Tppp3 determines basal body positioning and identity of respiratory cilia via microtubule assembly and sphingolipid homeostasis
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 ( Tppp3 ∆ex2-4/∆ex2-4 ; Tppp3 knockout (KO)) and found that the Tppp3 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 Tppp3 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 Tppp3 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.
Article Details
Journal Info
Proceedings of the National Academy of Sciences
National Academy of Sciences
Authors (29)
Takafumi Sakai
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology
Masahiro Kawakita
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology
Arashi Seki
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology
Katsuyoshi Takaoka
Graduate School of Frontier Biosciences, Osaka University
Misaki Kurata
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology
Junpei Torikai
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology
Sohshiroh Atsumi
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology
Kanji Yawata
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology
Kentaro Noi
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology
Yosuke Fukutani
Keiichi Noguchi
Instrumentation Analysis Center, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei-shi, Tokyo 184-8588, Japan
Hirofumi Kiyokawa
Laboratory for Lung Development and Regeneration, RIKEN Center for Biosystems Dynamics Research
Mitsuru Morimoto
Laboratory for Lung Development and Regeneration, RIKEN Center for Biosystems Dynamics Research
Eri Takeuchi
Department of Molecular Therapy, National Institute of Neuroscience, National Center of Neurology and Psychiatry
Katsura Minegishi
Department of Molecular Therapy, National Institute of Neuroscience, National Center of Neurology and Psychiatry
Yoshitsugu Aoki
Department of Molecular Therapy, National Institute of Neuroscience, National Center of Neurology and Psychiatry
Shoko Fujimura
Department of Physics, Gakushuin University
Takayuki Nishizaka
Department of Physics, Gakushuin University
Takanari Inoue
Department of Cell Biology, Johns Hopkins University School of Medicine
Kentaro Nagaoka
Department of Veterinary Medicine, Tokyo University of Agriculture and Technology
Hiroshi Tsugawa
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology
Hisayoshi Hayashi
School of Food and Nutritional Sciences, University of Shizuoka
Hiroshi Ishiguro
Department of Human Nutrition, Nagoya University Graduate School of Medicine
Takuma Kozono
Department of Applied Biological Science, Tokyo University of Agriculture and Technology
Ikuroh Ohsawa
Research Teams for Mechanism of Aging, Tokyo Metropolitan Institute of Gerontology
Yasunori Fujita
Research Teams for Mechanism of Aging, Tokyo Metropolitan Institute of Gerontology
Hiroaki Matsunami
Hiroshi Hamada
Graduate School of Frontier Biosciences, Osaka University
Kyosuke Shinohara
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology