Cryoprotectant permeability of aquaporin-3 expressed in Xenopus oocytes
Cryoprotectant permeability of aquaporin-3 expressed in Xenopus oocytes
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DOI：
10.1016/j.cryobiol.2006.06.008
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发表时间：
2006-10-01
期刊：
影响因子：
2.7
通讯作者：
Edashige, Keisuke
中科院分区：
文献类型：
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作者：
Yamaji, Yohei;Valdez, Delgado M., Jr.;Edashige, Keisuke
研究表明，水通道蛋白-3在小鼠胚胎中表达。这种类型的水通道蛋白不仅运输水，还运输中性溶质，包括渗透细胞的冷冻保护剂。因此，该通道的表达可能对冷冻胚胎的存活有重要影响。然而，除甘油外，水通道蛋白-3对低温保护剂的渗透系数尚未测定。此外，浓度梯度下的渗透系数对于制定和改进低温保存方案具有重要意义。在这项研究中，我们用非洲爪蟾卵细胞检测了水通道蛋白-3对各种冷冻保护剂的渗透性。在含有10%甘油、8%乙二醇、10%丙二醇的改性Barth's溶液中，通过注射水通道蛋白-3 rna的卵母细胞的体积变化来测量水通道蛋白-3对冷冻保护剂的渗透性。水通道蛋白-3对乙二醇和丙二醇的渗透系数分别为33.50和31.45 × 10(-3) cm/min，与对甘油的渗透系数（36.13 × 10(-3) cm/min）相当。这些数值远高于注射水的对照卵母细胞（0.04 ~ 0.11 × 10(-3) cm/min）。另一方面，乙酰胺和DMSO的系数不能很好地确定，因为两参数模型的体积数据不能很好地拟合，可能是由于膜损伤。为了避免这种情况，在低浓度梯度下，通过将卵母细胞悬浮在含有低浓度乙酰胺或DMSO溶解在水中（0.20 Osm/kg）的水溶液中来测量这些冷冻保护剂的渗透性。乙酰胺的系数（24.60 × 10(-3) cm/min）与甘油、乙二醇和丙二醇的系数一样高，显著高于对照组的系数（6.50 × 10(-3) cm/min）。DMSO值（6.33 × 10(-3) cm/min）相对较低，但高于对照组（0.79 × 10(-3) cm/min）。这是首次报道的通过水通道蛋白-3转运DMSO的观察结果。(c) 2006爱思唯尔公司版权所有。
It has been shown that aquaporin-3, a water channel, is expressed in mouse embryos. This type of aquaporin transports not only water but also neutral solutes, including cell-permeating cryoprotectants. Therefore, the expression of this channel may have significant influence on the survival of cryopreserved embryos. However, permeability coefficients of aquaporin-3 to cryoprotectants have not been determined except for glycerol. In addition, permeability coefficients under concentration gradients are important for developing and improving cryopreservation protocols. In this study, we examined the permeability of aquaporin-3 to various cryoprotectants using Xenopus oocytes. The permeability of aquaporin-3 to cryoprotectants was measured by the volume change of aquaporin-3 cRNA-injected oocytes in modified Barth's solution containing either 10% glycerol, 8% ethylene glycol, 10% propylene glycol,(.) 1.5 M acetamide, or 9.5% DMSO (1.51-1.83 Osm/kg) at 25 degrees C. Permeability coefficients of aquaporin-3 for ethylene glycol and propylene glycol were 33.50 and 31.45 x 10(-3) cm/min, respectively, which were as high as the value for glycerol (36.13 x 10(-3) cm/min). These values were much higher than those for water-injected control oocytes (0.04-0.11 X 10(-3) cm/min). On the other hand, the coefficients for acetamide and DMSO were not well determined because the volume data were poorly fitted by the two parameter model, possibly because of membrane damage. To avoid this, the permeability for these cryoprotectants was measured under a low concentration gradient by suspending oocytes in aqueous solutions containing low concentrations of acetamide or DMSO dissolved in water (0.20 Osm/kg). The coefficient for acetamide (24.60 x 10(-3) cm/min) was as high as the coefficients for glycerol, ethylene glycol, and propylene glycol, and was significantly higher than the value for control (6.50 x 10(-3) cm/min). The value for DMSO (6.33 x 10(-3) cm/min) was relatively low, although higher than the value for control (0.79 x 10(-3) cm/min). This is the first reported observation of DMSO transport by aquaporin-3. (c) 2006 Elsevier Inc. All rights reserved.