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

海豚细胞培养方法的建立及新型水通道蛋白的功能研究

• 批准号: 23580265
• 负责人: SUZUKI Miwa
• 金额: $ 3.49万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2013
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23580265/
• 关键词: 鯨類; 浸透圧調節; 細胞培養; アクアポリン; 海水適応; イルカ; 浸透圧耐性; ノックダウン; バンドウイルカ; 浸透圧

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.

我们之前的数据表明，瓶鼻海豚体内存在水通道蛋白(AQP)-2的选择性剪接。由于aqp2基因对高渗有反应元件，我们推测交替调味的AQP2(AQP2‘)具有一定的细胞渗透调节功能。对这一假说进行了实验验证，得到了有趣的结果：(1)建立了海豚几个器官的原代细胞培养技术。(2)AQP2仅在肾脏中表达，而AQP2在所有受试器官中均有表达。(3)基因组分析表明，AQP2‘序列在鲸目动物中高度保守，在其他哺乳动物中不存在。(4)AQP2和AQP2‘的表达随着培养液中Na+浓度的升高而增加。(5)小干扰RNA对AQP2和AQP2‘的敲除导致海豚细胞严重萎缩，细胞存活率降低。总而言之，这些数据表明，AQP2可能有助于细胞对渗透压的耐受。

项目成果

Molecular characterization and validation of commercially available methods for haptoglobin measurement in bottlenose dolphin

宽吻海豚触珠蛋白测量市售方法的分子表征和验证

• 发表时间: 2013
• 期刊: Results in Immunology
• 作者: Segawa T;Amatsuji H;Suzuki K;Suzuki M;Yanagisawa M;Itou T;Sakai T;Nakanishi T.
• 通讯作者: Nakanishi T.

"鯨類は水を飲まないのか?—浸透圧調節—"『ケトスの知恵』,村山司・森阪匡通編著

《鲸鱼不喝水吗？-渗透压调节-》《酮的智慧》，村山司、森坂正通编辑

• 发表时间: 2012
• 作者: S. Fujinaga;I. Yoshinaga;M. Tanaka;杉田治男（編）;大越健嗣;鈴木美和;吉永郁生;鈴木美和
• 通讯作者: 鈴木美和

Characterization of the circulating serum amyloid A in bottlenose dolphins.

宽吻海豚循环血清淀粉样蛋白 A 的表征。

• DOI: 10.1016/j.vetimm.2012.12.006
• 发表时间: 2013
• 期刊: Vet Immunol Immunopathol.
• 作者: Segawa T;Otsuka T;Itou T;Suzuki M;Karatani N;Sakai T.
• 通讯作者: Sakai T.

水族館と海の生き物たち

水族馆和海洋生物

• 发表时间: 2014
• 作者: 牧口祐也;北山武憲;根本武;市村政樹;小島隆人;牧口祐也
• 通讯作者: 牧口祐也

Activation of systemic, but not local, renin-angiotensin system is associated with upregulation of TNF-α during prolonged fasting in northern elephant seal pups.

北象海豹幼崽长期禁食期间，全身而非局部肾素-血管紧张素系统的激活与 TNF-α 的上调有关。

• DOI: 10.1242/jeb.085225
• 发表时间: 2013
• 期刊: The Journal of experimental biology
• 作者: Suzuki,Miwa;Vázquez-Medina,JoséPablo;Viscarra,JoseA;Soñanez-Organis,JoséG;Crocker,DanielE;Ortiz,RudyM
• 通讯作者: Ortiz,RudyM