All‐digital image capture and whole‐field analysis of ciliary beat frequency

Sisson, Joseph H.; Stoner, Julie A.; Ammons, B. A.; Wyatt, Todd A.

doi:10.1046/j.1365-2818.2003.01209.x

Cited by 261 publications

(254 citation statements)

References 13 publications

Supporting

Mentioning

249

Contrasting

Order By: Relevance

“…TRPV4 Ϫ/Ϫ ciliated tracheal cells show a reduced Ca 2ϩ signal and CBF response to changes in temperature from 24°C to 38°C, reinforcing the observation that TRPV4 activity can be coupled to CBF regulation. Interestingly, considering that maximal increase in CBF is reached within 33°C to 43°C in human and bovine airways cells (35,36), our data suggest that TRPV4 may play an important role in the control of CBF under physiological temperatures.…”

Section: Discussionmentioning

confidence: 72%

TRPV4 channel participates in receptor-operated calcium entry and ciliary beat frequency regulation in mouse airway epithelial cells

Lorenzo

Liedtke

Sanderson

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

178

171

View full text Add to dashboard Cite

The rate of mucociliary clearance in the airways is a function of ciliary beat frequency (CBF), and this, in turn, is increased by increases in intracellular calcium. The TRPV4 cation channel mediates Ca 2؉ influx in response to mechanical and osmotic stimuli in ciliated epithelia. With the use of a TRPV4-deficient mouse, we now show that TRPV4 is involved in the airways' response to physiologically relevant physical and chemical stimuli. Ciliary TRPV4 expression in tracheal epithelial cells was confirmed with immunofluorescence in TRPV4 ؉/؉ mice. Ciliated tracheal cells from TRPV4 ؊/؊ mice showed no increases in intracellular Ca 2؉ and CBF in response to the synthetic activator 4␣-phorbol 12,13-didecanoate (4␣PDD) and reduced responses to mild temperature, another TRPV4-activating stimulus. Autoregulation of CBF in response to high viscosity solutions is preserved in TRPV4 ؊/؊ despite a reduced Ca 2؉ signal. More interestingly, TRPV4 contributed to an ATPinduced increase in CBF, providing a pathway for receptor-operated Ca 2؉ entry but not store-operated Ca 2؉ entry as the former mechanism is lost in TRPV4 ؊/؊ cells. Collectively, these results suggest that TRPV4 is predominantly located in the cilia of tracheal epithelial cells and plays a key role in the transduction of physical and chemical stimuli into a Ca 2؉ signal that regulates CBF and mucociliary transport. Moreover, these studies implicate the participation of TRPV4 in receptor-operated Ca 2؉ entry.ATP ͉ trachea ͉ temperature ͉ transient receptor potential channel ͉ store-operated calcium entry I n mammalian airways, ciliated and mucus-secreting epithelial cells form a structural and functional unit that functions as a conveyor belt system for particle transport. In this analogy, cilia provide the power, whereas the mucus serves as the viscoelastic belt (1). A critical factor regulating the velocity of mucociliary transport is the ciliary beat frequency (CBF), a mechanism in which cytosolic Ca 2ϩ plays a major role (1, 2). Increases in cytosolic Ca 2ϩ are associated with increases in CBF (3, 4), although the ultimate molecular mechanism explaining CBF regulation by Ca 2ϩ remains controversial (3). Both mechanical and chemical (paracrine) stimulation of epithelial ciliated cells can lead to an increase in intracellular Ca 2ϩ concentration and the consequent enhancement of CBF (2). ATP-mediated activation of G protein-coupled receptors, typically P2Y receptors, is one of the strongest signals known to increase CBF (2). As with many other agonists of G proteincoupled receptors, ATP promotes both the release of Ca 2ϩ from inositol trisphosphate (IP 3 )-sensitive intracellular stores and Ca 2ϩ entry across the plasma membrane (4), and the latter process is more evident at micromolar concentrations of ATP where a sustained Ca 2ϩ influx occurs. The stimulation of Ca 2ϩ influx involves signaling from the depleted stores to plasma membrane Ca 2ϩ channels (store-operated calcium entry, SOCE; also known as capacitative Ca 2ϩ entry) and/or through a phospholipase...

show abstract

Section: Discussionmentioning

confidence: 72%

TRPV4 channel participates in receptor-operated calcium entry and ciliary beat frequency regulation in mouse airway epithelial cells

Lorenzo

Liedtke

Sanderson

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

178

171

View full text Add to dashboard Cite

show abstract

“…CBF was determined with Sisson-Ammons Video Analysis (39). Respiratory epithelial cells were analyzed with an Olympus IMT-2 microscope (40× phase contrast objective) equipped with a Redlake ES-310 Turbo monochrome high-speed video camera (Redlake, San Diego) set at 125 frames per second.…”

Section: Assessment Of Ciliary Function By High-speed Video Microscopymentioning

confidence: 99%

Effect of TH2 cytokines and interferon gamma on beat frequency of human respiratory cilia

et al. 2016

View full text Add to dashboard Cite

Background:In asthmatic airways secondary ciliary dyskinesia contributes to impaired mucociliary clearance. To investigate underlying mechanisms, we studied the effects of cytokines associated with asthma phenotype on the ciliary beat frequency (CBF) in a cell culture model of ciliated human respiratory epithelial cells. Methods: Nasal respiratory epithelial cells of 21 patients were used to prepare multicellular cells (spheroids) in the presence of the T helper (TH) 2 cytokines interleukin (IL)-4, IL-5, IL-9 and IL-13, and the TH1 cytokine interferon gamma (IFN-γ). CBF was determined by high-speed video microscopy. results: Addition of IL-4 and IL-13 and IL-4 + IL-13 decreased the mean CBF by 17, 21, and 22%, respectively, compared with untreated controls. Addition of IL-5 and IL-9 lead to an increase in mean CBF (20 and 10%, respectively). Lower concentrations of IFN-γ (0.1 and 1 ng/ml) decreased mean CBF and higher concentrations (10 ng/ml) increased CBF by 6%. Addition of IFN-γ to IL-13 reversed the effect of IL-13 on the CBF of spheroids. conclusion: Cytokines directly influence the ciliary function of respiratory epithelium and contribute to the impaired mucociliary clearance in asthmatic disease. Our study encourages further research to investigate IFN-γ as a treatment option in diseases with impaired mucociliary clearance like asthma.

show abstract

“…Actively beating ciliated cells from primary BBECs or tracheal rings were observed and their motion quantified using phasecontrast microscopy and computerized frequency spectrum analysis as described by Sisson et al (52). During CBF measurements, both BBECs and tracheal rings were maintained at a constant temperature (24 Ϯ 0.5°C) by a thermostatically controlled heated stage.…”

Section: Methodsmentioning

confidence: 99%

Adenosine activation of A_2Breceptor(s) is essential for stimulated epithelial ciliary motility and clearance

Allen‐Gipson¹,

Blackburn

Schneider

et al. 2011

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

sine activation of A2B receptor(s) is essential for stimulated epithelial ciliary motility and clearance. Am J Physiol Lung Cell Mol Physiol 301: L171-L180, 2011. First published May 27, 2011 doi:10.1152/ajplung.00203.2010.-Mucociliary clearance, vital to lung clearance, is dependent on cilia beat frequency (CBF), coordination of cilia, and the maintenance of periciliary fluid. Adenosine, the metabolic breakdown product of ATP, is an important modulator of ciliary motility. However, the contributions of specific adenosine receptors to key airway ciliary motility processes are unclear. We hypothesized that adenosine modulates ciliary motility via activation of its cell surface receptors (A1, A2A, A2B, or A3). To test this hypothesis, mouse tracheal rings (MTRs) excised from wild-type and adenosine receptor knockout mice (A1, A2A, A2B, or A3, respectively), and bovine ciliated bronchial epithelial cells (BBECs) were stimulated with known cilia activators, isoproterenol (ISO; 10 M) and/or procaterol (10 M), in the presence or absence of 5=-(Nethylcarboxamido) adenosine (NECA), a nonselective adenosine receptor agonist [100 nM (A1, A2A, A3); 10 M (A2B)], and CBF was measured. Cells and MTRs were also stimulated with NECA (100 nM or 10 M) in the presence and absence of adenosine deaminase inhibitor, erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (10 M). Both ISO and procaterol stimulated CBF in untreated cells and/or MTRs from both wild-type and adenosine knockout mice by ϳ3 Hz. Likewise, CBF significantly increased ϳ2-3 Hz in BBECs and wild-type MTRs stimulated with NECA. MTRs from A1, A2A, and A3 knockout mice stimulated with NECA also demonstrated an increase in CBF. However, NECA failed to stimulate CBF in MTRs from A2B knockout mice. To confirm the mechanism by which adenosine modulates CBF, protein kinase activity assays were conducted. The data revealed that NECA-stimulated CBF is mediated by the activation of cAMP-dependent PKA. Collectively, these data indicate that purinergic stimulation of CBF requires A2B adenosine receptor activation, likely via a PKA-dependent pathway. mucociliary clearance; knockout mouse model; bovine bronchial epithelial cells MUCOCILIARY TRANSPORT is an important host defense mechanism in the lung by which the coordinated beating of cilia continuously clears the lung of aspirated microorganisms and inhaled debris. This transport is dependent on the ciliary motion of airway epithelium, physicochemical properties of periciliary fluid, and mucus secretion (45, 50). Regulation of airway ciliary motility is critical because mucociliary transport is considered the first line of defense between the lung and the environment. Stressful conditions such as exercise or inflammation cause cilia in the airway to beat faster, increase clearance, and move an increased number of inhaled particles (2). As a consequence, mucociliary transport is a regulable host defense that can be activated under numerous conditions. It has been well described that ATP, a signaling molecule released by the airw...

show abstract

All‐digital image capture and whole‐field analysis of ciliary beat frequency

Cited by 261 publications

References 13 publications

TRPV4 channel participates in receptor-operated calcium entry and ciliary beat frequency regulation in mouse airway epithelial cells

TRPV4 channel participates in receptor-operated calcium entry and ciliary beat frequency regulation in mouse airway epithelial cells

Effect of TH2 cytokines and interferon gamma on beat frequency of human respiratory cilia

Adenosine activation of A_2Breceptor(s) is essential for stimulated epithelial ciliary motility and clearance

Contact Info

Product

Resources

About

All‐digital image capture and whole‐field analysis of ciliary beat frequency

Cited by 261 publications

References 13 publications

TRPV4 channel participates in receptor-operated calcium entry and ciliary beat frequency regulation in mouse airway epithelial cells

TRPV4 channel participates in receptor-operated calcium entry and ciliary beat frequency regulation in mouse airway epithelial cells

Effect of TH2 cytokines and interferon gamma on beat frequency of human respiratory cilia

Adenosine activation of A2Breceptor(s) is essential for stimulated epithelial ciliary motility and clearance

Contact Info

Product

Resources

About

Adenosine activation of A_2Breceptor(s) is essential for stimulated epithelial ciliary motility and clearance