Primary ciliary dyskinesia: evaluation using cilia beat frequency assessment via spectral analysis of digital microscopy images

Olm, Mary Anne Kowal; Kögler, João Eduardo; Macchione, Mariângela; Shoemark, Amelia; Saldiva, Paulo Hilário Nascimento; Rodrigues, Joaquim Carlos

doi:10.1152/japplphysiol.00629.2010

Cited by 31 publications

(33 citation statements)

References 43 publications

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“…The functional abnormalities observed in Agr3 2/2 mice suggest that AGR3 could be relevant for human diseases with inherited or acquired defects in ciliary beat frequency and mucociliary clearance. For example, the reductions in CBF caused by AGR3 deficiency (20% reduction in the presence of extracellular calcium and 35% after ATP stimulation) are similar to the 25 to 29% reductions in CBF that have been described for some individuals with PCD (39,40). Our findings therefore establish AGR3 as a logical candidate gene for PCD.…”

Section: Original Research Discussionsupporting

confidence: 83%

The Endoplasmic Reticulum Resident Protein AGR3. Required for Regulation of Ciliary Beat Frequency in the Airway

Bonser¹,

Schroeder²,

Ostrin

et al. 2015

Am J Respir Cell Mol Biol

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Protein disulfide isomerase (PDI) family members regulate protein folding and calcium homeostasis in the endoplasmic reticulum (ER). The PDI family member anterior gradient (AGR) 3 is expressed in the airway, but the localization, regulation, and function of AGR3 are poorly understood. Here we report that AGR3, unlike its closest homolog AGR2, is restricted to ciliated cells in the airway epithelium and is not induced by ER stress. Mice lacking AGR3 are viable and develop ciliated cells with normal-appearing cilia. However, ciliary beat frequency was lower in airways from AGR3-deficient mice compared with control mice (20% lower in the absence of stimulation and 35% lower after ATP stimulation). AGR3 deficiency had no detectable effects on ciliary beat frequency (CBF) when airways were perfused with a calcium-free solution, suggesting that AGR3 is required for calcium-mediated regulation of ciliary function. Decreased CBF was associated with impaired mucociliary clearance in AGR3-deficient airways. We conclude that AGR3 is a specialized member of the PDI family that plays an unexpected role in the regulation of CBF and mucociliary clearance in the airway.

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Section: Original Research Discussionsupporting

confidence: 83%

The Endoplasmic Reticulum Resident Protein AGR3. Required for Regulation of Ciliary Beat Frequency in the Airway

Bonser¹,

Schroeder²,

Ostrin

et al. 2015

Am J Respir Cell Mol Biol

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“…The presence of harmonics in the spectrum had already been pointed out [12] but was never used before in order to directly improve the CBF estimation. Furthermore, positioning the sub-image over the region of interest, similar to the line of interest (LOI) [9,13], is still an open question. In the literature, this positioning remains manual as the subsequent processing, and finally subjective.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, the second evaluation will provide an analysis of sub-image location and size influences. In both cases, the range of the exhaustive search for the optimal frequency will be [6][7][8][9][10][11][12][13][14][15][16][17][18][19][20] Hz with a resolution of 0.01 Hz, using a zero padding technique for the FFT, and for the three methods.…”

Section: Evaluation Of Methodsmentioning

confidence: 99%

“…In this respect, standard signal processing tools as Fourier analysis [21] may be helpful. Recent studies have proposed methods based on visual inspection [11], classical Fourier analysis [11,13] and pixel average [4,11,19]. However, for long experiments, frequencies computed from visual measurement have to be used carefully because of potential bias.…”

Section: Introductionmentioning

confidence: 99%

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Robust estimation of the motile cilia beating frequency

Meste

Brau

Guyon

2015

Med Biol Eng Comput

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The estimation of the cilia beating frequency (CBF) is of great interest in understanding how the CBF modulates liquid fluxes and how it is controlled by the ciliated cell intra- and/or extracellular medium composition in physiological processes. Motion artifacts and camera defaults may hinder the computation of the frequency variations during long-lasting experiments. We have developed a new analysis approach consisting of a preliminary corrective step (removal of a grid pattern on the image sequence and shift compensation), followed by a harmonic model of the observed cilia using a maximum likelihood estimator framework. It is shown that a more accurate estimation of the frequency can be obtained by averaging the squared Fourier transform of individual pixels followed by a particular summation over the different frequencies, namely the compressed spectrum. Robustness of the proposed method over traditional approaches is shown by several examples and simulations. The method is then applied to images of samples containing ciliated ependymal cells located in the third cerebral ventricle of mouse brains, showing that even small variations in CBF in response to changes in the amount of oxygenation, pH or glucose were clearly visible in the computed frequencies. As a conclusion, this method reveals a fine metabolic tuning of the cilia beating in ependimocytes lining the third cerebral ventricle. Such regulations are likely to participate in homeostatic mechanisms regulating CSF movements and brain energy supply.

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“…It is recognized that standard light microscopic analysis is not sufficient (66). Even with highspeed video microscopy, there are recognized limitations due to: (1) effects of infection and inflammation on cilia; (2) disruption of epithelium during sample collection; (3) lack of quantitative data to allow rigorous interpretation of waveform analyses; and (4) overlap of ciliary beat frequency between PCD and diseasecontrol and normal subjects (67,68). Approaches to improve accuracy include ciliated air-liquid cultures to reduce the effects of infection/inflammation, and cooling the specimen to slow cilia beat frequency for waveform analysis; however, these modifications do not eliminate all of the limitations (69-71).…”

Section: Ciliary Beat Frequency and Motilitymentioning

confidence: 99%

Primary Ciliary Dyskinesia. Recent Advances in Diagnostics, Genetics, and Characterization of Clinical Disease

Knowles¹,

Daniels²,

Davis

et al. 2013

Am J Respir Crit Care Med

435

460

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Primary ciliary dyskinesia (PCD) is a genetically heterogeneous recessive disorder of motile cilia that leads to oto-sino-pulmonary diseases and organ laterality defects in approximately 50% of cases. The estimated incidence of PCD is approximately 1 per 15,000 births, but the prevalence of PCD is difficult to determine, primarily because of limitations in diagnostic methods that focus on testing ciliary ultrastructure and function. Diagnostic capabilities have recently benefitted from (1) documentation of low nasal nitric oxide production in PCD and (2) discovery of biallelic mutations in multiple PCD-causing genes. The use of these complementary diagnostic approaches shows that at least 30% of patients with PCD have normal ciliary ultrastructure. More accurate identification of patients with PCD has also allowed definition of a strong clinical phenotype, which includes neonatal respiratory distress in .80% of cases, daily nasal congestion and wet cough starting soon after birth, and early development of recurrent/chronic middle-ear and sinus disease. Recent studies, using advanced imaging and pulmonary physiologic assessments, clearly demonstrate early onset of lung disease in PCD, with abnormal air flow mechanics by age 6-8 years that is similar to cystic fibrosis, and age-dependent onset of bronchiectasis. The treatment of PCD is not standardized, and there are no validated PCD-specific therapies. Most patients with PCD receive suboptimal management, which should include airway clearance, regular surveillance of pulmonary function and respiratory microbiology, and use of antibiotics targeted to pathogens. The PCD Foundation is developing a network of clinical centers, which should improve diagnosis and management of PCD.

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Primary ciliary dyskinesia: evaluation using cilia beat frequency assessment via spectral analysis of digital microscopy images

Cited by 31 publications

References 43 publications

The Endoplasmic Reticulum Resident Protein AGR3. Required for Regulation of Ciliary Beat Frequency in the Airway

The Endoplasmic Reticulum Resident Protein AGR3. Required for Regulation of Ciliary Beat Frequency in the Airway

Robust estimation of the motile cilia beating frequency

Primary Ciliary Dyskinesia. Recent Advances in Diagnostics, Genetics, and Characterization of Clinical Disease

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