Yap Yi Tian, Pan Jiehong, Xu Jian, Yuan Shuo, Niu Changmin, Zheng Cheng, Li Wei, Zhou Ting, Li Tao, Zhang Yong, Holtzman Michael J, Pazour Gregory J, Hess Rex A, Kelly Christopher V, Touré Aminata, Brody Steven L, Zhang Zhibing
Department of Physiology, Wayne State University, 275 E Hancock Street, Detroit, MI, 48201, USA.
Division of Pulmonary and Critical Care Medicine, Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA.
Cell Mol Life Sci. 2025 May 10;82(1):198. doi: 10.1007/s00018-025-05710-z.
Cilia are microtubular structures extending from the surface of most mammalian cells. They can be categorized as motile cilia and primary sensory cilia. Both types possess intraflagellar transport (IFT) machinery, composed of unique protein complexes that travel along the microtubules to deliver proteins for ciliary and flagellar assembly, disassembly, and homeostasis. Although the role of IFT in primary cilia formation has been well studied, little is known about its role in mammalian motile cilia assembly. We generated conditional knockout mice by breeding floxed Ift140 mice with the FOXJ1-Cre transgenic mouse line to specifically delete Ift140 from cells that assemble motile cilia. Mice with Ift140 deficiency did not have laterality defects or gross; however most died prior to sexual maturity. Those mutants that survived to adulthood were completely infertile. Males demonstrated abnormal spermatogenesis associated with reduced sperm count and motility, together with short length flagella, and abnormal morphology. Cilia length was diminished in the epithelial cells of the efferent ductules and airways. Cilia from cultured tracheal epithelial cells were also short and had reduced beat frequency (CBF). Ultrastructural studies revealed the presence of inner and outer dynein arms, but an abnormal central apparatus, and the accumulation of particles within the cilia. Overall, the short length and abnormal localization of ciliary proteins in Ift140 conditional mutants resulted in inadequate cilia function despite proper localization of the dynein motor complexes. We propose a key role of Ift140 for motile cilia assembly in certain tissues and suggest that genetic alterations of IFT140 could be associated with motile ciliopathies.
纤毛是从大多数哺乳动物细胞表面伸出的微管结构。它们可分为运动纤毛和初级感觉纤毛。这两种类型都拥有鞭毛内运输(IFT)机制,该机制由独特的蛋白质复合物组成,这些复合物沿着微管移动,为纤毛和鞭毛的组装、拆卸及体内平衡输送蛋白质。尽管IFT在初级纤毛形成中的作用已得到充分研究,但对其在哺乳动物运动纤毛组装中的作用却知之甚少。我们通过将携带floxed Ift140的小鼠与FOXJ1-Cre转基因小鼠品系杂交,生成条件性敲除小鼠,以特异性地从组装运动纤毛的细胞中删除Ift140。Ift140缺陷的小鼠没有偏侧性缺陷或大体异常;然而,大多数在性成熟前死亡。那些存活到成年的突变体完全不育。雄性表现出与精子数量减少、活力降低相关的异常精子发生,同时鞭毛长度缩短且形态异常。输出小管和气道上皮细胞中的纤毛长度缩短。培养的气管上皮细胞的纤毛也很短,且摆动频率(CBF)降低。超微结构研究显示存在内、外动力蛋白臂,但中央装置异常,且纤毛内有颗粒堆积。总体而言,尽管动力蛋白运动复合物定位正常,但Ift140条件性突变体中纤毛蛋白的短长度和异常定位导致纤毛功能不足。我们提出Ift140在某些组织的运动纤毛组装中起关键作用,并表明IFT140的基因改变可能与运动性纤毛病有关。