The establishment of the method for cell culture and a study on the function of a new aquaporin in dolphins

我们之前的数据表明，水通道蛋白(AQP) -2在瓶鼻海豚中有选择性剪接。由于aqp2基因对高渗性有应答元件，我们推测aqp2 '具有一定的细胞渗透调节功能。对这一假设进行了实验验证，得到了有趣的结果：(1)建立了海豚多个器官原代细胞的培养技术。(2) AQP2仅在肾脏中表达，而AQP2在所有器官中均有表达。(3)基因组分析表明，AQP2序列在鲸类中高度保守，在其他哺乳动物中不保守。(4)培养基中Na+浓度增加，AQP2和AQP2′表达增加。(5)小干扰RNA敲低AQP2和AQP2'导致海豚细胞严重萎缩，细胞活力降低。总的来说，这些数据表明AQP2'可能有助于细胞的渗透耐受性。

Our previous data suggested alternative splicing of aquaporin (AQP) -2 in bottle nose dolphins. As aqp2 gene has response element to hypertonicity, we hypothesized the alternatively spiced-AQP2 (AQP2') has some functions for cellular osmoregulation. Experiments were carried out to test the hypothesis and interesting results were obtained: (1) Culture technique of primary cells from several organs of the dolphin was established. (2) AQP2 was expressed only in the kidney, whereas AQP2' expression was confirmed in all organs tested. (3) Genomic analyses showed AQP2' sequence is highly and broadly conserved in cetacean species, but not in other mammals. (4) Expression of AQP2 and AQP2' were increased in response to increase in Na+ concentration in medium. (5) Knockdown for AQP2 and AQP2' by small interfering RNA resulted in severe shrinkage and lowering cellular viability in dolphins' cells. Collectively, there data suggested AQP2' may contribute to cellular osmotic tolerance.