Harvey B, Kelley R N, Ashwood-Smith M J
Cryobiology. 1983 Aug;20(4):432-9. doi: 10.1016/0011-2240(83)90033-0.
Intact developing embryos of the zebra fish Brachydanio rerio were exposed to [14C]DMSO and [3H]glycerol (1 M in Fish Ringer) to assess the degree of permeation of these cryoprotectants. Glycerol enters the embryo more easily, although reaching only about 8% of the expected equilibrium level after 2 hr at room temperature; DMSO reaches only about 2.5% of this level. In an attempt to identify the barrier to permeation, dechorionated embryos were similarly exposed to isotopic DMSO. Permeation increased severalfold, indicating that the chorion retards the free exchange of solute. Embryos are unaffected by exposure to 1 M DMSO in Fish Ringer at 23 degrees C for up to 1 hr. The number of embryos hatching after 1-hr exposure to DMSO at varying concentrations was significantly reduced at 1.5 and 2 M. Embryos exposed to 1 M glycerol for 1 hr at 23 degrees C showed disruption of periblast cells and separation of the blastoderm; it was impossible to remove glycerol either by abrupt or very slow dilution.
将斑马鱼（Brachydanio rerio）完整的发育胚胎暴露于[14C]二甲基亚砜（DMSO）和[3H]甘油（在鱼类任氏液中浓度为1 M）中，以评估这些冷冻保护剂的渗透程度。甘油更容易进入胚胎，尽管在室温下放置2小时后仅达到预期平衡水平的约8%；DMSO仅达到该水平的约2.5%。为了确定渗透的障碍，将去绒毛膜的胚胎同样暴露于同位素DMSO中。渗透率增加了几倍，表明绒毛膜阻碍了溶质的自由交换。胚胎在23摄氏度下暴露于鱼类任氏液中的1 M DMSO中长达1小时不受影响。在1.5 M和2 M浓度下，暴露于不同浓度DMSO 1小时后孵化的胚胎数量显著减少。在23摄氏度下将胚胎暴露于1 M甘油中1小时，显示出胚周细胞的破坏和胚盘的分离；无论是通过突然稀释还是非常缓慢的稀释都无法去除甘油。
