Phosphatidylinositol 4-kinase β is required for the ciliogenesis of zebrafish otic vesicle
Phosphatidylinositol 4-kinase β is required for the ciliogenesis of zebrafish otic vesicle
复制标题
斑马鱼耳囊纤毛发生需要磷脂酰肌醇 4 激酶-β
DOI：
10.1016/j.jgg.2020.07.007
复制
发表时间：
2020-10-20
影响因子：
5.9
通讯作者：
Zhang, Jingjing
中科院分区：
文献类型：
--
作者：
Feng, Yufei;Yu, Ping;Zhang, Jingjing
初级纤毛是一种重要的基于微管的细胞器，几乎从所有脊椎动物细胞中伸出。纤毛的运动对于各种发育和生理过程是必需的。磷酸肌醇 (PI) 及其代谢物 PtdIns(4,5)P2 已被发现有助于纤毛组装和分解。作为 PI 通路和信号传导的重要激酶，磷脂酰肌醇 4-激酶 β (PI4KB) 是研究最广泛的磷脂酰肌醇 4-激酶亚型之一。然而，它在器官发育中的潜在作用仍有待表征。为了研究 Pi4kb 的发育作用，特别是其在斑马鱼纤毛发生中的功能，我们使用成簇规则间隔短回文重复序列 (CRISPR)/CRISPR 相关蛋白 9 技术生成了 pi4kb 缺失突变体。纯合的 pi4kb 突变体表现出内耳、神经丘和原肾管中初级纤毛的缺失，并伴有眼睛和其他器官的严重水肿。此外，较小的耳泡、畸形的半规管以及对声音刺激不敏感是pi4kb突变体的特征。在蛋白质水平上，体内和体外分析均表明，由于 Pi4kb 的丢失，Pi4p 的合成大大减少。此外，在 pi4kb 突变体中，神经元钙传感器 1 的 Pi4kb 结合伴侣的表达以及 Akt 下游效应器 4-磷酸磷脂酰肌醇的磷酸化均受到显着抑制。总而言之，我们的工作通过确定毛细胞纤毛发生，揭示了 Pi4kb 在斑马鱼内耳发育和听力功能形成中的新作用。版权所有(C)2020，中国科学院遗传与发育生物学研究所，中国遗传学会。由爱思唯尔有限公司和科学出版社出版。版权所有。
The primary cilium, an important microtubule-based organelle, protrudes from nearly all the vertebrate cells. The motility of cilia is necessary for various developmental and physiological processes. Phosphoinositides (PIs) and its metabolite, PtdIns(4,5)P2, have been revealed to contribute to cilia assembly and disassembly. As an important kinase of the PI pathway and signaling, phosphatidylinositol 4-kinase beta (PI4KB) is the one of the most extensively studied phosphatidylinositol 4-kinase isoform. However, its potential roles in organ development remain to be characterized. To investigate the developmental role of Pi4kb, especially its function on zebrafish ciliogenesis, we generated pi4kb deletion mutants using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 technique. The homozygous pi4kb mutants exhibit an absence of primary cilia in the inner ear, neuromasts, and pronephric ducts accompanied by severe edema in the eyes and other organs. Moreover, smaller otic vesicle, malformed semicircular canals, and the insensitivity on sound stimulation were characteristics of pi4kb mutants. At the protein level, both in vivo and in vitro analyses revealed that synthesis of Pi4p was greatly reduced owing to the loss of Pi4kb. In addition, the expression of the Pi4kb-binding partner of neuronal calcium sensor-1, as well as the phosphorylation of phosphatidylinositol-4-phosphate downstream effecter of Akt, was significantly inhibited in pi4kb mutants. Taken together, our work uncovers a novel role of Pi4kb in zebrafish inner ear development and the functional formation of hearing ability by determining hair cell ciliogenesis. Copyright (C) 2020, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, and Genetics Society of China. Published by Elsevier Limited and Science Press. All rights reserved.