Functional cross talk after activation of P2 and P1 receptors in oviductal ciliated cells

2000 ◽  
Vol 279 (3) ◽  
pp. C658-C669 ◽  
Author(s):  
Bernardo Morales ◽  
Nelson Barrera ◽  
Pablo Uribe ◽  
Claudio Mora ◽  
Manuel Villalón
Keyword(s):  
Cross Talk ◽  
Adenosine Receptors ◽  
Beat Frequency ◽  
Ciliated Cell ◽  
Ciliary Beat Frequency ◽  
Ciliary Activity ◽  
Dependent Manner ◽  
Ciliated Cells ◽  
P1 Receptors ◽  
Intracellular Ca

The presence of ATP and adenosine receptors and their role in controlling ciliary activity in oviductal ciliated cells was studied by measuring the ciliary beat frequency (CBF) in oviductal tissue cultures. ATP, adenosine, and related compounds increased the CBF in a dose-dependent manner. We established that P2 receptors of subtype 2Y2 and P1 receptors of subtype A2a mediated the responses to ATP and adenosine, respectively. We found evidence to suggest that stimulation of ciliary activity by ATP requires d- myo-inositol 1,4,5-trisphosphate [Ins(1,4,5) P 3] metabolism, intracellular Ca2+ mobilization, and protein kinase C activation. On the other hand, the adenosine effect is mediated by activation of a Gs protein-dependent pathway that enhances cAMP intracellular levels. To study the interaction between P2 and P1 receptors, cells were stimulated simultaneously with both agonists. We observed a synergistic increase of the CBF even at agonist concentrations (100 nM) that did not produce a significant response when added separately to the culture. Furthermore, a blocker of the cAMP pathway produced a reduction of the ATP response, whereas a blocker of the Ins(1,4,5) P 3 pathway also produced an inhibition of the adenosine response. Our evidence demonstrates that both ATP and adenosine receptors are present in a single ciliated cell and that a mechanism of cross talk could operate in the transduction pathways to control ciliary activity.

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

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.

scholarly journals PKA induces Ca2+ release and enhances ciliary beat frequency in a Ca2+-dependent and -independent manner

1998 ◽  
Vol 275 (3) ◽  
pp. C790-C797 ◽  
Author(s):  
Alex Braiman ◽  
Orna Zagoory ◽  
Zvi Priel
Keyword(s):  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Ciliary Activity ◽  
Experimental Conditions ◽  
Independent Manner ◽  
Ciliary Beating ◽  
Independent Mechanism ◽  
Intracellular Ca ◽  
Ciliary Beat

The intent of this work was to evaluate the role of cAMP in regulation of ciliary activity in frog mucociliary epithelium and to examine the possibility of cross talk between the cAMP- and Ca2+-dependent pathways in that regulation. Forskolin and dibutyryl cAMP induced strong transient intracellular Ca2+ concentration ([Ca2+]i) elevation and strong ciliary beat frequency enhancement with prolonged stabilization at an elevated plateau. The response was not affected by reduction of extracellular Ca2+concentration. The elevation in [Ca2+]iwas canceled by pretreatment with 1,2-bis(2-aminophenoxy)ethane- N, N, N′, N′-tetraacetic acid-AM, thapsigargin, and a phospholipase C inhibitor, U-73122. Under those experimental conditions, forskolin raised the beat frequency to a moderately elevated plateau, whereas the initial strong rise in frequency was completely abolished. All effects were canceled by H-89, a selective protein kinase A (PKA) inhibitor. The results suggest a dual role for PKA in ciliary regulation. PKA releases Ca2+ from intracellular stores, strongly activating ciliary beating, and, concurrently, produces moderate prolonged enhancement of the beat frequency by a Ca2+-independent mechanism.

Nitric oxide activation by progesterone suppresses ATP-induced ciliary activity in oviductal ciliated cells

2018 ◽  
Vol 30 (12) ◽  
pp. 1666
Author(s):  
Bredford Kerr ◽  
Mariana Ríos ◽  
Karla Droguett ◽  
Manuel Villalón
Keyword(s):  
Nitric Oxide ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Ciliary Activity ◽  
No Production ◽  
Ciliated Cells ◽  
Reproductive Process ◽  
Cell Functions ◽  
Intracellular Messengers ◽  
Local Release

Ciliary beat frequency (CBF) regulates the oviductal transport of oocytes and embryos, which are important components of the reproductive process. Local release of ATP transiently increases CBF by increasing [Ca2+]i. Ovarian hormones also regulate ciliary activity and oviductal transport. Progesterone (P4) induces nitric oxide (NO) production and high P4 concentrations induce ciliary dysfunction. However, the mechanism by which P4 affects CBF has not been elucidated. To evaluate the role of P4 in NO production and its effect on ATP-induced increases in CBF, we measured CBF, NO concentrations and [Ca2+]i in cultures of oviductal ciliated cells treated with P4 or NO signalling-related molecules. ATP induced a [Ca2+]i peak, followed by an increase in NO concentrations that were temporally correlated with the decreased phase of the transiently increased CBF. Furthermore, P4 increased the expression of nitric oxide synthases (iNOS and nNOS) and reduced the ATP-induced increase in CBF via a mechanism that involves the NO signalling pathway. These results have improved our knowledge about intracellular messengers controlling CBF and showed that NO attenuates oviduct cell functions. Furthermore, we showed that P4 regulates neurotransmitter (ATP) actions on CBF via the NO pathway, which could explain pathologies where oviductal transport is altered and fertility decreased.

Effect of Substance P on Ciliary Beat Frequency in Human Adenoid Explants

1992 ◽  
Vol 107 (4) ◽  
pp. 553-557 ◽  
Author(s):  
Paul A. Staskowski ◽  
Thomas V. Mccaffrey
Keyword(s):  
Substance P ◽  
Culture Media ◽  
Beat Frequency ◽  
Upper Airway ◽  
Ciliary Beat Frequency ◽  
Fast Fourier Transformation ◽  
Protective Mechanism ◽  
Ciliary Activity ◽  
Dependent Manner ◽  
Ciliary Beat

This study investigated the direct effects of substance P (SP) on ciliary beat frequency of human upper airway mucosa. Human adenoid explant tissue was maintained in serum free culture media, MCDB153. Ciliated epithelial cells were observed with phase-contrast microscopy and ciliary activity was measured using a photometric technique. Oscillations in transmitted light caused by ciliary beating were recorded and modal ciliary beat frequency was determined by fast Fourier transformation. Specimens were treated with SP at concentrations of 10−4, 10−5, 10−6, and 10−7 mol/L and with equal molar solutions of SP and (D-Pro2, D-Trp7,9)-SP, a SP antagonist. Substance P was found to increase ciliary beat frequency in a dose-dependent manner with a maximum increase of 12.1%. This effect was not seen with solutions containing (D-Pro2, D-Trp7,9)-SP. This suggests that SP exerts a direct stimulatory effect on ciliated cells of the upper airway. Because SP is known to be released in the upper airway in response to chemical irritation, it is presumed that the stimulatory effect of SP on mucosal cells provides a protective mechanism against inhaled irritants.

Early Mucosal Changes in Experimental Sinusitis

1992 ◽  
Vol 107 (4) ◽  
pp. 537-548 ◽  
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.

scholarly journals TRPV4 channel is involved in the coupling of fluid viscosity changes to epithelial ciliary activity

2005 ◽  
Vol 168 (6) ◽  
pp. 869-874 ◽  
Author(s):  
Yaniré N. Andrade ◽  
Jacqueline Fernandes ◽  
Esther Vázquez ◽  
José M. Fernández-Fernández ◽  
Maite Arniges ◽  
...  
Keyword(s):  
Transient Receptor Potential ◽  
Beat Frequency ◽  
Receptor Potential ◽  
Ciliary Beat Frequency ◽  
Transient Receptor Potential Vanilloid ◽  
Ciliary Activity ◽  
Fluid Viscosity ◽  
Ciliated Cells ◽  
Trpv4 Channel ◽  
Viscosity Changes

Autoregulation of the ciliary beat frequency (CBF) has been proposed as the mechanism used by epithelial ciliated cells to maintain the CBF and prevent the collapse of mucociliary transport under conditions of varying mucus viscosity. Despite the relevance of this regulatory response to the pathophysiology of airways and reproductive tract, the underlying cellular and molecular aspects remain unknown. Hamster oviductal ciliated cells express the transient receptor potential vanilloid 4 (TRPV4) channel, which is activated by increased viscous load involving a phospholipase A2–dependent pathway. TRPV4-transfected HeLa cells also increased their cationic currents in response to high viscous load. This mechanical activation is prevented in native ciliated cells loaded with a TRPV4 antibody. Application of the TRPV4 synthetic ligand 4α-phorbol 12,13-didecanoate increased cationic currents, intracellular Ca2+, and the CBF in the absence of a viscous load. Therefore, TRPV4 emerges as a candidate to participate in the coupling of fluid viscosity changes to the generation of the Ca2+ signal required for the autoregulation of CBF.

scholarly journals Stimulation of cilia beat frequency by serotonin is mediated by a Ca2+ influx in ciliated cells of Helisoma trivolvis embryos

1996 ◽  
Vol 199 (5) ◽  
pp. 1105-1113 ◽  
Author(s):  
K Christopher ◽  
J Chang ◽  
J Goldberg
Keyword(s):  
Cyclic Amp ◽  
Serotonin Receptor ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Time Lapse ◽  
Channel Blockers ◽  
Dependent Manner ◽  
Ciliated Cells ◽  
Helisoma Trivolvis ◽  
Transduction Mechanisms

Serotonin (5-HT) has been established as a regulator of ciliary beating in numerous systems. In early embryos of Helisoma trivolvis, a cilia-driven rotational movement is modulated by the release of endogenous serotonin from a pair of embryonic neurons, ENC1s, directly onto the ciliated epithelium. The present study was undertaken to examine the signal transduction mechanisms underlying serotonin-mediated cilio-excitation in Helisoma trivolvis embryos. Using time-lapse videomicroscopy, the ciliary beat frequency (CBF) of cultured embryonic ciliated cells was measured in response to various pharmacological manipulations. Serotonin increased CBF in a dose-dependent manner. Addition of 8-bromo-cyclic AMP, isobutylmethylxanthine (IBMX) or a combination of forskolin and IBMX, treatments that elevate the concentration of intracellular cyclic AMP, did not mimic the serotonin-induced increase in CBF. Thus, cyclic AMP does not appear to be involved in the regulation of CBF in this system. In contrast, depolarizing the cells with KCl or veratridine, and artificially raising the intracellular Ca2+ concentration with thapsigargin or A23187, caused a serotonin-like increase in CBF. Furthermore, the serotonin response was abolished in a Ca2+-depleted medium or in a medium containing the L-type Ca2+ channel blockers verapamil or nifedipine. These results suggest that serotonin-stimulated cilio-excitation in cultured Helisoma trivolvis cells involves an influx of Ca2+ to increase intracellular Ca2+ concentration. The link between serotonin-receptor binding and Ca2+ influx in these cells has yet to be determined.

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

2014 ◽  
Vol 272 (10) ◽  
pp. 2839-2845 ◽  
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

Polyglutamylation and polyglycylation of alpha- and beta-tubulins during in vitro ciliated cell differentiation of human respiratory epithelial cells

1999 ◽  
Vol 112 (23) ◽  
pp. 4357-4366 ◽  
Author(s):  
K. Million ◽  
J. Larcher ◽  
J. Laoukili ◽  
D. Bourguignon ◽  
F. Marano ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Differentiation ◽  
Epithelial Cells ◽  
Beat Frequency ◽  
Ciliated Cell ◽  
Early Event ◽  
Functional Assay ◽  
Ciliated Cells ◽  
Human Nasal Epithelial Cells ◽  
Centriole Assembly

Tubulins are the major proteins within centriolar and axonemal structures. In all cell types studied so far, numerous alpha- and beta-tubulin isoforms are generated both by expression of a multigenic family and various post-translational modifications. We have developed a primary culture of human nasal epithelial cells where the ciliated cell differentiation process has been observed and quantified. We have used this system to study several properties concerning polyglutamylation and polyglycylation of tubulin. GT335, a monoclonal antibody directed against glutamylated tubulins, stained the centriole/basal bodies and the axonemes of ciliated cells, and the centrioles of non-ciliated cells. By contrast, axonemal but not centriolar tubulins were polyglycylated. Several polyglutamylated and polyglycylated tubulin isotypes were detected by two-dimensional electrophoresis, using GT335 and a specific monoclonal antibody (TAP952) directed against short polyglycyl chains. Immunoelectron microscopy experiments revealed that polyglycylation only affected axonemal tubulin. Using the same technical approach, polyglutamylation was shown to be an early event in the centriole assembly process, as gold particles were detected in fibrogranular material corresponding to the first cytoplasmic structures involved in centriologenesis. In a functional assay, GT335 and TAP952 had a dose-dependent inhibitory effect on ciliary beat frequency. TAP952 had only a weak effect while GT335 treatment led to a total arrest of beating. These results strongly suggest that in human ciliated epithelial cells, tubulin polyglycylation has only a structural role in cilia axonemes, while polyglutamylation may have a function both in centriole assembly and in cilia activity.

Differential effects of UTP, ATP, and adenosine on ciliary activity of human nasal epithelial cells

2001 ◽  
Vol 280 (6) ◽  
pp. C1485-C1497 ◽  
Author(s):  
Diane M. Morse ◽  
Jennifer L. Smullen ◽  
C. William Davis
Keyword(s):  
Epithelial Cells ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Ciliary Activity ◽  
Maximal Effect ◽  
Ciliary Beating ◽  
Mediated Effects ◽  
Human Nasal Epithelium ◽  
Human Nasal Epithelial Cells ◽  
Sustained Responses

The purinergic regulation of ciliary activity was studied using small, continuously superfused explants of human nasal epithelium. The P2Y2 purinoceptor (P2Y2-R) was identified as the major purinoceptor regulating ciliary beat frequency (CBF); UTP (EC50 = 4.7 μM), ATP, and adenosine-5′- O-(3-thiotriphosphate) elicited similar maximal responses, approximately twofold over baseline. ATP, however, elicited a post-peak sustained plateau in CBF (1.83 ± 0.1-fold), whereas the post-peak CBF response to UTP declined over 15 min to a low-level plateau (1.36 ± 0.16-fold). UDP also stimulated ciliary beating, probably via P2Y6-R, with a maximal effect approximately one-half that elicited by P2Y2-R stimulation. Not indicated were P2Y1-R-, P2Y4-R-, or P2Y11-R-mediated effects. A2B-receptor agonists elicited sustained responses in CBF approximately equal to those from UTP/ATP [5′-( N-ethylcarboxamido)adenosine, EC50 = 0.09 μM; adenosine, EC50 = 0.7 μM]. Surprisingly, ADP elicited a sustained stimulation in CBF. The ADP effect and the post-peak sustained portion of the ATP response in CBF were inhibited by the A2-R antagonist 8-( p-sulfophenyl)theophylline. Hence, ATP affects ciliary activity through P2Y2-R and, after an apparent ectohydrolysis to adenosine, through A2BAR.

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