Exogenous expression of rat aquaporin-3 enhances permeability to water and cryoprotectants of immature oocytes in the zebrafish (Danio rerio)
Exogenous expression of rat aquaporin-3 enhances permeability to water and cryoprotectants of immature oocytes in the zebrafish (Danio rerio)
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DOI：
10.1262/jrd.18164
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发表时间：
2007-06-01
影响因子：
1.8
通讯作者：
Edashige, Keisuke
中科院分区：
文献类型：
--
作者：
Seki, Shinsuke;Kouya, Toshimitsu;Edashige, Keisuke
由于斑马鱼的卵母细胞/胚胎比哺乳动物的卵母细胞/胚胎要大得多，因此需要加速水和冷冻保护剂通过质膜的运动。水通道蛋白-3是一种水/溶质通道，不仅可以运输水，还可以运输各种冷冻保护剂。在这项研究中，我们试图增加未成熟的斑马鱼卵母细胞在III期对水和冷冻保护剂的渗透性；通过外源性大鼠水通道蛋白3的表达。将未成熟的斑马鱼卵母细胞注射大鼠水通道蛋白-3 cRNA，培养5-12小时，根据卵母细胞在25℃高渗蔗糖溶液和各种冷冻保护剂溶液中体积的变化来测定其对水和冷冻保护剂的渗透性。注射水通道蛋白-3 cRNA的卵母细胞对水的渗透性比注射完整卵母细胞和水的卵母细胞高3倍。注射水通道蛋白- 3crna的卵母细胞对乙二醇、甘油、丙二醇和DMSO的渗透性也比完整卵母细胞高2-4倍。因此，外源表达水通道蛋白-3可以增强未成熟斑马鱼卵母细胞对水和冷冻保护剂的渗透性。硬骨卵母细胞的低温保存可以通过进一步增加通透性来实现。
Movement of water and cryoprotectants through the plasma membrane needs to be accelerated for successful cryopreservation of zebrafish oocytes/embryos, which are much larger than their mammalian counterparts. Aquaporin-3 is a water/solute channel that can transport not only water but also various cryoprotectants. In this study, we attempted to increase the permeability of immature zebrafish oocytes at stage III to water and cryoprotectants; by exogenous expression of rat aquaporin-3. Immature zebrafish oocytes were injected with rat aquaporin-3 cRNA and cultured for 5-12 h. Permeability to water and cryoprotectants was then determined based on changes in the volumes of the oocytes in a hypertonic sucrose solution and various cryoprotectant solutions at 25 C. The permeability to water of the aquaporin-3 cRNA-injected oocytes was three times higher than that of intact and water-injected oocytes. The permeability of the aquaporin-3 cRNA-injected oocytes to ethylene glycol, glycerol, propylene glycol, and DMSO was also 2-4 times higher than that of intact oocytes. Thus, the permeability of immature zebrafish oocytes to water and cryoprotectants was enhanced by exogenous expression of aquaporin-3. Cryopreservation of teleost oocytes may be realized through a further increase in permeability.