Developmental expression of aquaporin-3 in zebrafish embryos (Danio rerio)
Developmental expression of aquaporin-3 in zebrafish embryos (Danio rerio)
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DOI：
10.1016/j.cca.2004.08.004
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发表时间：
2004-07-01
影响因子：
3.9
通讯作者：
Hagedorn, M
中科院分区：
文献类型：
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作者：
Lance, SL;Peterson, AS;Hagedorn, M
由于冷冻保护剂对卵黄的渗透性低，鱼类胚胎从未成功地冷冻保存过。最近，我们使用水通道蛋白3融合绿色荧光蛋白（AQP 3 GFP）修饰斑马鱼胚胎，并证明了孔生理功能。这增加了膜的水和冷冻保护剂渗透性。我们继续对AQP 3修饰的胚胎进行研究，在这里我们报告了AQP 3的发育表达，各种培养基对其生存和发育的成功，以及它们的繁殖成功。AQP 3GFP的表达开始于mRNA AQP 3GFP注射到1- 4-细胞胚胎的卵黄后30 m内。这种表达在24小时内分布在整个胚盘和卵黄合胞体层的膜中。96 h后下降。我们发现AQP 3GFP或野生型成人胚胎的存活或正常发育没有差异。此外，斑马鱼胚胎在AQP 3GFP修饰后不需要特殊的培养基即可存活。事实上，它们在胚胎培养基中存活得最好（约。40 mOsm）。完全在胚胎培养基中培养的胚胎比暴露于高渗透压蔗糖培养基的胚胎具有更高的存活率和更高的正常发育率（约10%）。330 mOsm）。这些胚胎能够在低渗溶液中保持其内部渗透压的机制尚不清楚。(C)2004年爱思唯尔公司All rights reserved.
Fish embryos have never been successfully cryopreserved because of the low permeability of cryoprotectants into the yolk. Recently, we used aquaporin-3 fused with a green fluorescent protein (AQP3GFP) to modify the zebrafish embryo, and demonstrated that the pores functioned physiologically. This increased the water and cryoprotectant permeability of the membranes. We have continued our work on AQP3-modified embryos and here we report their developmental expression of AQP3, the success of various culture media on their survival and development, and their reproductive success. The AQP3GFP expression begins within 30 m after the mRNA AQP3GFP injection into the yolk of the 1- to 4-cell embryo. This expression is distributed in the membranes throughout the blastoderm and the yolk syncytial layer within 24 h. It diminishes after 96 h. We found no difference in the survival or normal development of embryos from AQP3GFP or wild-type adults. Additionally, zebrafish embryos did not require special culture medium to survive after AQP3GFP modification. In fact, they survived best in embryo medium (ca. 40 mOsm). Embryos reared entirely in embryo medium had a higher percent survival and a higher percent normal development than those exposed to a high osmolatity sucrose culture medium (ca. 330 mOsm). The mechanism whereby these embryos can maintain their internal osmolality in a hypoosmotic solution with water channels in their membranes is unknown. (C) 2004 Elsevier Inc. All rights reserved.