Methods used for PCD diagnosis.
(A, B) Immunofluorescence co-staining of human respiratory epithelial cells with DNAH5-specific antibodies (red) and antibodies against acetylated α-tubulin (green). Nuclei were stained with Hoechst 33342 (blue). Overlays and bright-field images are shown on the right. Whereas in healthy human respiratory epithelial cells (wt, A) both DNAH5 and acetylated α-tubulin antibodies co-localize along the entire length of the ciliary axonemes, in an individual with an outer dynein arm defect (B), the ODA heavy chain DNAH5 is absent from the axonemes. (C) Transmission electron tomography of healthy respiratory epithelial cells (wt) showing no ultrastructural abnormality. Outer dynein arms (ODAs) are highlighted with red arrows. In an individual with DNAH5 mutations, ODAs are missing. (D) Diagram of ciliary beat patterns as deduced from high-speed videomicroscopy. A normal ciliary beat pattern (wt) is characterized by a strong beating stroke (symbolized in grey) followed by a recovery stroke (symbolized in green). In DNAH5 mutant cilia, only a minimal residual ciliary activity is present.