PIH1D3-knockout rats exhibit full ciliopathy features and dysfunctional pre-assembly and loading of dynein arms in motile cilia.
PIH1D3-knockout rats exhibit full ciliopathy features and dysfunctional pre-assembly and loading of dynein arms in motile cilia.
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DOI：
10.3389/fcell.2023.1282787
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发表时间：
2023
影响因子：
5.5
通讯作者：
中科院分区：
文献类型：
--
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背景：X连锁基因PIH 1域蛋白3（PIH 1D 3）的隐性突变导致家族性纤毛病变。PIH 1D 3缺陷与纤毛动力蛋白臂的缺陷有关，但PIH 1D 3具体如何影响动力蛋白臂的结构和功能尚不清楚。为了深入了解疾病的潜在机制，创建可靠的动物模型至关重要。在人类、大鼠和小鼠中，PIH 1D 3基因的一个拷贝位于X染色体上。有趣的是，小鼠在1号染色体上有一个额外的无内含子Pih 1d 3基因拷贝。为了开发准确的疾病模型，最好是操纵X连锁PIH 1D 3基因，该基因在内含子内含有用于精确基因表达的必需调控序列。本研究旨在开发一种定制的PIH 1D 3相关纤毛病变大鼠模型，最终目标是揭示该疾病中纤毛缺陷的复杂分子机制。 研究方法：通过在受精大鼠卵子内使用TALEN介导的非同源DNA重组创建了新型Pih 1d 3敲除（KO）大鼠，随后通过一系列行为和病理学测定进行了全面表征。进行了一系列的生化和组织学分析，以阐明与PIH 1D 3相互作用的蛋白质伴侣的身份，从而揭示了在这种情况下所涉及的复杂的分子机制。 结果如下：PIH 1D 3-KO大鼠再现了纤毛病变的主要特征，包括原位倒置、精母细胞存活和粘液纤毛清除缺陷以及围产期脑积水。我们揭示了PIH 1D 3在脑脊液循环中的新功能，并阐明了PIH 1D 3缺乏导致交通性脑积水的机制。PIH 1D 3与外动力蛋白臂（ODA）和内动力蛋白臂（IDA）的预组装和上传所需的蛋白质相互作用，调节纤毛中动力蛋白臂结构和功能的完整性。 结论：PIH 1D 3-KO大鼠真实地再现了PIH 1D 3缺乏相关纤毛病变的基本特征。PIH 1D 3与负责纤毛中动力蛋白臂的预组装和上传的蛋白质相互作用，并且其缺乏导致纤毛功能障碍，并且因此通过影响动力蛋白臂的预组装和上传而导致纤毛病。由此产生的大鼠模型是PIH 1D 3引起的疾病的机制研究的有价值的工具。
Background: Recessive mutation of the X-linked gene, PIH1 domain-containing protein 3 (PIH1D3), causes familial ciliopathy. PIH1D3 deficiency is associated with the defects of dynein arms in cilia, but how PIH1D3 specifically affects the structure and function of dynein arms is not understood yet. To gain insights into the underlying mechanisms of the disease, it is crucial to create a reliable animal model. In humans, rats, and mice, one copy of the PIH1D3 gene is located on the X chromosome. Interestingly, mice have an additional, intronless copy of the Pih1d3 gene on chromosome 1. To develop an accurate disease model, it is best to manipulate the X-linked PIH1D3 gene, which contains essential regulatory sequences within the introns for precise gene expression. This study aimed to develop a tailored rat model for PIH1D3-associated ciliopathy with the ultimate goal of uncovering the intricate molecular mechanisms responsible for ciliary defects in the disease. Methods: Novel Pih1d3-knockout (KO) rats were created by using TALEN-mediated non-homologous DNA recombination within fertilized rat eggs and, subsequently, underwent a comprehensive characterization through a battery of behavioral and pathological assays. A series of biochemical and histological analyses were conducted to elucidate the identity of protein partners that interact with PIH1D3, thus shedding light on the intricate molecular mechanisms involved in this context. Results: PIH1D3-KO rats reproduced the cardinal features of ciliopathy including situs inversus, defects in spermatocyte survival and mucociliary clearance, and perinatal hydrocephalus. We revealed the novel function of PIH1D3 in cerebrospinal fluid circulation and elucidated the mechanism by which PIH1D3 deficiency caused communicating hydrocephalus. PIH1D3 interacted with the proteins required for the pre-assembly and uploading of outer (ODA) and inner dynein arms (IDA), regulating the integrity of dynein arm structure and function in cilia. Conclusion: PIH1D3-KO rats faithfully reproduced the cardinal features of ciliopathy associated with PIH1D3 deficiency. PIH1D3 interacted with the proteins responsible for the pre-assembly and uploading of dynein arms in cilia, and its deficiency led to dysfunctional cilia and, thus, to ciliopathy by affecting the pre-assembly and uploading of dynein arms. The resultant rat model is a valuable tool for the mechanistic study of PIH1D3-caused diseases.
影响因子：
2.7
作者：
Wang R;Chen H;Wang X;Huang S;Xie A;Wu X
通讯作者：
Wu X
强烈截断的DNAAF4在果蝇纤毛动力蛋白组装中起着保守的作用，这是与DNAAF6的R2TP样伴侣复合物的一部分。
影响因子：
3.7
作者：
Lennon, Jennifer;zur Lage, Petra;von Kriegsheim, Alex;Jarman, Andrew P.
通讯作者：
Jarman, Andrew P.