Compensatory response of blastomeres in two-cell mouse embryo to hypoosmotic stress was studied employing the direct measurement of cellular volume with Laser Scanning Microtomography followed by 3-D reconstruction (QLSM). It was shown that embryos exposed to hypotonicity first swelled and then returned to the initial size. Swelling phase was defined by a water permeability coefficient (Lp) of 0.4 micron/(min.atm). The next compensatory phase of regulatory volume decrease (RVD) in embryonic cells was not dependent on Na+/K+-ATPase inhibition or low pH (6.4), but RVD was abolished by Cytochalasine B (Cyto B) treatment.
two-cell mouse embryo, regulatory volume decrease, Na+/K+-ATPase, Cytochalasine B, laser scanning microscopy, 3D-reconstruction
1. Praissman M., Miller I.F., Berkowitz J.M. Ion-mediated water flow. I. Electroosmosis. J Membrane Biol., 1973, vol. 11, pp. 139-151.
2. Praissman M., Miller I.F., Gregor H.P., Berkowitz J.M. Ion-mediated water flow. II. Anomalous osmosis. J Membrane Biol., 1973, vol. 11, pp. 153-167.
3. Loeb J. On the cause of the influence of ions on the rate of diffusion of water through collodion membrane. J Gen Physiol., 1920, vol. II, pp. 387-408.
4. Grim E., Sollner K. The contribution of normal and anomalous osmosis to the osmotic effects arising across charged membranes with solution of electrolytes. J Gen Physiol., 1957, vol. 40, pp. 887-889.
5. Buckhold B., Adams R.B., Gregg E.C. Osmotic adaptation of mouse lymphoblasts. Biochim Biophys Acta, 1965, vol. 102, pp. 600-608.
6. Rosenberg H.M., Buckhold Shank B., Gregg E.C. Volume changes of mammalian cells subjected to hypotonic solution in vitro: evidence for the requirement of sodium pump for the shrinking phase. J Cell Physiol., 1972, vol. 80, pp. 23-32.
7. Buckhold Shank B., Rosenberg H.M., Horowitz C. Ionic basis of volume regulation in mammalian cells following osmotic shock. J Cell Physiol., 1973, vol. 82, pp. 257-266.
8. Pogorelov A.G., Allachverdov B.L., Burovina I.V., Mazay G.G., Pogorelova V.N. Study of potassium deficiency in cardiac muscle: quantitative X-ray microanalysis and cryotechniques. J Microsc., 1991, vol. 162, pp. 255-269.
9. Pogorelov A.G., Pogorelova V.N., Repin N.V., Mizin I.A. Quantitative electron probe microanalysis with a wavelength dispersive spectrometer. Scanning Microsc., 1994, Suppl. 8, pp. 101-108.
10. Pogorelov A.G., Katkov I.I., Pogorelova V.N. Influence of exposure to vitrification solutions on 2-cell mouse embryos: I. Intracellular potassium and sodium content. CryoLetters, 2007, vol. 28, pp. 403-408
11. Pogorelov A.G., Pogorelova V.N. Quantitative tomography of early mouse embryos: Laser Scanning Microscopy and 3-D reconstruction. J Microsc., 2008, vol. 232, pp. 36-43.
12. Pogorelova M.A., Golichenkov V.A., Pogorelov A.G. Differential Axial Contrast of Optical Sections: Laser Microtomographyand Quantitative 3D Reconstruction. Optics and Spectrosc., 2014, vol. 116, pp. 488-493.
13. Fong B., Watson P.H., Watson A.J. Mouse preimplantation embryo responses to culture medium osmolarity include increased expression of CCM2 and p38 MAPK activation. BMC Dev Biol., 2007, vol. 7, pp. 1-16.
14. Duque P., Hidalgo C.O., Gomez E., Pintado B., Facal N., Diez C. Macromolecular source as dependent on osmotic pressure and water source: effects on bovine in vitro embryo development and quality. Reprod Nutr Dev., 2003, vol. 43, pp. 487-496.
15. Gao D.Y., McGrath J.J., Tao J., Benson C.T., Critser E.S., Critser J.K. Membrane transport properties of mammalian oocytes: a micropipette perfusion technique. J Reprod Fertil., 1994, vol. 102, pp. 385-392.
16. Gao D.Y., Benson C.T., Liu C., McGrath J.J., Critser E.S., Critser J.K. Development of a novel microperfusion chamber for determination of cell membrane transport properties. Biophys J., 1996, vol. 71, pp. 443-450.
17. Agca Y., Liu J., McGrath J.J., Peter A.T., Critser E.S., Critser J.K. Membrane permeability characteristics of metaphase II mouse oocytes at various temperatures in the presence of Me2SO. Cryobiology, 1998, vol. 36, pp. 287-300.
18. Watson A.J., Kidder G.M. Immunofluorescence assessment of the timing of appearance and cellular distribution of Na/K-ATPase during mouse embryogenesis. Dev Biol., 1998, vol. 126, pp. 80-90.
19. Van Winkle L.J., Campione A.L. Ouabain-sensitive Rb+ uptake in mouse eggs and preimplantation conceptuses. Dev Biol., 1991, vol. 146, pp. 158-166.
20. Slack C., Warner A.E., Warren R.L. The distribution of sodium and potassium in amphibian embryos during early development. J Physiol., 1973, vol. 232, pp. 297-312.
21. Verkman A.S., Mitra A.K. Structure and function of aquaporin water channels. Am J Physiol Renal Physio., 2000, vol. 278, pp. F13-F28.
22. Agre P. The aquaporin water channels. Proc Am Thorac Soc., 2005, vol. 3, pp. 5-13.
23. Cahalin K.L., Hall J.E. pH sensitivity of MIP-induced water permeability may play a role in regulating the intrinsic fluid circulation of the lens. Biophys J., 1999, vol. 76, pp. 183-189.
24. Yasui M., Kwon T.H., Knepper M.A., Nielsen S., Agre P. Aquaporin 6: an intracellular vesicle water channel protein in renal epithelia. Proc Natl Acad Sci USA, 1999, vol. 96, pp. 5808-5813.
25. Baltz J.M., Biggers J.D., Lechene C. A novel H+ permeability dominating intracellular pH in the early mouse embryo. Development, 1993, vol. 118, pp. 1353-1361.
26. Hoffmann E.K., Simonsen L.O. Membrane mechanisms in volume and pH regulation in vertebrate cells. Physiol Rev., 1989, vol. 69, pp. 315-388.
27. Lang F., Bush G.L., Ritter M., Volkl H., Waldegger S., Gulbins E., Haussinger D. Functional significance of cell volume regulatory mechanisms. Physiol Rev., 1998, vol. 78, pp. 248-306.
28. Perry M.M., Snow M.H. The blebbing response of 2-4 cell stage mouse embryos to cytochalasin B. Dev Biol., 1975, vol. 45, pp. 372-377.
29. Pogorelova M.A., Golichenkov V.A., Tarasov A.V., Pogorelova V.N., Panait A.I., Pogorelov A.G. Cyto B Dependent and Ouabain Insensitive Regulatory Volume Decrease in Bicellular Mouse Embryo. Russian J Develop. Biol., 2012, vol. 43, pp. 83-90.
30. Samoilov O.Y. Structure of aqueous electrolyte solutions and hydration of ions. Consultants Bureau, Enterpr. Inc., New York, 1965, 185 p.