TY - JOUR
T1 - CAMSAP3 facilitates basal body polarity and the formation of the central pair of microtubules in motile cilia
AU - Robinson, Alan M.
AU - Takahashi, Satoe
AU - Brotslaw, Eva J.
AU - Ahmad, Aisha
AU - Ferrer, Emma
AU - Procissi, Daniele
AU - Richter, Claus Peter
AU - Cheatham, Mary Ann
AU - Mitchell, Brian J.
AU - Zheng, Jing
N1 - Funding Information:
ACKNOWLEDGMENTS. We thank Drs. Jonathan Siegel, Donna Whitlon (North-western University), and Masatoshi Takeichi (RIKEN Center for Developmental Biology) for providing instruments and reagents, and Mr. Taewoo Kim (North-western University) for participating in the data analysis. Imaging work was performed at the Northwestern University Center for Advanced Microscopy, which is supported by National Cancer Institute (NCI) grant CCSG P30 CA060553 and National Institutes of Health NIH grant 1S10OD016342-01. MRI and CT scans were conducted at the Center for Translational Imaging Small Animal Imaging Laboratory. Rederivation of Camsap3tm1a lines was performed by the Transgenic and Targeted Mutagenesis Laboratory of Northwestern University. This research was funded by the Knowles Hearing Center (to J.Z.) and by National Institute on Deafness and Other Communication Disorders (NIDCD) grant R56 DC011813 (to J.Z.), grant DC000089 (to M.A.C.), and grant GM089970 (to B.J.M.).
Funding Information:
We thank Drs. Jonathan Siegel, Donna Whitlon (Northwestern University), and Masatoshi Takeichi (RIKEN Center for Developmental Biology) for providing instruments and reagents, and Mr. Taewoo Kim (Northwestern University) for participating in the data analysis. Imaging work was performed at the Northwestern University Center for Advanced Microscopy, which is supported by National Cancer Institute (NCI) grant CCSG P30 CA060553 and National Institutes of Health NIH grant 1S10OD016342-01. MRI and CT scans were conducted at the Center for Translational Imaging Small Animal Imaging Laboratory. Rederivation of Camsap3tm1a lines was performed by the Transgenic and Targeted Mutagenesis Laboratory of Northwestern University. This research was funded by the Knowles Hearing Center (to J.Z.) and by National Institute on Deafness and Other Communication Disorders (NIDCD) grant R56 DC011813 (to J.Z.), grant DC000089 (to M.A.C.), and grant GM089970 (to B.J.M.).
Publisher Copyright:
© 2020 National Academy of Sciences. All rights reserved.
PY - 2020/6/16
Y1 - 2020/6/16
N2 - Synchronized beating of cilia on multiciliated cells (MCCs) generates a directional flow of mucus across epithelia. This motility requires a "9 + 2" microtubule (MT) configuration in axonemes and the unidirectional array of basal bodies of cilia on the MCCs. However, it is not fully understood what components are needed for central MT-pair assembly as they are not continuous with basal bodies in contrast to the nine outer MT doublets. In this study, we discovered that a homozygous knockdown mouse model for MT minus-end regulator calmodulin-regulated spectrin-associated protein 3 (CAMSAP3), Camsap3tm1a/tm1a, exhibited multiple phenotypes, some of which are typical of primary ciliary dyskinesia (PCD), a condition caused by motile cilia defects. Anatomical examination of Camsap3tm1a/tm1a mice revealed severe nasal airway blockage and abnormal ciliary morphologies in nasal MCCs. MCCs from different tissues exhibited defective synchronized beating and ineffective generation of directional flow likely underlying the PCD-like phenotypes. In normal mice, CAMSAP3 localized to the base of axonemes and at the basal bodies in MCCs. However, in Camsap3tm1a/tm1a, MCCs lacked CAMSAP3 at the ciliary base. Importantly, the central MT pairs were missing in the majority of cilia, and the polarity of the basal bodies was disorganized. These phenotypes were further confirmed in MCCs of Xenopus embryos when CAMSAP3 expression was knocked down by morpholino injection. Taken together, we identified CAMSAP3 as being important for the formation of central MT pairs, proper orientation of basal bodies, and synchronized beating of motile cilia.
AB - Synchronized beating of cilia on multiciliated cells (MCCs) generates a directional flow of mucus across epithelia. This motility requires a "9 + 2" microtubule (MT) configuration in axonemes and the unidirectional array of basal bodies of cilia on the MCCs. However, it is not fully understood what components are needed for central MT-pair assembly as they are not continuous with basal bodies in contrast to the nine outer MT doublets. In this study, we discovered that a homozygous knockdown mouse model for MT minus-end regulator calmodulin-regulated spectrin-associated protein 3 (CAMSAP3), Camsap3tm1a/tm1a, exhibited multiple phenotypes, some of which are typical of primary ciliary dyskinesia (PCD), a condition caused by motile cilia defects. Anatomical examination of Camsap3tm1a/tm1a mice revealed severe nasal airway blockage and abnormal ciliary morphologies in nasal MCCs. MCCs from different tissues exhibited defective synchronized beating and ineffective generation of directional flow likely underlying the PCD-like phenotypes. In normal mice, CAMSAP3 localized to the base of axonemes and at the basal bodies in MCCs. However, in Camsap3tm1a/tm1a, MCCs lacked CAMSAP3 at the ciliary base. Importantly, the central MT pairs were missing in the majority of cilia, and the polarity of the basal bodies was disorganized. These phenotypes were further confirmed in MCCs of Xenopus embryos when CAMSAP3 expression was knocked down by morpholino injection. Taken together, we identified CAMSAP3 as being important for the formation of central MT pairs, proper orientation of basal bodies, and synchronized beating of motile cilia.
KW - Basal body orientation
KW - CAMSAP3
KW - Central MT pair
KW - Motile cilia
KW - Primary ciliary dyskinesia
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U2 - 10.1073/pnas.1907335117
DO - 10.1073/pnas.1907335117
M3 - Article
C2 - 32482850
AN - SCOPUS:85086681858
SN - 0027-8424
VL - 117
SP - 13571
EP - 13579
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 24
ER -