@article{056cfb5d66cb4a09ac3a74afa2e1615a,
title = "Cryo-electron tomography reveals ciliary defects underlying human RSPH1 primary ciliary dyskinesia",
abstract = "Cilia play essential roles in normal human development and health; cilia dysfunction results in diseases such as primary ciliary dyskinesia (PCD). Despite their importance, the native structure of human cilia is unknown, and structural defects in the cilia of patients are often undetectable or remain elusive because of heterogeneity. Here we develop an approach that enables visualization of human (patient) cilia at high-resolution using cryo-electron tomography of samples obtained noninvasively by nasal scrape biopsy. We present the native 3D structures of normal and PCD-causing RSPH1-mutant human respiratory cilia in unprecedented detail; this allows comparisons of cilia structure across evolutionarily distant species and reveals the previously unknown primary defect and the heterogeneous secondary defects in RSPH1-mutant cilia. Our data provide evidence for structural and functional heterogeneity in radial spokes, suggest a mechanism for the milder RSPH1 PCD phenotype and demonstrate that cryo-electron tomography can be applied to human disease by directly imaging patient samples.",
author = "Jianfeng Lin and Weining Yin and Smith, {Maria C.} and Kangkang Song and Leigh, {Margaret W.} and Zariwala, {Maimoona A.} and Knowles, {Michael R.} and Ostrowski, {Lawrence E.} and Daniela Nicastro",
note = "Funding Information: We thank Daniel T.N. Chen (Brandeis University) for providing S. purpuratus sperm, Mary Porter (University of Minnesota) for providing the pseudo-wild-type Chlamydomonas strain and Chen Xu for providing EM training and management of the electron microscopes in the Louise Mashal Gabbay Cellular Visualization Facility. We thank David Mastronarde and John Heumann (University of Colorado at Boulder) for continued development of image processing tools, such as PEET classification. We also thank the UNC Cystic Fibrosis Cell and Tissue Culture Core (supported by R026-CR11 from the Cystic Fibrosis Foundation and NIH DK065988) for providing samples and expertise, and M. Coulouris for patient recruitment. This work was supported by funding from the National Institutes of Health (GM083122 to D.N., HL117836 to L.E.O., HL096458 and HL071798 to M.R.K.) and the March of Dimes Foundation (to D.N.) and by a private donation from Mr. Arnold Cohen (to D.N.). Publisher Copyright: {\textcopyright} 2014 Macmillan Publishers Limited. All rights reserved.",
year = "2014",
doi = "10.1038/ncomms6727",
language = "English (US)",
volume = "5",
journal = "Nature communications",
issn = "2041-1723",
publisher = "Nature Publishing Group",
}