Osmometric behavior of mouse oocytes in the presence of different intracellular sugars.
In order to successfully cryopreserve oocytes using low concentrations of intracellular sugars, it is important to characterize their osmotic response in the presence of these intracellular sugars. In the present study, murine (B6D2F1) oocytes were microinjected with 0.8M glucose, trehalose or stachyose solutions to achieve an intracellular concentration equivalent to 0.1M, and then exposed to hypertonic solutions of increasing strength by supplementing an isotonic solution with 0.2, 0.3, 0.5, and 0.7 M of trehalose. Analysis of volumetric response of microinjected oocytes showed that the oocytes behaved as ideal osmometers in the presence of intracellular sugars and satisfied the Boyle van't Hoff relationship. Extrapolation of the osmotically inactive fraction (V macro b) from the Boyle van't Hoff relationship yielded values of 0.188+/-0.028, 0.212+/-0.042, 0.197+/-0.044, and 0.211+/-0.042 for control, glucose, trehalose and stachyose-injected oocytes, respectively. The present data revealing osmometric behavior of mouse oocytes in the presence of different intracellular sugars are important for the optimization of cryopreservation protocols using sugars.
Bhowmick
Pragati
P
Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School and Shriners Hospital for Children, Boston, MA 02114, USA.
Eroglu
Ali
A
Wright
Diane L
DL
Toner
Mehmet
M
Toth
Thomas L
TL
D000818:Animals; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000008:administration & dosage; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000008:administration & dosage / Q000378:metabolism; D005260:Female; D005947:Glucose / Q000008:administration & dosage / Q000378:metabolism; D066298:In Vitro Techniques; D007424:Intracellular Fluid / Q000378:metabolism; D051379:Mice; D008845:Microinjections; D009844:Oligosaccharides / Q000008:administration & dosage / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism*; D009994:Osmolar Concentration; D009995:Osmosis; D014199:Trehalose / Q000008:administration & dosage / Q000378:metabolism
D002241:Carbohydrates; D003451:Cryoprotective Agents; D009844:Oligosaccharides; C005695:stachyose; D014199:Trehalose; D005947:Glucose