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.
authors.affiliation: Rosenstiel Basic Medical Sciences Research Center, Department of Biology, Brandeis University, Waltham, Massachusetts 02454, USA.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Lin
authors.affiliation: Cystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
authors.forename: Weining
authors.initials: W
authors.lastname: Yin
authors.affiliation: Rosenstiel Basic Medical Sciences Research Center, Department of Biology, Brandeis University, Waltham, Massachusetts 02454, USA.
authors.forename: Maria C
authors.initials: MC
authors.lastname: Smith
authors.affiliation: Rosenstiel Basic Medical Sciences Research Center, Department of Biology, Brandeis University, Waltham, Massachusetts 02454, USA.
authors.forename: Kangkang
authors.initials: K
authors.lastname: Song
authors.affiliation: Department of Pediatrics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599, USA.
authors.forename: Margaret W
authors.initials: MW
authors.lastname: Leigh
authors.affiliation: Department of Pathology and Laboratory Medicine, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599, USA.
authors.forename: Maimoona A
authors.initials: MA
authors.lastname: Zariwala
authors.affiliation: Department of Medicine, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599, USA.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Knowles
authors.affiliation: Cystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
authors.forename: Lawrence E
authors.initials: LE
authors.lastname: Ostrowski
authors.affiliation: Rosenstiel Basic Medical Sciences Research Center, Department of Biology, Brandeis University, Waltham, Massachusetts 02454, USA.
authors.forename: Daniela
authors.initials: D
authors.lastname: Nicastro
mesh_terms: D002923:Cilia / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure*; D020285:Cryoelectron Microscopy / Q000379:methods*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D055032:Electron Microscope Tomography / Q000379:methods*; D006801:Humans; D007619:Kartagener Syndrome / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; C484746:RSPH1 protein, human