Rab8, Rab11, and Rab35 coordinate lumen and cilia formation during zebrafish left-right organizer development.
Rab8, Rab11, and Rab35 coordinate lumen and cilia formation during zebrafish left-right organizer development.
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DOI：
10.1371/journal.pgen.1010765
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发表时间：
2023-05
期刊：
影响因子：
4.5
通讯作者：
中科院分区：
文献类型：
--
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在斑马鱼的左右组织者（Kupffer囊泡，KV）形成过程中的一个重要过程是通过发育延伸到KV腔中的KV细胞形成运动纤毛。KV管腔内活动纤毛的跳动引导流体流动以建立胚胎的左右轴。然而，KV细胞开始形成纤毛的时间点以及纤毛形成如何与KV管腔形成协调尚未研究。我们发现新生KV细胞在其中心体的随机细胞内位置形成纤毛，然后向含有囊性纤维化跨膜传导调节因子（CFTR）的形成顶端膜移动。使用光遗传学聚类方法，我们发现Rab35阳性膜招募Rab11来调节CFTR向顶膜的递送，这是管腔打开和随后纤毛延伸到管腔中所需的。一旦细胞内纤毛到达CFTR阳性顶膜，一旦管腔达到300 μm2的面积，Arl13b阳性纤毛以Rab8依赖性方式延伸并伸长到形成管腔中。这些研究表明，需要急性协调Rab8，Rab11和Rab35介导的膜运输事件，以确保适当的时间在KV发展过程中的管腔和纤毛形成。纤毛是从细胞表面延伸的基于微管的结构，以感知并将细胞外信号传递到细胞体。有缺陷的纤毛导致一系列的条件称为纤毛病。虽然纤毛形成的机制已在体外阐明，纤毛是如何在胚胎发育过程中在体内的研究较少。我们利用斑马鱼的器官的不对称性，也被称为库普弗囊泡（KV），作为一个模型来研究纤毛的形成在脊椎动物胚胎发育。在KV中，纤毛和管腔形成同时发生，使其成为研究这些过程如何协调的一个很好的模型。在这里，我们表明，KV前体细胞（背前体细胞）形成纤毛的细胞体积内延伸到腔的KV。为了确定在KV发展过程中纤毛形成的分子机制，我们专注于三个Rab GTP酶，Rab11，Rab8和Rab35，已牵连在纤毛或管腔形成独立的体外。我们使用光遗传学聚类与吗啉代耗竭方法并行显示，这三个Rab GTP酶协调特定的事件，在KV发展过程中控制纤毛和管腔形成。
An essential process during Danio rerio’s left-right organizer (Kupffer’s Vesicle, KV) formation is the formation of a motile cilium by developing KV cells which extends into the KV lumen. Beating of motile cilia within the KV lumen directs fluid flow to establish the embryo’s left-right axis. However, the timepoint at which KV cells start to form cilia and how cilia formation is coordinated with KV lumen formation have not been examined. We identified that nascent KV cells form cilia at their centrosomes at random intracellular positions that then move towards a forming apical membrane containing cystic fibrosis transmembrane conductance regulator (CFTR). Using optogenetic clustering approaches, we found that Rab35 positive membranes recruit Rab11 to modulate CFTR delivery to the apical membrane, which is required for lumen opening, and subsequent cilia extension into the lumen. Once the intracellular cilia reach the CFTR positive apical membrane, Arl13b-positive cilia extend and elongate in a Rab8 dependent manner into the forming lumen once the lumen reaches an area of 300 μm2. These studies demonstrate the need to acutely coordinate Rab8, Rab11, and Rab35-mediated membrane trafficking events to ensure appropriate timing in lumen and cilia formation during KV development. Cilia are microtubule-based structures that extend from the cell surface to sense and transmit extracellular cues to the cell body. Defective cilia result in a series of conditions known as ciliopathies. Although the mechanisms of cilia formation have been elucidated in vitro, how a cilium is made in vivo during embryogenesis is less studied. We take advantage of the Zebrafish’s organ of asymmetry, also known as the Kupffer’s Vesicle (KV), as a model to study cilia formation during vertebrate embryogenesis. In the KV, cilia and lumen formation occur concurrently, making it a great model to study how these processes are coordinated. Here we show that KV precursor cells (dorsal forerunner cells) form cilia within the cell volume before extending into the lumen of the KV. To determine the molecular mechanisms of cilia formation during KV development, we focus on three Rab GTPases, Rab11, Rab8 and Rab35, that have been implicated in cilia or lumen formation independently in vitro. We use optogenetic clustering in parallel with morpholino depletion approaches to show that these three Rab GTPases coordinate specific events that govern cilia and lumen formation during KV development.
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影响因子：
4.4
作者：
Krishnan N;Swoger M;Rathbun LI;Fioramonti PJ;Freshour J;Bates M;Patteson AE;Hehnly H
通讯作者：
Hehnly H
影响因子：
21.3
作者：
通讯作者：
--
影响因子：
5.5
作者：
Matsui T;Ishikawa H;Bessho Y
通讯作者：
Bessho Y
DOI：
10.1016/j.cub.2021.04.075
发表时间：
2021-07-12
期刊：
Current biology : CB
影响因子：
--
作者：
Ganga AK;Kennedy MC;Oguchi ME;Gray S;Oliver KE;Knight TA;De La Cruz EM;Homma Y;Fukuda M;Breslow DK
通讯作者：
Breslow DK
影响因子：
7.7
作者：
Kuhns, Stefanie;Seixas, Cecilia;Blacque, Oliver E.
通讯作者：
Blacque, Oliver E.