Exposure of Marmoset Oocytes to Electrical Stimulation
Twenty-four hours after collection, oocytes were subjected to electrical pulses generated by an Electro Cell Manipulator 200 (BTX Inc., San Diego, CA) applied in a chamber consisting of 2 parallel, stainless steel, 0.5-mm diameter electrodes attached to a glass slide, 0.5 mm apart (part no. 450; BTX Inc.). The chamber was filled with 0.3 M mannitol in embryo culture-quality water, supplemented with 100 ^M CaCl2 and 100 ^M MgCl2 (Sigma Chemical Co.). The oocytes were subjected to two series of six electrical pulses (DC), 30 min apart. Each pulse was 2 kV/cm and of 70-^sec duration. Control oocytes (n = 19) were not exposed to the mannitol solution or to electrical stimulation. proventil inhaler
Exposure of Marmoset Oocytes to Ethanol
Oocytes were incubated in 7% ethanol in PBS (Life Technologies, Paisley, Scotland) for 5 min at room temperature. Some of these oocytes (n = 28) had been incubated with sperm as part of another study. All oocytes in this group were exposed to ethanol 3 days after collection (4 days post-hCG), when it was confirmed that oocytes had not been fertilized, by absence of pronuclei and lack of cleavage for 48 h postinsemination.