Global genetic analysis in mice unveils central role for cilia in congenital heart disease.

Congenital heart disease (CHD) is the most prevalent birth defect, affecting nearly 1% of live births; the incidence of CHD is up to tenfold higher in human fetuses. A genetic contribution is strongly suggested by the association of CHD with chromosome abnormalities and high recurrence risk. Here we report findings from a recessive forward genetic screen in fetal mice, showing that cilia and cilia-transduced cell signalling have important roles in the pathogenesis of CHD. The cilium is an evolutionarily conserved organelle projecting from the cell surface with essential roles in diverse cellular processes. Using echocardiography, we ultrasound scanned 87,355 chemically mutagenized C57BL/6J fetal mice and recovered 218 CHD mouse models. Whole-exome sequencing identified 91 recessive CHD mutations in 61 genes. This included 34 cilia-related genes, 16 genes involved in cilia-transduced cell signalling, and 10 genes regulating vesicular trafficking, a pathway important for ciliogenesis and cell signalling. Surprisingly, many CHD genes encoded interacting proteins, suggesting that an interactome protein network may provide a larger genomic context for CHD pathogenesis. These findings provide novel insights into the potential Mendelian genetic contribution to CHD in the fetal population, a segment of the human population not well studied. We note that the pathways identified show overlap with CHD candidate genes recovered in CHD patients, suggesting that they may have relevance to the more complex genetics of CHD overall. These CHD mouse models and >8,000 incidental mutations have been sperm archived, creating a rich public resource for human disease modelling.

The tubulin deglutamylase CCPP-1 regulates the function and stability of sensory cilia in C. elegans.

BACKGROUND: Posttranslational modifications (PTMs) such as acetylation, detyrosination, and polyglutamylation have long been considered markers of stable microtubules and have recently been proposed to guide molecular motors to specific subcellular destinations. Microtubules can be deglutamylated by the cytosolic carboxypeptidase CCP1. Loss of CCP1 in mice causes cerebellar Purkinje cell degeneration. Cilia, which are conserved organelles that play important diverse roles in animal development and sensation, contain axonemes comprising microtubules that are especially prone to PTMs. RESULTS: Here, we report that a CCP1 homolog, CCPP-1, regulates the ciliary localization of the kinesin-3 KLP-6 and the polycystin PKD-2 in male-specific sensory neurons in C. elegans. In male-specific CEM (cephalic sensilla, male) cilia, ccpp-1 also controls the velocity of the kinesin-2 OSM-3/KIF17 without affecting the transport of kinesin-II cargo. In the core ciliated nervous system of both males and hermaphrodites, loss of ccpp-1 causes progressive defects in amphid and phasmid sensory cilia, suggesting that CCPP-1 activity is required for ciliary maintenance but not ciliogenesis. Affected cilia exhibit defective B-tubules. Loss of TTLL-4, a polyglutamylating enzyme of the tubulin tyrosine ligase-like family, suppresses progressive ciliary defects in ccpp-1 mutants. CONCLUSIONS: Our studies suggest that CCPP-1 acts as a tubulin deglutamylase that regulates the localization and velocity of kinesin motors and the structural integrity of microtubules in sensory cilia of a multicellular, living animal. We propose that the neuronal degeneration caused by loss of CCP1 in mammals may represent a novel ciliopathy in which cilia are formed but not maintained, depriving the cell of cilia-based signal transduction.

[Evolution of the tubotympanal mucociliary system activity evaluated with scintigraphy, after ventilation tube insertion in adults with chronic otitis media with effusion]. / Evolución de la actividad mucociliar tubotimpánica, estudiada por gammagrafía, tras la colocación de tubos de ventilación en adultos con otitis media seromucosa crónica.

INTRODUCTION AND GOALS: Tubotympanal mucociliary function is damaged in patients with otitis media with effusion. Our objective was to study its situation and recovery after timopanostomy tube insertion. METHODS: We evaluated the mucociliary activity of the ear and Eustachian tube with scintigraphy in four groups of adults with chronic otitis media with effusion after ventilation tube insertion. The test was performed on 3 days (Group I), 6 days (Group II), 9 days (Group III) and 12 days (Group IV) after tympanostomy tube insertion. RESULTS: After excluding some patients with limited otorrhea, the percentages of patients that normalised mucociliary function in the different groups were: Group I, 0 of 13, 0%; Group II, 4 of 15 patients, 26,6%; Group III, 14 de 18 cases, 88,8%; and, in Group IV, 18 of 18, 100%. CONCLUSIONS: Mucociliary function is altered in patients with chronic otitis media with effusion. With the insertion of tympanostomy tubes, mucociliary clearance is restored progressively, reaching normal after 12 days in all the patients.

Obstructive azoospermia associated with chronic sinopulmonary infection and situs inversus totalis.

We describe 2 cases of obstructive azoospermia associated with situs inversus and sinopulmonary infection due to ciliary defects. Electron microscopy of testicular sperm flagella demonstrated normal morphology with nine peripheral doublets surrounding a central pair and complete sets of inner and outer dynein arms. Electron microscopy of the nasal mucosa revealed partial defects of the dynein arms of cilia, although the "9+2" morphology was preserved. Our cases were considered unique variants of Young's syndrome but also had characteristic features of Kartagener syndrome, and thus support the hypothesis that Young's syndrome has a genetic etiology similar to that of Kartagener syndrome.

The reproductive significance of human Fallopian tube cilia.

Effective tubal transport of ova, sperm and embryos is a prerequisite for successful spontaneous pregnancy. Although there is much yet to be discovered about the mechanisms involved, it is evident that tubal transit is a far more complicated process than initially thought. Propulsion of gametes and embryos is achieved by complex interaction between muscle contractions, ciliary activity and the flow of tubal secretions. Evidence is accumulating of the important and possibly pre-eminent role of ciliary motion in this process; and this review describes current knowledge about ciliary activity and its physiological regulation. There is also a description of the effects on ciliary function of cigarette smoking and various pathological states, including endometriosis and microbial infection, with consideration given as to how altered ciliary activity may impact upon fertility.

A comparative study between ciliary count and the degree of opacity of paranasal sinus CT scans in chronic sinusitis pre and post FESS.

OBJECTIVE: To investigate the relationship between the extent of sinus disease in chronic sinusitis as detected radiologically by computed tomography (CT) scan and the population of cilia (ciliary area) both before and after functional endoscopic sinus surgery (FESS). In a simple way this is a trial to statistically prove that the CT scan could be a valid indicator and a mirror of the histological status of the sinus mucosa. DESIGN: Twenty adult patients were enrolled in this study. Radiological extension of the sinus disease was quantitated using the classification proposed by Kennedy in 1992 and the ciliary population was studied using scanning electron microscopy and image analysis softwares. RESULTS AND CONCLUSION: The more advanced the sinusitis, as evidenced by CT scans, the more the expected reduction in the ciliary area (CA) and in the ciliary count. But after FESS the degree of ciliary regeneration does not depend statistically on the radiological condition of the sinuses and the degree of opacity prior to intervention, i.e. a statistically valid inverse relationship exists between the radiological stage of sinusitis and the ciliary population pre-operatively but the same relation does not extend to the ciliary population post-operatively.

Nasal mucociliary transport: new evidence for a key role of ciliary beat frequency.

OBJECTIVES/HYPOTHESIS: Mucociliary transport is an important defense mechanism of the respiratory tract. Nonetheless, the factors determining mucociliary transport are only partially understood. Ciliary beat frequency is assumed to be one of the main parameters, although the experimental evidence remains inconclusive. STUDY DESIGN: Comparing influences on mucociliary transport to influences on ciliary beat frequency. METHODS: The present study measures the effects on mucociliary transport of two ciliary beat frequency-inhibiting compounds (0.1% xylometazoline and 0.9% NaCl) and a ciliary beat frequency enhancer (0.1% salbutamol). The measurements were performed by a technetium-99m nebulizing scintigraphic method. The experiments were carried out in 15 healthy young volunteers. RESULTS: The 0.1% xylometazoline appeared to slow ciliary transport, although the decrease was not significant (P = .44). The 0.9% NaCl did reduce mucociliary transport significantly (P = .033). The 0.1% salbutamol resulted in a highly significant increase of mucociliary transport (P = .009). Xylometazoline brings about drastic changes in the nasal cavity, both anatomically and physiologically. Any comparison of mucociliary transport before and after using this vasoconstrictive agent must take this effect into account. CONCLUSIONS: The present study demonstrates a significant similarity in the effects of NaCl and salbutamol on ciliary beat frequency in vitro and on mucociliary transport in vivo. The evidence from our experiments suggests that ciliary beat frequency is a determining factor in the mucociliary transport rate in the nose.

Cisplatin induces modifications in the development of cell surface patterns of ciliates.

Cisplatin [cis-dichlorodiammineplatinum (II)] brings about significant quantitative modifications in the development of cell surface patterns in two unrelated ciliates: Stylonychia and Tetrahymena. Cells cultured in the presence of cisplatin exhibit the formation of supernumerary surface structures in the form of extra cilia/cirri (fused cilia) and other organized ciliary organelles. The metal-induced formation of extra primordia and their differentiation into supernumerary ciliary structures is governed by the normal rules of development. Additional structures are accommodated within the framework of a defined pattern, suggesting the existence of an overall global control of pattern formation. The modified pattern is rectified to its normal state during post-treatment fission cycles, suggesting the role of the cell membrane in correcting developmental errors.

[Comparative studies of ciliary epithelial morphological and radiologic findings in the osteomeatal complex].

In this study, we evaluated quantitatively preoperative coronal CT scan in 30 nasal cavities of 22 patients with chronic sinusitis who underwent functional endoscopic sinus surgery. The ciliated area of the osteomeatal complex (OMC) from these patients was quantitatively observed by scanning electron microscope and image analysis. Our results showed a positive correlation between ciliary epithelial pathological changes in OMC and disease degree of this area in CT scan.

Morphologic alterations in the cilia of a cat.

Based upon ultrastructural findings and computed tomography, a presumptive diagnosis of feline primary ciliary dyskinesia was made in a 2.5-year-old cat. The cat demonstrated morphologic alterations in the ultrastructure of oviductal cilia. In the oviduct, axonemal abnormalities were detected in 132 (20%) of 660 cross-sectioned cilia. The main ultrastructural ciliary defects were a lack of central microtubules, transposition of peripheral doublets to the center of the axoneme, supernumerary microtubules, and morphologic abnormalities of peripheral doublets. Computed tomography of the chest was consistent with early lesions of bronchiectasis, suggesting chronic stagnation of respiratory secretions, attributed to abnormal function of respiratory cilia. Specifically, the cranial branches of the cat's bronchi were wider and thicker than those of five healthy controls. Foci of pleural thickening and interstitial enlargement were also observed.

Immotile cilia syndrome: nasal mucociliary function and nasal ciliary abnormalities.

We present 17 patients with a typical symptomatology of immotile cilia syndrome, seven of them with complete situs inversus. Firstly, a study of the nasal mucociliary transport was made by means of the radioisotopic technique with serum albumin-Tc99m. In all cases there was absence of transport. Secondly, we studied the ultrastructure of the nasal cilia. Defects in the dynein arms were frequently found (65%). In two cases (11%) there were no cilia; in two other cases the cilia were normal, and in another two cases alterations of the central pair of microtubules were seen. Ciliary complexes were detected in all cases. We conclude that in patients with chronic or recurrent infections of the airways without known cause we must initiate a study of the nasal mucociliary transport. If this is absent or decreased, study of ciliary ultrastructure should be carried out. If mucociliary transport is normal, immotile cilia syndrome is ruled out and ultrastructural study of the cilia is not required.

Sugar-binding sites with specificity to N-acetyl-D-glucosaminides in middle ear mucosa of the guinea pig.

The distribution of sugar-binding sites was analyzed in Lowicryl K4M-embedded guinea pig middle ear mucosa. Four neoglycoproteins and a glycoprotein were used as probes: N-acetyl-D-glucosamine (GlcNAc), D-mannose, N-acetyl-D-galactosamine, or L-fucose carrying bovine serum albumin (BSA) and asialofetuin with terminal D-galactosyl sugar residues. Each probe was then labelled with 15 nm colloidal gold. Incubation of ultrathin sections with gold-labelled p-aminophenyl N-acetyl-beta-D-glucosaminide-BSA (GlcNAc/BSA/gold) led to binding on mucosal cilial, microvilli, rough endoplasmic reticulum, mitochondria, and nuclei. No binding occurred with asialofetuin or neoglycoproteins containing mannose, N-acetylgalactosamine or fucose. Various control experiments showed that specificity of GlcNAc/BSA/gold binding was directed towards N-acetylglucosaminyl residues expressed on the neoglycoprotein. Competitive sugar inhibition with GlcNAc and its derivatives suggested that the strong affinity for GlcNAc-binding sites took place in a complex formation with sugar residues bound to a carrier protein. The existence of a hydrophobic region close to the sugar-binding site was also suggested.

[Radionuclide evaluation of the mucociliary function of the mucosa of the nasal cavity in chronic lung diseases]. / Radionuklidnaia otsenka mukotsiliarnoi funktsii slizistoi obolochki nosovoi polosti pri khronicheskikh zabolevaniiakh legkikh.

The authors presented their first experience of radionuclide assessment of the nasal mucosal transport (NMT) in 39 patients with chronic bronchopulmonary pathology (14 adults and 25 children) to detect patients among them with suspected syndrome of primary ciliary dyskinesia (SPCD). In 18 patients the test was positive excluding SPCD as a cause of a chronic inflammatory pulmonary process. In 21 patients NMT was absent, 2 of these patients had Kartagener's triad serving as a kind of model confirming the data on NMT. The authors proposed the method as a screening test due to its non-invasiveness and simplicity in patients with chronic inflammatory pulmonary diseases in order to detect among them patients with suspected SPCD for further examination using invasive methods.

The relationships of aerosol deposition, lung size, and the rate of mucociliary clearance.

Although the effect of deposition pattern of an inhaled aerosol upon subsequent mucociliary clearance is well recognized, the influence of lung size on the rate of clearance has not been clearly defined. To examine the relationships of lung size in terms of the forced vital capacity (FVC) and aerosol deposition pattern to the rate of mucociliary clearance, we measured lung retention of an inhaled 8-micron aerosol for 120 min using a gamma camera in 13 healthy nonsmoking volunteers. The deposition pattern of the inhaled aerosol was defined in terms of the initial skew of the distribution of retained activity within the right lung (skew) and the percent retained at 24 hr (R24). Multiple regression analysis showed that FVC was responsible for approximately 36% of the variability in bronchial retention with time, with a lesser contribution from variations in deposition pattern (skew and R24). Mucociliary clearance was significantly slower (i.e., greater bronchial retention) in male subjects than in female subjects which can probably be attributed to the differences in FVC. Variations in lung sizes as well as deposition pattern should therefore be considered when making between subject comparisons of the mucociliary clearance rate.

[Radioisotopic study of the nasal muco-ciliary clearance (author's transl)]. / Etude du drainage muco-ciliaire nasal par méthode radioisotopique.

The authors propose a method for the investigation of the muco-ciliary function of the nasal mucosa: they record the motion of radioactive particles, which are put by sonic nebulisation on nasal mucosa. The average speed is calculated by means of the analysis of sequential images and by the study of the variation of activity in relation to time in several main zones. A half-clearing time of the initial activity is also measured. The average speed is 8.33 mm/minute for normal persons and 2.36 mm/minute for a group of patients. This simple, accurate, non aggressive method is used for pharmacodynamic tests.