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a Department of Animal Health and Biomedical Sciences, University of Wisconsin, Madison, Wisconsin 53706
b Loeb Research Institute and Departments of Obstetrics and Gynecology and Cellular and Molecular Medicine, University of Ottawa, Ontario, Canada K1Y 4E9
The ability to regulate intracellular pH (pHi) is essential for normal cell development and differentiation. This study was an investigation of the regulatory system used by the hamster oocyte and preimplantation embryo to regulate pHi in the alkaline range. Recovery from alkalosis by late 1-cell and 2-cell embryos was rapid, and physiological pHi levels could be restored within 10 min. Recovery from an induced alkaline load was dependent on the chloride concentration in the external medium and sensitive to a stilbene derivative 4,4'-diisothiocyanatostilbene-2,2'-di-sulfonic acid that inhibits bicarbonate and chloride exchange. Therefore the recovery from alkalosis by hamster embryos appears to be via activity of the HCO3-/Cl- exchanger that was activated above a pHi set point of 7.24. In contrast, hamster oocytes and early 1-cell embryos (collected 3–4 h post-egg activation) could not recover from an intracellular alkalosis, and pHi remained elevated. Therefore, the hamster oocyte and the early 1-cell embryo still undergoing pronuclear formation lack an active HCO3-/Cl- exchanger for the restoration of pHi. Inability to restore pHi from an alkali challenge resulted in a reduced ability of embryos to develop to the morula/blastocyst stages in culture, indicating that HCO3-/Cl- exchange is involved in physiological regulation of pHi.
2 Correspondence and current address: Michelle Lane, 799 East Hampden Ave., Suite 300, Englewood, CO 80110. FAX: 303 788 4438; mlane{at}colocrm.com
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