Laboratory for Cell Adhesion and Tissue Patterning.
Division of Embryology, National Institute for Basic Biology, and Department of Basic Biology, School of Life Science, SOKENDAI, the Graduate University for Advanced Studies, Okazaki, 444-8787 Japan.
Laboratory for Organismal Patterning.
Nonequilibrium Physics of Living Matter RIKEN Hakubi Research Team.
Laboratory for Physical Biology, and.
Ultrastructural Research Team, RIKEN Center for Life Science Technologies, Kobe 650-0047, Japan.
Laboratory for Ultrastructural Research, RIKEN Center for Biosystems Dynamics Research, Kobe 650-0047, Japan.
Laboratory of Cytoskeletal Logistics, Center for Advanced Biomedical Sciences (TWIns), Waseda University, Tokyo 162-8480, Japan.
Waseda Institute for Advanced Study, Waseda University, Tokyo 169-0051, Japan.
Major in Bioscience, Global Center for Science and Engineering, Faculty of Science and Engineering, Waseda University, Tokyo 169-8555, Japan.
Motile cilia of multiciliated epithelial cells undergo synchronized beating to produce fluid flow along the luminal surface of various organs. Each motile cilium consists of an axoneme and a basal body, which are linked by a ‘transition zone’. The axoneme exhibits a characteristic 9+2 microtubule arrangement important for ciliary motion, but how this microtubule system is generated is not yet fully understood. Here we show that CAMSAP3, a protein that can stabilize the minus end of a microtubule, concentrates at multiple sites of the cilium–basal body complex, including the upper region of the transition zone or the axonemal basal plate where the central pair of microtubules (CP) initiates. CAMSAP3 dysfunction resulted in loss of the CP and partial distortion of the basal plate, as well as the failure of multicilia to undergo synchronized beating. These findings suggest that CAMSAP3 plays pivotal roles in the formation or stabilization of the CP by localizing at the basal region of the axoneme, and thereby supports the coordinated motion of multicilia in airway epithelial cells.
Audio S1: Respiratory sound of a P50 Camsap3 mutant mouse.Download Original Video (.8 MB)https://ascb-prod-streaming.literatumonline.com/journals/content/mboc/0/mboc.ahead-of-print/mbc.e21-06-0303/20210726/media/mc-e21-06-0303-s01.,900,768,652,642,.mp4.m3u8?b92b4ad1b4f274c7087751851dabb28b5eff7736ed2d13488ba0685b5d4b43f735f6b48a41f0861e362653b46f872d70e7a199ee5ab054e5562a0a61d56adcc26fec2da2c8c4e10d496af1d512ad07ae23104960c1d1c1007580c2690b66824d6b6222ec22ca50156bfd6634651a5d96e0a57c2906bdfb19e8b75356a5083d984d73f34e2b591ff9e4fca206d4f8366e49e93843c2440b9af95650023ad95ab608e03ba8b2ec1951e3Movie S1: Beating multicilia in a wild-type airway epithelial cell. The time-lapse images were acquired at 300 fps and displayed at 31 fps.Download Original Video (1.0 MB)https://ascb-prod-streaming.literatumonline.com/journals/content/mboc/0/mboc.ahead-of-print/mbc.e21-06-0303/20210726/media/mc-e21-06-0303-s02.,652,642,.mp4.m3u8?b92b4ad1b4f274c70877518515abb28bda92fbabe7b929571bd415190bf44d1790b7324fb90f184f91fab10e395327a1bef9725d0c99a358bbbe69c082a95f9d49957eb2932b2be9a6f328f5ec3ab2a0e23b009e247a8c8af6d8f02951b492f63ab95f3990c432d488249842bce4d11f4f0801c17519cdcd55fad1600e6c23d5f12d0228324ef46da5c7ca80b447d4f5eca2723e10989407b535bd534ce60b8bbfMovie S2: Beating multicilia in a Camsap3-mutated airway epithelial cell. The time-lapse images were acquired at 300 fps and displayed at 31 fps.Download Original Video (1.0 MB)https://ascb-prod-streaming.literatumonline.com/journals/content/mboc/0/mboc.ahead-of-print/mbc.e21-06-0303/20210726/media/mc-e21-06-0303-s03.,652,642,.mp4.m3u8?b92b4ad1b4f274c70877518515abb28bda92fbabe7b929571bd415190bf44d1790b7324fb90f184f91fab10e395327a1bef9725d0c99a358bbbe69c082a95f9d49957eb2932b2be9a6f328f5ec3ab2a0e23b009e247a8c8af6d8f02951b492f63ab95f3990c432d488249842bce4d11f4f0801c17519cdcd54fad1600e6c23d5ac68e5fdf756a8854a908ce4ed78d7081c9e597b7d2921432d04fa4a529a474250Movie S3: Beating multicilia in a wild-type airway epithelial cell, which is overlaid with a flow vector field calculated by PIV analysis. The flow vector field was also shown separately at the right. The scale arrow and scale bar are 20 μm/s and 1 m, respectively. Part of Video S1 was used for this analysis.Download Original Video (2.7 MB)https://ascb-prod-streaming.literatumonline.com/journals/content/mboc/0/mboc.ahead-of-print/mbc.e21-06-0303/20210726/media/mc-e21-06-0303-s04.,1200,960,900,768,652,642,.mp4.m3u8?b92b4ad1b4f274c7087751841cabb28b19a3e23570e4794628d00a388fabd4f47061f4b4b81c11dac4c336cd5be6a2a6a495e5def8479aeedf32e6f4ec8f325c8354b92f9a15e63bba6a844857b79b5251eb2f2c474ab98260698f38258eb6f58f90a295269167ddbe8c19f299bf0805081cca0b9e256ecf7297e3f8215dc03fb2e0121299a0675f1d9f8609b58d044bc1b68f31cdf1ee6dad0e430a9ebb0557a18ad34ab01fadf8a6974c0059b42a69759bMovie S4: Beating multicilia in a Camsap3-mutated airway epithelial cell, which is overlaid with a flow vector field calculated by PIV analysis. The flow vector field was also shown separately at the right. The scale arrow and scale bar are 20 μm/s and 1 μm, respectively. Part of Video S2 was used for this analysis.Download Original Video (2.8 MB)https://ascb-prod-streaming.literatumonline.com/journals/content/mboc/0/mboc.ahead-of-print/mbc.e21-06-0303/20210726/media/mc-e21-06-0303-s05.,1200,960,900,768,652,642,.mp4.m3u8?b92b4ad1b4f274c7087751841cabb28b19a3e23570e4794628d00a388fabd4f47061f4b4b81c11dac4c336cd5be6a2a6a495e5def8479aeedf32e6f4ec8f325c8354b92f9a15e63bba6a844857b79b5251eb2f2c474ab98260698f38258eb6f58f90a295269167ddbe8c19f299bf0805081cca0b9e256ecf7397e3f8215dc03f9679bb9bcb8b2119749ee745b79cc7be86c76663f2243a2b32b0a241bdf1b26db05a3326f602117c41f4f79534b31de3a66fMovie S5: Animation of sequential optical sections, each of which is 0.2 m thick, of a wild-type multi-ciliated epithelial cell, in which GFP-Centrin (green) and -tubulin (magenta) are visualized by fluorescence signals. The animation view begins around the level where the array of GFP-Centrin is detectable, then shifts toward a more basal view of the cell at a speed of 2 fps. See also Figure 6A.Download Original Video (.2 MB)https://ascb-prod-streaming.literatumonline.com/journals/content/mboc/0/mboc.ahead-of-print/mbc.e21-06-0303/20210726/media/mc-e21-06-0303-s06.,652,642,.mp4.m3u8?b92b4ad1b4f274c70877518515abb28bda92fbabe7b929571bd415190bf44d1790b7324fb90f184f91fab10e395327a1bef9725d0c99a358bbbe69c082a95f9d49957eb2932b2be9a6f328f5ec3ab2a0e23b009e247a8c8af6d8f02951b492f63ab95f3990c432d488249842bce4d11f4f0801c17519cdcd51fad1600e6c23d5a91d300a2f17a0915fb1d06915b77411b12a59719095a792f907c352144b6974e8Movie S6: Animation of sequential optical sections, each of which is 0.2 m thick, of a Camsap3-mutated multi-ciliated epithelial cell, in which GFP-Centrin (green) and -tubulin (magenta) are visualized by fluorescence signals. The animation view begins at the level where the array of GFP-Centrin is detectable, then shifts toward a more basal view of the cell at a speed of 2 fps. See also Figure 6B.Download Original Video (.2 MB)https://ascb-prod-streaming.literatumonline.com/journals/content/mboc/0/mboc.ahead-of-print/mbc.e21-06-0303/20210726/media/mc-e21-06-0303-s07.,652,642,.mp4.m3u8?b92b4ad1b4f274c70877518515abb28bda92fbabe7b929571bd415190bf44d1790b7324fb90f184f91fab10e395327a1bef9725d0c99a358bbbe69c082a95f9d49957eb2932b2be9a6f328f5ec3ab2a0e23b009e247a8c8af6d8f02951b492f63ab95f3990c432d488249842bce4d11f4f0801c17519cdcd50fad1600e6c23d5a87a3ab234971ddf823bd6378e639c4b5e0573b31c4cca370bfa2c01a4be2e9191