Ion-transporting ability and functional differentiation of chloride cells in the yolk-sac membrane of"yolk balls"

“卵黄球”卵黄囊膜氯细胞的离子传输能力及功能分化

• 批准号: 14360106
• 负责人: KANEKO Toyoji
• 金额: $ 7.36万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14360106/
• 关键词: mitochondrion-rich cell; yolk ball; Mozambique tilapia; Na^+; K^+-ATPase; K^+; 2Cl^- cotransporter; CFTR; yolk-sac membrane; embryo; アクアポリン; 水銀; 浸透圧変化感知機構; 免疫組織化学; Na^+; K^+-ATPase; 銀染色

We have recently established a unique in vitro experimental model for mitochondrion-rich cell(MRC) research, a 'yolk-ball' incubation system, in which the yolk sac is separated from the embryonic body of Mozambique tilapia embryos and subjected to in vitro incubation. To evaluate the ion-transporting property of the yolk balls, we examined Cl^- content and turnover in yolk balls incubated in freshwater and seawater for 48 h, and distribution patterns of three ion transporters, Na^+/K^+-ATPase, Na^+/K^+/2Cl^- cotransporter (NKCC) and cystic fibrosis transmembrane conductance regulator (CFTR), in MRCs in the yolk-sac membrane. The Cl^- turnover rate measured by whole-body influx of ^<36>Cl^- was about 60 times higher in yolk balls in seawater than in freshwater, while there was no essential difference in Cl^- content between them. Na^+/K^+-ATPase-immunoreactive MRCs were larger in yolk balls from seawater than yolk balls from freshwater. Distribution patterns of iontransporting proteins allowed us to classify MRCs in freshwater yolk balls into three types : cells showing only basolateral Na^+/K^+-ATPase, cells showing basolateral Na^+/K^+-ATPase and apical NKCC, and cells showing basolateral Na+/K^+-ATPase and basolateral NKCC. The seawater yolk balls, on the other hand, were characterized by the appearance of MRCs possessing basolateral Na^+/K^+-ATPase, basolateral NKCC and apical CFTR. Those seawater-type MRCs were considered to secrete Cl^-through the CFTR-positive apical opening to cope with diffusional Cl^- influx. These findings indicate that the yolk balls preserve the Cl^- transporting property of intact embryos, ensuring the propriety of the yolk ball as an in vitro experimental model for the yolk-sac membrane that contains MRCs.

我们最近建立了一种独特的富线粒体细胞（MRC）研究的体外实验模型，一种“蛋黄球”培养系统，其中卵黄囊从莫桑比克罗非鱼胚胎的胚胎体中分离出来并进行体外培养。为了评价卵黄球的离子转运特性，我们检测了在淡水和海水中孵育48 h的卵黄球的Cl^-含量和周转率，以及三种离子转运体Na^+/K^+- atp酶、Na^+/K^+/2Cl^-共转运体（NKCC）和囊性纤维化跨膜传导调节剂（CFTR）在卵黄囊膜MRCs中的分布模式。海水中卵黄球的Cl^-周转率比淡水中卵黄球的Cl^-周转率高约60倍，而两者之间的Cl^-含量没有本质差异。海水卵黄球的Na^+/K^+- atp酶免疫反应性MRCs大于淡水卵黄球。离子转运蛋白的分布模式使我们能够将淡水卵黄球的MRCs分为三种类型：仅显示基底侧Na^+/K^+- atp酶的细胞，显示基底侧Na^+/K^+- atp酶和顶端NKCC的细胞，以及显示基底侧Na+/K^+- atp酶和基底侧NKCC的细胞。另一方面，海水蛋黄球的特征是出现具有基底侧Na^+/K^+- atp酶、基底侧NKCC和根尖CFTR的MRCs。海水型MRCs通过cftr阳性的根尖开口分泌Cl^-，以应对弥漫性Cl^-内流。这些发现表明，卵黄球保留了完整胚胎的Cl^-运输特性，确保了卵黄球作为含有MRCs的卵黄囊膜的体外实验模型的适宜性。

项目成果

Mechanism of acid adaptation of a fish living in a pH 3.5 lake

• DOI: 10.1152/ajpregu.00267.2002
• 发表时间: 2003-05-01
• 期刊: AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY
• 影响因子: 2.8
• 作者: Hirata, T;Kaneko, T;Hirose, S
• 通讯作者: Hirose, S

Ohta, N., Kaneko, T.他: "Damage to the gills, skin and other tissues by lysenin and the coelomic fluid of the earthworm Eisenia foetida in two teleosts"J.Exp.Zool.. 295A. 117-126 (2003)

Ohta, N., Kaneko, T. 等人：“两种硬骨鱼中的溶素和蚯蚓 Eisenia foetida 的体腔液对鳃、皮肤和其他组织的损害”J.Exp.Zool.. 117-126。 (2003)

Kaneko, T., Hasegawa, S., Sasai, S.: "Chloride cells in Japanese eel (Anguilla japonica) during their early life stages "Eel Biology" (K.Aida, K.Tsukamoto, and K.Yamauchi, Eds)"Springer. 497 (2003)

Kaneko, T.、Hasekawa, S.、Sasai, S.：“日本鳗鱼（Anguilla japonica）生命早期阶段的氯细胞“鳗鱼生物学”（K.Aida、K.Tsukamoto 和 K.Yamauchi，编辑）

Hatching enzymes in Japanese eel Anguilla japonica

日本鳗鱼的孵化酶

• 发表时间: 2004
• 期刊: Dev.Genes.Evol. 214
• 作者: Hiroi;J.;Maruyama;K.;Kawazu;K.;Kaneko;T.;Ohtani-Kaneko;R.;Yasumasu;S.
• 通讯作者: S.

Katoh F.: "Vacuolar-type proton pump in the basolateral plasma membrane energizes ion uptake in branchial mitochondria-rich cells of killifish"Journal of Experimental Biology. (印刷中). (2003)

Katoh F.：“基底外侧质膜中的液泡型质子泵为鳉鱼富含鳃线粒体的细胞中的离子摄取提供能量”实验生物学杂志（2003 年出版）。