Tppp3 determines basal body positioning and identity of respiratory cilia via microtubule assembly and sphingolipid homeostasis

Cilia are hair-like organelles that protrude from the cell surface. In mammals, tracheal multiciliated cells (MCCs) play an important role in elimination of hazardous microorganisms by driving a unidirectional mucus flow. Although uniform orientation of ciliary beating is critical for the unidirectional flow, it remains unknown how MCCs establish uniform orientations and maintain identities of hundreds of ciliary membranes. This study focuses on investigating the roles of Tubulin Polymerization Promoting Family Member 3 (Tppp3) in MCC function. We generated a Tppp3-deficient mouse (<i>Tppp3</i><sup>∆ex2-4/∆ex2-4</sup>; <i>Tppp3</i> knockout (KO)) and found that the <i>Tppp3</i> KO mouse exhibited cough and hyposmia phenotype. The loss of Tppp3 disrupted the apical microtubules (MTs) meshwork in the tracheal MCCs, leading to random orientation and alignment of basal bodies (BBs) of the motile cilia. Unexpectedly, aberrant ciliary membrane fusions occurred in the trachea of the <i>Tppp3</i> KO mice. We examined the underlying molecular mechanism of the ciliary membrane fusion by isolating the tracheal cilium. Liquid Chromatography-Mass Spectrometry (LC-MS) analysis as well as pharmacological analysis revealed that hyperaccumulation of a long chain ceramide at the ciliary membrane caused the membrane fusion. In addition, sensory cilia formation was impaired in the olfactory sensory neuron of the <i>Tppp3</i> KO mice. Due to the lack of Tppp3, dendritic MT assembly that underlies long-range migration of BBs toward the cell surface was impaired. These findings demonstrate that Tppp3, as well as the defined intracellular MT architecture, regulate proper orientation/subcellular positioning of BBs and the independency of individual motile cilium membranes.

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Journal
Proceedings of the National Academy of Sciences
Published
2025-12-01
DOI
https://doi.org/10.1073/pnas.2503931122
Primary Topic
Cystic Fibrosis Research Advances
Type
article
Field-Weighted Citation Impact
0.00

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article

Tppp3 determines basal body positioning and identity of respiratory cilia via microtubule assembly and sphingolipid homeostasis

Kentaro Noi, Hiroshi Ishiguro, Yosuke Fukutani, Hiroshi Hamada et al.
Proceedings of the National Academy of Sciences
Cystic Fibrosis Research Advances
article

Tppp3 determines basal body positioning and identity of respiratory cilia via microtubule assembly and sphingolipid homeostasis

Kentaro Noi, Hiroshi Ishiguro, Yosuke Fukutani, Hiroshi Hamada, Keiichi Noguchi, Atsushi Seki, Katsura Minegishi, Ikuroh Ohsawa, Subaru Atsumi, Takanari Inoue, Yasunori Fujita, Takayuki Nishizaka, Kentaro Nagaoka, Hiroshi Tsugawa, Hirofumi Kiyokawa, Shoko Fujimura, Kyosuke Shinohara, K. Takaoka, Masahiro Kawakita, Yoshitsugu Aoki, Mitsuru Morimoto, Eri Takeuchi, Takuma Kozono, Hiroaki Matsunami, Hisayoshi Hayashi, Misaki Kurata, T Sakai, Junpei Torikai, Kanji Yawata
article en

Abstract

Cilia are hair-like organelles that protrude from the cell surface. In mammals, tracheal multiciliated cells (MCCs) play an important role in elimination of hazardous microorganisms by driving a unidirectional mucus flow. Although uniform orientation of ciliary beating is critical for the unidirectional flow, it remains unknown how MCCs establish uniform orientations and maintain identities of hundreds of ciliary membranes. This study focuses on investigating the roles of Tubulin Polymerization Promoting Family Member 3 (Tppp3) in MCC function. We generated a Tppp3-deficient mouse (<i>Tppp3</i><sup>∆ex2-4/∆ex2-4</sup>; <i>Tppp3</i> knockout (KO)) and found that the <i>Tppp3</i> KO mouse exhibited cough and hyposmia phenotype. The loss of Tppp3 disrupted the apical microtubules (MTs) meshwork in the tracheal MCCs, leading to random orientation and alignment of basal bodies (BBs) of the motile cilia. Unexpectedly, aberrant ciliary membrane fusions occurred in the trachea of the <i>Tppp3</i> KO mice. We examined the underlying molecular mechanism of the ciliary membrane fusion by isolating the tracheal cilium. Liquid Chromatography-Mass Spectrometry (LC-MS) analysis as well as pharmacological analysis revealed that hyperaccumulation of a long chain ceramide at the ciliary membrane caused the membrane fusion. In addition, sensory cilia formation was impaired in the olfactory sensory neuron of the <i>Tppp3</i> KO mice. Due to the lack of Tppp3, dendritic MT assembly that underlies long-range migration of BBs toward the cell surface was impaired. These findings demonstrate that Tppp3, as well as the defined intracellular MT architecture, regulate proper orientation/subcellular positioning of BBs and the independency of individual motile cilium membranes.

Proceedings of the National Academy of SciencesVol. 122(49)
Johns Hopkins University (US), University of Shizuoka (JP), Duke University (US), Johns Hopkins Medicine (US), Ube Frontier University (JP), National Center of Neurology and Psychiatry (JP), RIKEN Center for Biosystems Dynamics Research (JP), Tokyo Metropolitan Institute of Gerontology (JP), Food & Nutrition (NL), Institute of Neurobiology (BG), Gakushuin University (JP), Nagoya University (JP), Tokyo University of Agriculture and Technology (JP), The University of Osaka (JP)
Ministry of Education, Culture, Sports, Science and Technology, Tokyo University of Agriculture, Tokyo University of Agriculture and Technology, Japan Society for the Promotion of Science
Openalex Percentile: Top 44%
Cystic Fibrosis Research Advances
DOIOpen Access PDFOpenAlex
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