, Cook, DI and Day, ML (2002) Developmental changes in the management of acid loads during preimplantation mouse development. Biol Reprod 67, 14191429.
Hoffmann, EK and Pedersen, SF (2011) Cell volume homeostatic mechanisms: effectors and signalling pathways. Acta Physiol 202, 465485.
Hoffmann, EK, Lambert, IH and Pedersen, SF (2009) Physiology of cell volume regulation in vertebrates. Physiol Rev 89, 193277.
Humphreys, BD, Jiang, L, Chernova, MN and Alper, SL (1995) Hypertonic activation of AE2 anion exchanger in Xenopus oocytes via NHE-mediated intracellular alkalinization. Am J Physiol 268, C201c209.
Jiang, L, Chernova, MN and Alper, SL (1997) Secondary regulatory volume increase conferred on Xenopus oocytes by expression of AE2 anion exchanger. Am J Physiol 272, C191202.
Karaman, MW, Herrgard, S, Treiber, DK, Gallant, P, Atteridge, CE, Campbell, BT, Chan, KW, Ciceri, P, Davis, MI, Edeen, PT, Faraoni, R, Floyd, M, Hunt, JP, Lockhart, DJ, Milanov, ZV, Morrison, MJ, Pallares, G, Patel, HK, Pritchard, S, Wodicka, LM and Zarrinkar, PP (2008) A quantitative analysis of kinase inhibitor selectivity. Nat Biotechnol 26, 127132.
Krump, E, Nikitas, K and Grinstein, S (1997) Induction of tyrosine phosphorylation and Na+/H+ exchanger activation during shrinkage of human neutrophils. J Biol Chem 272, 1730317311.
Lawitts, JA and Biggers, JD (1992) Joint effects of sodium chloride, glutamine, and glucose in mouse preimplantation embryo culture media. Mol Reprod Dev 31, 189194.
Lawitts, JA and Biggers, JD (1993) Culture of preimplantation embryos. Methods Enzymol 225, 153164.
Lunn, JA and Rozengurt, E (2004) Hyperosmotic stress induces rapid focal adhesion kinase phosphorylation at tyrosines 397 and 577: role of Src family kinases and Rho family GTPases. J Biol Chem 279, 4526645278.
Lunn, JA, Jacamo, R and Rozengurt, E (2007) Preferential phosphorylation of focal adhesion kinase tyrosine 861 is critical for mediating an anti-apoptotic response to hyperosmotic stress. J Biol Chem 282, 1037010379.
Malo, ME and Fliegel, L (2006) Physiological role and regulation of the Na+/H+ exchanger. Can J Physiol Pharmacol 84, 10811095.
McGinnis, LK and Kinsey, WH (2015) Role of focal adhesion kinase in oocyte–follicle communication. Mol Reprod Dev 82, 90102.
McGinnis, LK, Carroll, DJ and Kinsey, WH (2011) Protein tyrosine kinase signaling during oocyte maturation and fertilization. Mol Reprod Dev 78, 831845.
McGinnis, LK, Luo, J and Kinsey, WH (2013) Protein tyrosine kinase signaling in the mouse oocyte cortex during sperm–egg interactions and anaphase resumption. Mol Reprod Dev 80, 260272.
Rasmussen, LJ, Muller, HS, Jorgensen, B, Pedersen, SF and Hoffmann, EK (2015) Osmotic shrinkage elicits FAK- and Src phosphorylation and Src-dependent NKCC1 activation in NIH3T3 cells. Am J Physiol Cell Physiol 308, C101c110.
Roberts, WG, Ung, E, Whalen, P, Cooper, B, Hulford, C, Autry, C, Richter, D, Emerson, E, Lin, J, Kath, J, Coleman, K, Yao, L, Martinez-Alsina, L, Lorenzen, M, Berliner, M, Luzzio, M, Patel, N, Schmitt, E, LaGreca, S, Jani, J, Wessel, M, Marr, E, Griffor, M and Vajdos, F (2008) Antitumor activity and pharmacology of a selective focal adhesion kinase inhibitor, PF-562,271. Cancer Res 68, 19351944.
Romero, MF, Chen, AP, Parker, MD and Boron, WF (2013) The SLC4 family of bicarbonate (HCO3 ) transporters. Mol Aspects Med 34, 159182.
Sardet, C, Counillon, L, Franchi, A and Pouyssegur, J (1990) Growth factors induce phosphorylation of the Na+/H+ antiporter, glycoprotein of 110 kD. Science 247, 723726.
Schaller, MD, Hildebr, JD, Shannon, JD, Fox, JW, Vines, RR and Parsons, JT (1994) Autophosphorylation of the focal adhesion kinase, pp125FAK, directs SH2-dependent binding of pp60src. Mol Cell Biol 14, 16801688.
Schlaepfer, DD and Hunter, T (1996) Evidence for in vivo phosphorylation of the Grb2 SH2-domain binding site on focal adhesion kinase by Src-family protein-tyrosine kinases. Mol Cell Biol 16, 56235633.
Schlaepfer, DD, Mitra, SK and Ilic, D (2004) Control of motile and invasive cell phenotypes by focal adhesion kinase. Biochim Biophys Acta Mol Cell Biol Lipids 1692, 77102.
Siyanov, V and Baltz, JM (2013) NHE1 is the sodium–hydrogen exchanger active in acute intracellular pH regulation in preimplantation mouse embryos. Biol Reprod 88, 157.
Steeves, CL, Lane, M, Bavister, BD, Phillips, KP and Baltz, JM (2001) Differences in intracellular pH regulation by Na+/H+ antiporter among 2-cell mouse embryos derived from females of different strains. Biol Reprod 65, 1422.
Thomas, JA, Buchsbaum, RN, Zimniak, A and Racker, E (1979) Intracellular pH measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ. Biochemistry 18, 22102218.
Wendt, MK and Schiemann, WP (2009) Therapeutic targeting of the focal adhesion complex prevents oncogenic TGF-β signaling and metastasis. Breast Cancer Res 11, R68.
Wiemer, AJ, Wernimont, SA, Cung, T.-D, Bennin, DA, Beggs, HE and Huttenlocher, A (2013) The focal adhesion kinase inhibitor PF-562,271 impairs primary CD4+ cell activation. Biochem Pharmacol 86, 770781.
Xu, B, Zhou, C, Meredith, M and Baltz, JM (2017) Acute cell volume regulation by Janus kinase 2-mediated sodium/hydrogen exchange activation develops at the late one-cell stage in mouse preimplantation embryos. Biol Reprod 96, 542550.
Yoon, H, Choi, Y.-L, Song, J.-Y, Do, I, Kang, SY, Ko, Y.-H, Song, S and Kim, B.-G (2014) Targeted inhibition of FAK, PYK2 and BCL-XL synergistically enhances apoptosis in ovarian clear cell carcinoma cell lines. PLoS One 9, e88587.
Zhou, C and Baltz, JM (2013) JAK2 mediates the acute response to decreased cell volume in mouse preimplantation embryos by activating NHE1. J Cell Physiol 228, 428438.
Zhou, C, FitzHarris, G, Alper, SL and Baltz, JM (2013) Na+/H+ exchange is inactivated during mouse oocyte meiosis, facilitating glycine accumulation that maintains embryo cell volume. J Cell Physiol 228, 20422053.


Related content

Powered by UNSILO

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

  • Jane C. Fenelon (a1) (a2), Baozeng Xu (a1) (a2) (a3) and Jay M. Baltz (a1) (a2)


Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.