Functional challenge affects aquaporin rnRNA abundance in mouse Blastocysts
Functional challenge affects aquaporin rnRNA abundance in mouse Blastocysts
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DOI：
10.1002/mrd.20306
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发表时间：
2005-08-01
影响因子：
2.5
通讯作者：
Thomsen, PD
中科院分区：
文献类型：
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作者：
Offenberg, H;Thomsen, PD
水通道蛋白（aquaporins，AQP）是一类促进水分子跨膜扩散的通道蛋白。在小鼠胚泡中检测到AQP家族的三个成员：AQP 3和8位于滋养层细胞的基底外侧区，AQP 9主要位于滋养层细胞的顶端区。据信这些参与促进液体积聚到胚泡腔中。我们研究了小鼠胚胎调节AQP基因表达的能力，以应对不同的治疗方法，预计会影响水通过滋养层细胞，使用实时PCR。在第一个实验中，允许8细胞胚胎在300至400 mOsm的培养基中发育成囊胚。囊胚形成不受甘油高渗培养基的影响，而在蔗糖基350和400 mOsm培养基中囊胚形成显著减少。当胚胎在甘油基高渗培养基中培养时，AQP 8 mRNA水平降低。AQP 3、7、9和11的mRNA水平不受高渗介质的显著影响。在第二个实验中，将胚泡穿刺（0小时）并使其再扩张。与对照胚胎相比，AQP 3、7和9的表达在2小时后上调，仅AQP 9的表达持续上调，并且这种上调持续到穿刺后6和10小时。在第三个实验中，我们比较了水通道蛋白在体外培养和体内发育的囊胚中的表达。我们发现，在体外培养导致较低水平的AQP 8，9和11相比，在体内发展。这些实验表明，小鼠胚胎能够调节AQP mRNA丰度，以响应环境的改变。
The aquaporins (AQPs) are a family of channel proteins that facilitate diffusion of water across cell membranes. Three members of the AQP family have been detected in the mouse blastocyst: AQP 3 and 8 are located in the basolateral domain and AQP 9 predominantly in the apical domain of the trophoblast cells. These are believed to be involved in facilitating the accumulation of fluid into the blastocyst cavity. We have investigated the ability of mouse embryos to regulate AQP gene expression in response to different treatments expected to affect the passage of water across the trophoblast cells using real-time PCR. In the first experiment 8-cell embryos were allowed to develop to blastocysts in media from 300 to 400 mOsm. Blastocyst formation was unaffected by media made hyperosmolar by glycerol, whereas blastocyst formation was significantly reduced in sucrose-based 350 and 400 mOsm media. AQP 8 mRNA levels were reduced when embryos were cultured in glycerol-based hyperosmolar media. The mRNA levels of AQP 3, 7, 9, and 11 were not significantly affected by hyperosmolar media. In the second experiment blastocysts were punctured (0 hr) and allowed to re-expand. AQP mRNA levels were examined after 2, 6, and 10 hr. Compared to control embryos, the expression of AQP 3, 7, and 9 were upregulated after 2 hr. Upregulation was sustained only for AQP 9 and this was sustained up to 6 and 10 hr after puncture. In the third experiment we compared expression of AQPs between in vitro cultured and in vivo developed blastocysts. We found that in vitro culture resulted in lower levels of AQP 8, 9, and 11 compared to in vivo development. These experiments show that mouse embryos are capable of regulating AQP mRNA abundances in response to environmental alterations.