Plasma and intracellular membrane inositol 1,4,5-trisphosphate receptors mediate the Ca2+ increase associated with the ATP-induced increase in ciliary beat frequency

An increase in intracellular free Ca2+ concentration ([Ca2+]i) has been shown to be involved in the increase in ciliary beat frequency (CBF) in response to ATP; however, the signaling pathways associated with inositol 1,4,5-trisphosphate (IP3) receptor-dependent Ca2+ mobilization remain unresolved. Using radioimmunoassay techniques, we have demonstrated the appearance of two IP3 peaks occurring 10 and 60 s after ATP addition, which was strongly correlated with a release of intracellular Ca2+ from internal stores and an influx of extracellular Ca2+, respectively. In addition, ATP-dependent Ca2+ mobilization required protein kinase C (PKC) and Ca2+/calmodulin-dependent protein kinase II activation. We found an increase in PKC activity in response to ATP, with a peak at 60 s after ATP addition. Xestospongin C, an IP3 receptor blocker, significantly diminished both the ATP-induced increase in CBF and the initial transient [Ca2+]i component. ATP addition in the presence of xestospongin C or thapsigargin revealed that the Ca2+ influx is also dependent on IP3 receptor activation. Immunofluorescence and confocal microscopic studies showed the presence of IP3 receptor types 1 and 3 in cultured ciliated cells. Immunogold electron microscopy localized IP3 receptor type 3 to the nucleus, the endoplasmic reticulum, and, interestingly, the plasma membrane. In contrast, IP3 receptor type 1 was found exclusively in the nucleus and the endoplasmic reticulum. Our study demonstrates for the first time the presence of IP3 receptor type 3 in the plasma membrane in ciliated cells and leads us to postulate that the IP3 receptor can directly trigger Ca2+ influx in response to ATP.

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Neuropeptide Y inhibits ciliary beat frequency in human ciliated cells via nPKC, independently of PKA

AJP Cell Physiology ◽

10.1152/ajpcell.1998.275.2.c440 ◽

1998 ◽

Vol 275 (2) ◽

pp. C440-C448 ◽

Cited By ~ 61

Author(s):

Lid B. Wong ◽

C. Lucy Park ◽

Donovan B. Yeates

Keyword(s):

Protein Kinase ◽

Neuropeptide Y ◽

Kinase Inhibitors ◽

Beat Frequency ◽

Specific Inhibitor ◽

Ciliary Beat Frequency ◽

Ciliated Cells ◽

Inhibitory Neurotransmitter ◽

Gf 109203X ◽

Ciliary Beat

The intracellular mechanisms whereby the inhibitory neurotransmitter neuropeptide Y (NPY) decreases ciliary beat frequency (CBF) were investigated in cultured human tracheal and bronchial ciliated cells. CBF was measured by nonstationary analysis laser light scattering. NPY at 1 and 10 μM decreased CBF from a baseline of 6.7 ± 0.5 ( n = 12) to 6.1 ± 0.5 ( P < 0.05) and 5.8 ± 0.4 ( P < 0.01) Hz, respectively. Prior application of PYX-1, an NPY antagonist, prevented the decreases of CBF induced by both doses of NPY. Two broad protein kinase C (PKC) kinase inhibitors, staurosporine and calphostin C, also abolished the NPY-induced decrease in CBF. The NPY-induced decrease in CBF was abolished by GF 109203X, a novel PKC (nPKC) isoform inhibitor, whereas this decrease in CBF was not attenuated by Gö-6976, a specific inhibitor of conventional PKC isoforms. Because pretreatment with NPY did not block the stimulation of CBF by forskolin and pretreatment with forskolin did not abolish the NPY-induced inhibition of CBF, this NPY receptor-mediated signal transduction mechanism appears to be independent of the adenylate cyclase-protein kinase A (PKA) pathway. Inhibition of Ca2+-ATPase by thapsigargin also prevented the suppression of CBF induced by subsequent application of NPY. These novel data indicate that, in cultured human epithelia, NPY decreases CBF below its basal level via the activation of an nPKC isoform and Ca2+-ATPase, independent of the activity of PKA. This is consistent with the proposition that NPY is an autonomic efferent inhibitory neurotransmitter regulating mucociliary transport.

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Effects of histamine on ciliary beat frequency of ciliated cells from guinea pigs nasal mucosa

European Archives of Oto-Rhino-Laryngology ◽

10.1007/s00405-014-3354-2 ◽

2014 ◽

Vol 272 (10) ◽

pp. 2839-2845 ◽

Cited By ~ 1

Author(s):

Fengwei An ◽

Lijun Xing ◽

Zhiqiang Zhang ◽

Lei Chen

Keyword(s):

Nasal Mucosa ◽

Guinea Pigs ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Ciliated Cells ◽

Ciliary Beat

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Control of the beat cycle of respiratory tract cilia by Ca2+ and cAMP

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1992.263.2.l232 ◽

1992 ◽

Vol 263 (2) ◽

pp. L232-L242 ◽

Cited By ~ 13

Author(s):

A. B. Lansley ◽

M. J. Sanderson ◽

E. R. Dirksen

Keyword(s):

Respiratory Tract ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Ciliated Cells ◽

Intact Cells ◽

Permeabilized Cells ◽

Whole Cells ◽

Phase Reduction ◽

Before And After ◽

Ciliary Beat

Beat frequency and the duration of the constituent recovery, effective, and rest phases of the beat cycle of respiratory tract cilia were measured photoelectronically before and after manipulation with ionomycin or isoproterenol. Both ionomycin, acting by increasing intracellular Ca2+, and isoproterenol, acting by elevating intracellular adenosine 3',5'-cyclic monophosphate (cAMP), increased beat frequency by reducing the duration of the three phases of the ciliary beat cycle in a similar manner. The addition of increasing concentrations of ATP to ciliated cells permeabilized by exposure to saponin caused a pattern of phase reduction indistinguishable from that observed in whole cells. The beat frequency of permeabilized cells was slower than that of whole cells and insensitive to changes in Ca2+ and cAMP. Ca2+ and cAMP may regulate ciliary beat frequency by acting at a common site within intact cells, possibly regulating the rate at which the axoneme can use ATP or the availability of ATP to the axoneme.

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Experience in Counting Ciliary Beat Frequency from Vital Cytologic Sampling

American Journal of Rhinology ◽

10.2500/105065889782009642 ◽

1989 ◽

Vol 3 (3) ◽

pp. 151-154

Author(s):

T. Deitmer ◽

S. Phadhana-anek

Keyword(s):

Functional State ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Time Shift ◽

Ciliated Cells ◽

Good Picture ◽

Brush Biopsy ◽

Epithelial Cell Layer ◽

Sampling Procedures ◽

Ciliary Beat

From a viable cytologic brush biopsy of the respiratory epithelium you can get an insight into the functional state of the epithelial cell layer, especially of the ciliated cells. We report on our experience of several hundred sampling procedures from the nasal and bronchial mucosa. The technique of the method is described, including the microphotometric apparatus to determine the ciliary beat frequency. We stress the importance of checking the ciliary beat frequency of the 10 most active cells of one preparation to get representative results. Futhermore it is decisive to watch them over a period of 10 seconds, considering the time shift of the ciliary beat frequency. Putting the cell solution into a counting chamber gives the possibility of differentiating viable and dead ciliated cells as well as squamous cells in the sample. These results proved to yield a good picture of the functional state of the sampled respiratory mucosal site.

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TNFαAffects Ciliary Beat Response to Increased Viscosity in Human Pediatric Airway Epithelium

BioMed Research International ◽

10.1155/2016/3628501 ◽

2016 ◽

Vol 2016 ◽

pp. 1-9 ◽

Cited By ~ 3

Author(s):

Claudia González ◽

Karla Droguett ◽

Mariana Rios ◽

Noam A. Cohen ◽

Manuel Villalón

Keyword(s):

Airway Epithelium ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Ciliated Cells ◽

Upper Airways ◽

Pediatric Airway ◽

Adenoid Tissue ◽

Before And After ◽

Autoregulatory Mechanism ◽

Ciliary Beat

In airway epithelium, mucociliary clearance (MCC) velocity depends on the ciliary beat frequency (CBF), and it is affected by mucus viscoelastic properties. Local inflammation induces secretion of cytokines (TNFα) that can alter mucus viscosity; however airway ciliated cells have an autoregulatory mechanism to prevent the collapse of CBF in response to increase in mucus viscosity, mechanism that is associated with an increment in intracellular Ca+2level (Ca2+i). We studied the effect of TNFαon the autoregulatory mechanism that regulates CBF in response to increased viscosity using dextran solutions, in ciliated cells cultured from human pediatric epithelial adenoid tissue. Cultures were treated with TNFα, before and after the viscous load was changed. TNFαtreatment produced a significantly larger decrease in CBF in cultures exposed to dextran. Furthermore, an increment inCa2+iwas observed, which was significantly larger after TNFαtreatment. In conclusion, although TNFαhas deleterious effects on ciliated cells in response to maintaining CBF after increasing viscous loading, it has a positive effect, since increasingCa2+imay prevent the MCC collapse. These findings suggest that augmented levels of TNFαassociated with an inflammatory response of the nasopharyngeal epithelium may have dual effects that contribute to maintaining the effectiveness of MCC in the upper airways.

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nNOS regulates ciliated cell polarity, ciliary beat frequency, and directional flow in mouse trachea

Life Science Alliance ◽

10.26508/lsa.202000981 ◽

2021 ◽

Vol 4 (5) ◽

pp. e202000981

Author(s):

Anatoly Mikhailik ◽

Tatyana V Michurina ◽

Krikor Dikranian ◽

Stephen Hearn ◽

Vladimir I Maxakov ◽

...

Keyword(s):

Beat Frequency ◽

Ciliated Cell ◽

Ciliary Beat Frequency ◽

Ciliary Activity ◽

Ciliated Cells ◽

No Donors ◽

Multiciliated Cells ◽

Human Disorders ◽

Low Levels ◽

Ciliary Beat

Clearance of the airway is dependent on directional mucus flow across the mucociliary epithelium, and deficient flow is implicated in a range of human disorders. Efficient flow relies on proper polarization of the multiciliated cells and sufficient ciliary beat frequency. We show that NO, produced by nNOS in the multiciliated cells of the mouse trachea, controls both the planar polarity and the ciliary beat frequency and is thereby necessary for the generation of the robust flow. The effect of nNOS on the polarity of ciliated cells relies on its interactions with the apical networks of actin and microtubules and involves RhoA activation. The action of nNOS on the beat frequency is mediated by guanylate cyclase; both NO donors and cGMP can augment fluid flow in the trachea and rescue the deficient flow in nNOS mutants. Our results link insufficient availability of NO in ciliated cells to defects in flow and ciliary activity and may thereby explain the low levels of exhaled NO in ciliopathies.

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Regulation of ciliary beat frequency by both PKA and PKG in bovine airway epithelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1998.275.4.l827 ◽

1998 ◽

Vol 275 (4) ◽

pp. L827-L835 ◽

Cited By ~ 22

Author(s):

T. A. Wyatt ◽

J. R. Spurzem ◽

K. May ◽

J. H. Sisson

Keyword(s):

Protein Kinase ◽

Epithelial Cells ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Airway Epithelial Cells ◽

Airway Epithelial ◽

Dependent Protein Kinase ◽

Dependent Protein ◽

Ciliary Beat

Ciliary beating is required for the maintenance of lung mucociliary transport. We investigated the role of cyclic nucleotide-dependent protein kinases in stimulating ciliary beat frequency (CBF) in bovine bronchial epithelial cells (BBECs). cAMP-dependent protein kinase (PKA) activity and cGMP-dependent protein kinase (PKG) activity were distinguished after DEAE-Sephacel chromatography of BBEC extracts. cAMP levels and PKA activity are increased in BBECs stimulated with 0.01–1 mM isoproterenol, with a corresponding increase in CBF. cGMP levels and PKG activity are increased in BBECs stimulated with 0.1–10 μM sodium nitroprusside, with a corresponding increase in CBF. Direct protein kinase-activating analogs of cAMP and cGMP (dibutyryl cAMP and 8-bromo-cGMP, respectively) also activate their specific kinases and stimulate CBF. Preincubation of BBECs with inhibitors of PKA or PKG [KT-5720 or Rp-8-( p-chlorophenylthio)-guanosine 3′,5′-cyclic monophosphothioate] results in the inhibition of specific kinase activity as well as in the inhibition of CBF. These studies suggest that the activation of either PKA or PKG can lead to the stimulation of CBF in bovine airway epithelium.

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LPS increase of ciliary beat frequency in respiratory ciliated cells.

The FASEB Journal ◽

10.1096/fasebj.27.1_supplement.1215.1 ◽

2013 ◽

Vol 27 (S1) ◽

Author(s):

Daniela Veronica Carreno ◽

Carmen Llados ◽

Mariana Rios ◽

Noam Cohen ◽

Manuel Villalon

Keyword(s):

Beat Frequency ◽

Ciliary Beat Frequency ◽

Ciliated Cells ◽

Ciliary Beat

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Early Mucosal Changes in Experimental Sinusitis

Otolaryngology ◽

10.1177/019459989210700405 ◽

1992 ◽

Vol 107 (4) ◽

pp. 537-548 ◽

Cited By ~ 40

Author(s):

Michael L. Hinni ◽

Thomas V. Mccaffrey ◽

Jan L. Kasperbauer

Keyword(s):

Defense Mechanism ◽

Beat Frequency ◽

Acute Infection ◽

Ciliated Cell ◽

Ciliary Beat Frequency ◽

Acute Sinusitis ◽

Ciliated Cells ◽

Predisposing Factor ◽

Sinus Mucosa ◽

Ciliary Beat

Normal mucociliary flow is a significant defense mechanism in the prevention of acute sinusitis. We have undertaken a study to examine the early sinus mucosal and mucociliary changes that occur in response to acute infection. Twenty rabbits were evaluated for 5 days after an obstructed maxillary sinus was inoculated with either Streptococcus pneumoniae, Hemophilus influenzae, Pseudomonas aeruginosa, or a sterile saline solution. Data collected included measurements of sinus mucosal ciliary beat frequency, quantitation of ciliated cell losses, and electron microscopic observations. Results demonstrate statistically significant (p < 0.05) changes in mucosal ciliary beat frequency that were either excitatory or inhibitory, depending both on the length of the infection and the specific organism. No changes in ciliary beat frequency were observed in the control animals (p > 0.55). Control animals likewise demonstrated no loss of ciliated cells from mucosal epithelium; however, dramatic losses of ciliated cells from the sinus mucosa of the experimental groups were observed. These losses occurred at different rates, depending on the infecting organism, but all infected groups demonstrated a >86% decrease in the number of viable ciliated cells from the sinus mucosa after sinusitis of 5 days duration. We conclude that a significant loss of ciliated cells from sinus mucosa and a corresponding disruption of normal mucociliary flow occurs early after exposure to pathogenic organisms and is a significant predisposing factor in the development of acute sinusitis.

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