Intracellular concentrations of potassium and sodium in two-cell mouse embryos in G1/S phase after exposure to vitrification solutions containing vitrificant agents (VFAs) ethane-1,2-diol (ethylene glycol, EG), propane-1,2-diol (propylene glycol, PG), or dimethyl sulfoxide (DMSO), and sucrose (S) after incubation in Dulbecco's solution were measured by electron probe microanalysis (EPMA) as described earlier (CryoLetters, 2006, 27:87-98). The 4-step protocol was as followed: 10%-VFA for 10 min ⇒ 30%-VFA + 0.7 M S for 1.5 min ==> 0.5 M S for 10 min ==> 10 min pure Dulbecco's. The cytoplasmic concentration of potassium and sodium in immediately flashed out from the oviduct embryos was in range of 120 ± 2 mM, with good concordance with the previous data (CryoLetters, 2006, 27:87-98). Exposure in Dulbecco's for 30 min did not alter elemental composition, neither did exposure in PG or DMSO for 1.5 min. In contrast, exposure for 1.5 min in EG dropped the level of potassium to 96 ± 2 mM while elevating level of cytoplasmic sodium to 136 ± 3 mM. Further exposure to 30%-EG for 3 min led to a two-fold decrease of both elements (60 ± 3 mM and 66 ± 2 mM for K and Na respectively).
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EARLY MOUSE EMBRYO;
ELECTRON PROBE MICROANALYSIS (EPMA);
INTRACELLULAR POTASSIUM AND SODIUM;
Document Type: Research Article
November 1, 2007
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CryoLetters is a bimonthly international journal for low temperature sciences, including cryobiology, cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and studies involving ecology of cold environments, and cold adaptation
The journal publishes original research reports, authoritative reviews, technical developments and commissioned book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical processes, or those involving low temperature techniques in the investigation of physical, chemical, biological and ecological problems.