Assessment of ciliary phenotype in primary ciliary dyskinesia by micro-optical coherence tomography
George M. Solomon, … , Cecilia W. Lo, Steven M. Rowe
George M. Solomon, … , Cecilia W. Lo, Steven M. Rowe
Published March 9, 2017
Citation Information: JCI Insight. 2017;2(5):e91702. https://doi.org/10.1172/jci.insight.91702.
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Research Article Pulmonology

Assessment of ciliary phenotype in primary ciliary dyskinesia by micro-optical coherence tomography

Abstract

Ciliary motion defects cause defective mucociliary transport (MCT) in primary ciliary dyskinesia (PCD). Current diagnostic tests do not assess how MCT is affected by perturbation of ciliary motion. In this study, we sought to use micro-optical coherence tomography (μOCT) to delineate the mechanistic basis of cilia motion defects of PCD genes by functional categorization of cilia motion. Tracheae from three PCD mouse models were analyzed using μOCT to characterize ciliary motion and measure MCT. We developed multiple measures of ciliary activity, integrated these measures, and quantified dyskinesia by the angular range of the cilia effective stroke (ARC). Ccdc39–/– mice, with a known severe PCD mutation of ciliary axonemal organization, had absent motile ciliary regions, resulting in abrogated MCT. In contrast, Dnah5–/– mice, with a missense mutation of the outer dynein arms, had reduced ciliary beat frequency (CBF) but preserved motile area and ciliary stroke, maintaining some MCT. Wdr69–/– PCD mice exhibited normal motile area and CBF and partially delayed MCT due to abnormalities of ciliary ARC. Visualization of ciliary motion using μOCT provides quantitative assessment of ciliary motion and MCT. Comprehensive ciliary motion investigation in situ classifies ciliary motion defects and quantifies their contribution to delayed mucociliary clearance.

Authors

George M. Solomon, Richard Francis, Kengyeh K. Chu, Susan E. Birket, George Gabriel, John E. Trombley, Kristi L. Lemke, Nikolai Klena, Brett Turner, Guillermo J. Tearney, Cecilia W. Lo, Steven M. Rowe

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Figure 3

Characterization of Ccdc39–/– mice.

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Characterization of Ccdc39–/– mice. (A) Representative motile cilia area...
(A) Representative motile cilia area map from Ccdc39–/– tracheae at 5 days old indicates near absence of active ciliary movement in the mutant (n = 5). (B) Representative motile cilia area map from Ccdc39+/+ tracheae from a littermate control. Colored pixels indicate areas of active cilia movement at varied CBFs (n = 5). (C) Quantification of reduced percentage of motile ciliated areas in Ccdc39–/– mice. **P < 0.005. (D) Ciliary activity maps indicates undetectable (ND) CBFs in Ccdc39–/– mice. ****P < 0.0001. (E) ESR indicates that the reduced MCT in this mutant class is due to absent ciliary movement and reduced ciliary expression. (F) Residual motile cilia demonstrate dyskinesia with reduced arc angle in Ccdc39–/– mice. ****P < 0.0001. (G and H) Absent fluid transport is observed in Ccdc39–/– mice (G) compared with controls (H). (I) MCT is nearly absent in Ccdc39–/– mice due to effectively reduced ciliated area and movement. **P < 0.005. n = 6 Ccdc39+/+ animals and 5 Ccdc39–/– animals; 4 ROIs per trachea. All data are represented as mean (line) with minimum and maximum boundaries displayed. Statistical significance determined by Mann-Whitney t test. Scale bar: 10 microns (two dimensions, A and B); 10 micron by 10-second images (G and H).