Biomedical subjects

Z Priel

Publications and source records attributed to Z Priel.

At least 19 recordsLinked to original sources

PubMed1998

Extracellular ATP directly gates a cation-selective channel in rabbit airway ciliated epithelial cells.

1. A membrane conductance activated by extracellular ATP was identified and characterized in freshly dissociated rabbit airway ciliated cells using the whole-cell and outside-out patch configurations of the patch-clamp technique. 2. In solutions designed to maximize currents through voltage-gated calcium channels, there were no indications of voltage-gated Ba2+ currents. 3. Extracellular ATP (but not UTP or ADP) activated a membrane conductance which remained activated for several minutes in the presence of ATP. The conductance was permeable to monovalent and divalent cations with approximate relative permeabilities (P) for PBa : PCs : PTEA of 4 : 1 : 0.1. Permeability to Cl- was negligible. 4. Including GDP-beta-S in the intracellular solution did not inhibit the effects of ATP, nor did GTP-gamma-S irreversibly activate the conductance. 5. In outside-out membrane patches, with GDP-beta-S in the pipette solution, ATP activated ion channels which had a chord conductance of approximately 6 pS in symmetrical 150 mM CsCl solutions at -120 mV. 6. Suramin (100 microM) inhibited the whole-cell currents activated by ATP (200 microM) by 93 +/- 3 %. Similar effects of suramin were observed on ATP-activated channels in outside-out membrane patches. 7. Extracellular ATP had a priming action on the response to subsequent exposure to ATP. At -40 mV, the time to half-maximal current activation (t1/2) was 46 +/- 9 s during the first exposure to 200 microM ATP and decreased to 5 +/- 3 s during a second exposure to the same concentration of ATP. The priming action of ATP was not inhibited by including GDP-beta-S in the intracellular solution. 8. The initial rate of activation increased with the concentration of ATP, and was voltage sensitive. During the first exposure to 200 microM ATP, t1/2 at +40 mV was 4-fold longer than t1/2 at -40 mV. 9. Half-maximal activation of the conductance shifted from 210 +/- 30 to 14 +/- 4 microM added ATP when CaCl2 in the extracellular solution was reduced from 1.58 to 0. 01 mM. The Hill coefficient for ATP was 1.2 in both solutions.10. These observations suggest that a form of ATP uncomplexed with divalent cations directly gates an ion channel (P2X receptor) in rabbit airway ciliated cells, which serves as a pathway for Ca2+ influx. This purinoceptor may contribute to sustained ciliary activation during prolonged exposures to ATP.

Adenosine Diphosphate↗

PubMed1998

Effect of viscosity on metachrony in mucus propelling cilia.

In the present work we report that increasing the viscosity of the medium caused not only a decrease in the ciliary beat frequency but also changes in the metachrony and correlation between cilia. The study was performed using double and triple simultaneous photoelectric measurements on cultured ciliary cells from the frog esophagus in the viscosity range of 1-2,000 cp. We observed that increasing the viscosity intensified the fluctuations in all the measured parameters. Ciliary beat frequency decreased moderately. Even at quite high viscosities (circa 2000 cp.), cilia were still active with beating frequencies of 3-5 Hz. In addition, the degree of correlation between cilia parallel to the effective stroke direction (ESD) decreased, while that perpendicular to the ESD at a low range of viscosities remained unchanged and even increased at high viscosities. Medium viscosities in the range of 30-1,500 cp. altered the metachronal wave properties of cultured frog esophagus. The metachronal wavelength increased by up to 50%, and the wave direction changed towards more orthoplectic type of coordination. According to our recently suggested model [Gheber and Priel, 1990: Cell Motil. Cytoskeleton 16:167-181], these effects can be explained by a decrease in the temporal asymmetry of the ciliary beat. Since similar results were observed in water propelling cilia of Paramecium subjected to medium viscosity ranges of up to 40 cp. [Machemer, 1972: J. Exp. Biol. 57:239-259], we conclude that hydrodynamic interactions govern the metachronal wave properties of both mucus and water propelling cilia, though mucus propelling cilia, with their better adaptation to increased load, are affected at much higher viscosities than water propelling cilia.

Animals↗

PubMed1998

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

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+]i was 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.

Animals↗

PubMed1997

A realistic model of biphasic calcium transients in electrically nonexcitable cells.

In many electrically nonexcitable cells, the release of calcium from internal stores is followed by a much slower phase in which the intracellular calcium concentration decreases gradually to a sustained value higher than the concentration before stimulation. This elevated calcium plateau has been shown to be the result of calcium influx. The model presented in this work describes a system consisting of a cytoplasmic calcium store and a plasma membrane calcium channel, both excitable by a membrane receptor; a fast cytoplasmic calcium buffer; and calcium pumps in both the calcium store and cellular membranes. Inherent difficulties in the numerical evaluation of the model, caused by very large calcium fluxes across the store membrane, were overcome by analytically separating the fast processes of calcium release from the slower processes of calcium cycling across the plasma membrane. This enabled the simulation of realistic biphasic calcium transients similar to those observed experimentally. The model predicted 1) a strong correlation between the rate of calcium cycling across the plasma membrane and the rate of calcium decay; and 2) a dependence on the level of cell excitation of the maximum rise in cytoplasmic calcium concentration, the level of the elevated calcium plateau, and the rate of calcium decay. Using the model, we simulated the washout of agonist from the bathing solution and the depletion of the calcium store by a pharmacological agent (such as thapsigargin) under several experimental conditions.

Animals↗

PubMed1997

Extraction of cilium beat parameters by the combined application of photoelectric measurements and computer simulation.

Photoelectric signals were created and used to investigate the features of the signals as a function of the ciliary beat parameters. Moreover, correlation between the simulated and the measured signals permitted measurement of the cilium beat parameters. The simulations of the signals were based on generation of a series of time-frozen top-view frames of an active ciliary area and determination of the amount of light passing through an observation area in each of these frames. All the factors that might contribute to the shape of the signals, namely, partial ciliary transmittance of light, three-dimensional ciliary beat (composed of recovery, effective, and pause parts), phase distribution on the ciliary surface, and the large number of cilia that contribute to the photoelectric signal, were taken into account in generation of the signals. Changes in the ciliary parameters influenced the shape of the photoelectric signals, and the different phases of the beat could not be directly and unequivocally identified in the signals. The degree of temporal asymmetry of the beat and the portion of the cycle occupied by the pause significantly influenced the shapes of both the lower and the upper parts of the signal and the slopes of the signal. Increases in the angle of the arc swept by the cilium during the effective stroke smoothed the signals and increased the duration of the upper part of the signal. The angle of the arc projected by the cilium onto the cell surface during the recovery stroke had minor effects on the signal's shape. Characteristics of the metachronal wave also influenced the signal's shape markedly. Decreases in ciliary spacing smoothed the signals, whereas ciliary length had a minor influence on the simulated photoelectric signals. Comparison of the simulated and the measured signals showed that the beat parameters of the best-fitting simulated signals converged to values that agree well with the accepted range of beat parameters in mucociliary systems.

Animals↗

PubMed1997

Protein kinase C induced calcium influx and sustained enhancement of ciliary beating by extracellular ATP.

The major purpose of this work was to determine protein kinase C (PKC) influence on ciliary beat frequency (CBF) and to assess participation of PKC in purinergic ciliary stimulation. The experiments were performed by simultaneous measurement of [Ca2+]i and CBF on tissue culture of frog esophagus epithelium. The PKC activators TPA and DiC8 produced significant elevation of [Ca2+]i and strong frequency enhancement. The calcium elevation was inhibited by lowering the extracellular calcium level, or by La3+, but was unaffected by verapamil and the phospholipase C inhibitor U-73122, suggesting that Ca2+ influx was via non-voltage-operated calcium channels. The inhibition of [Ca2+]i elevation resulted in corresponding inhibition of CBF enhancement. The effect of TPA was blocked by the selective PKC inhibitors chelerythrine, calphostin C, and GF109203X, and by the enzyme downregulation. The downregulation of PKC, or the enzyme inhibitors did not affect the immediate response to extracellular ATP but caused rapid decay of initially stimulated [Ca2+]i and CBF to the basal level. These results suggest that PKC produces CBF enhancement via activation of calcium influx through non-voltage-operated calcium channels. This calcium influx seems to be responsible for the duration of ciliary stimulation produced by the extracellular ATP.

Adenosine Triphosphate↗

PubMed1997

Na(+)-K(+)-ATPase in frog esophagus mucociliary cell membranes: inhibition by protein kinase C activation.

We examined protein kinase C (PKC)-dependent regulation of Na(+)-K(+)-ATPase in frog mucociliary cells. Activation of PKC by 12-O-tetradecanoylphorbol-13-acetate (TPA) or 1,2-dioctanoyl-sn-glycerol (diC8) either in intact cells or isolated membranes resulted in a specific inhibition of Na(+)-K(+)-ATPase activity by approximately 25-45%. The inhibitory effects in membranes exhibited time dependence and dose dependence [half-maximal inhibition concentration (IC50) = 0.5 +/- 0.1 nM and 2.4 +/- 0.2 microM, respectively, for TPA and diC8] and were not influenced by Ca2+. Analysis of the ouabain inhibition pattern revealed the presence of two Na(+)-K(+)-ATPase isoforms with IC50 values for cardiac glycoside of 2.6 +/- 0.8 nM and 409 +/- 65 nM, respectively. Most importantly, the isoform possessing a higher affinity for ouabain was almost completely inhibited by TPA, whereas its counterpart was hardly sensitive to the PKC activator. The results suggest that, in frog mucociliary cells, PKC regulates Na(+)-K(+)-ATPase and that this action is related to the specific Na(+)-K(+)-ATPase isoform.

4-Nitrophenylphosphatase↗

PubMed1996

Purinergic stimulation of rabbit ciliated airway epithelia: control by multiple calcium sources.

1. Simultaneous measurements of average intracellular calcium concentration ([Ca2+]i) and ciliary beat frequency (CBF) were carried out on ciliated rabbit tracheal cells in order to determine quantitatively the role of calcium in the regulation of mucus-transporting cilia. 2. Extracellular ATP caused a rapid increase in both [Ca2+]i and CBF in the 0.1-1000 microM concentration range. The rise in [Ca2+]i levelled off to an elevated [Ca2+]i plateau while the cilia remained in a high activation state. The magnitude of the rise in [Ca2+]i and CBF as well as the value of the elevated [Ca2+]i plateau and the value of the sustained CBF were dependent on the concentration of ATP in the solution. 3. No correlation was found between the mean values of [Ca2+]i and CBF at rest but a sigmoidal relationship was found to exist between the maximal rises of these parameters following excitation with extracellular ATP. This sigmoidal correlation incorporated the experiments where [Ca2+]i rise was induced by depletion of internal calcium stores with thapsigargin or by entry of calcium induced by ionomycin. 4. Extracellular ATP caused both the release of calcium from internal stores and calcium influx from the extracellular solution. The release of calcium was identified as originating from a thapsigargin-sensitive and a thapsigargin-insensitive calcium store. It is suggested that the release of calcium from these stores induces the initial rise in CBF. 5. The sustained activation of the cilia and elevated calcium plateau were found to be the result of the extracellular ATP-induced calcium influx. This calcium influx was insensitive to the voltage-gated calcium channel inhibitors verapamil and diltiazem, but was completely eliminated by lowering the extracellular calcium concentration to 0.1 microM. 6. We propose that the initial jump in the CBF is mediated by the calcium released from a thapsigargin-insensitive calcium store adjacent to the cilia, while the later, and longer, rise in CBF is the result of the calcium emanating from the thapsigargin-sensitive store which is positioned further away from the cilia within the cell cytoplasm. The calcium influx that follows is responsible for sustaining the cilia at a high level of excitation.

Adenosine Triphosphate↗

PubMed1996

Purinergically induced membrane fluidization in ciliary cells: characterization and control by calcium and membrane potential.

To examine the role of membrane dynamics in transmembrane signal transduction, we studied changes in membrane fluidity in mucociliary tissues from frog palate and esophagus epithelia stimulated by extracellular ATP. Micromolar concentrations of ATP induced strong changes in fluorescence polarization, possibly indicating membrane fluidization. This effect was dosage dependent, reaching a maximum at 10-microM ATP. It was dependent on the presence of extracellular Ca2+ (or Mg2+), though it was insensitive to inhibitors of voltage-gated calcium channels. It was inhibited by thapsigargin and by ionomycin (at low extracellular Ca2+ concentration), both of which deplete Ca2+ stores. It was inhibited by the calcium-activated potassium channel inhibitors quinidine, charybdotoxin, and apamine and was reduced considerably by replacement of extracellular Na+ with K+. Hyperpolarization, or depolarization, of the mucociliary membrane induced membrane fluidization. The degree of membrane fluidization depended on the degree of hyperpolarization or depolarization of the ciliary membrane potential and was considerably lower than the effect induced by extracellular ATP. These results indicate that appreciable membrane fluidization induced by extracellular ATP depends both on an increase in intracellular Ca2+, mainly from its internal stores, and on hyperpolarization of the membrane. Calcium-dependent potassium channels couple the two effects. In light of recent results on the enhancement of ciliary beat frequency, it would appear that extracellular ATP-induced changes both in ciliary beat frequency and in membrane fluidity are triggered by similar signal transduction pathways.

Adenosine Triphosphate↗

PubMed1995

Extracellular ATP binding proteins as potential receptors in mucociliary epithelium: characterization using [32P]3'-O-(4-benzoyl)benzoyl ATP, a photoaffinity label.

3'-O-(4-benzoyl)benzoyl ATP (BzATP) was used as a photoaffinity analog of ATP to label potential ATP receptors in ciliated cells. Like ATP, without photoactivation, BzATP stimulated the ciliary beat frequency in tissue culture up to threefold. Irradiation of intact cells in the presence of [alpha-32P]BzATP followed by SDS-PAGE and autoradiography revealed two labeled proteins with molecular masses of 46 and 96 kDa (p46 and p96). Photolabeling of both proteins was susceptible to digestion with trypsin, implying that the labeled proteins are at least partially exposed on the extracellular surface of the plasma membrane. The dependence of 32P incorporation in both proteins on [alpha-32P]BzATP concentration was similar. Labeling of p46 but not p96 required Ca2+ or Mg2+. Various nucleotides stimulated the ciliary frequency, and inhibited the photolabeling of p46 and p96. The rank order of apparent affinity for p46 is: ATP approximately equal to ADP > GTP gamma S > ADP beta S, UTP, 2MeSATP, AMP-PNP > AMP-PCP > AMP > adenosine; for p96 it is: ADP approximately equal to ADP beta S > or = ATP >> AMP-PCP, AMP-PNP > GTP gamma S > or = AMP > 2MeSATP, UTP, adenosine. The rank of stimulation of ciliary beat frequency is: ADP beta S, UTP > or = 2MeSATP, GTP gamma S, AMP-PNP, ATP > or = ADP > AMP-PCP > adenosine > AMP. These results suggest the involvement of p46 in the stimulatory effect of extracellular ATP on the ciliary beat, as a P2 purinoceptor. On the other hand, p96 may represent a P2 purinoceptor or an ectonucleotidase.

Adenosine Triphosphate↗

PubMed1995

Extracellular ATP induces hyperpolarization and motility stimulation of ciliary cells.

Cellular membrane potential and ciliary motility were examined in tissues cultures prepared from frog palate and esophagus epithelia. Addition of micromolar concentrations of extracellular ATP caused membrane hyperpolarization and enhanced the beat frequency. These two effects of ATP were 1) dose dependent, reaching a maximum at 10 microM ATP; 2) dependent on the presence of extracellular Ca2+ or Mg2+; 3) insensitive to inhibitors of voltage-gated calcium channels; 4) abolished after depleting the intracellular Ca2+ stores with thapsigargin; 5) attenuated by quinidine (1 mM), Cs+ (5-20 mM), and replacement of extracellular Na+ by K+; 6) insensitive to charybdotoxin (5-20 nM), TEA (1-20 microM), and apamin (0.1-1 microM); 7) independent of initial membrane potential; and 8) unaffected by amiloride. In addition, extracellular ATP induced an appreciable rise in intracellular Ca2+. Addition of thapsigargin caused an initial enhancement of the ciliary beat frequency and membrane hyperpolarization. These results strongly suggest the involvement of calcium-dependent potassium channels in the response to ATP. The results show that moderate hyperpolarization is closely associated with a sustained enhancement of ciliary beating by extracellular ATP.

Adenosine Triphosphate↗

PubMed1994

Metachronal activity of cultured mucociliary epithelium under normal and stimulated conditions.

In the present work we measured in real time the metachronism and degree of correlation between beating cilia from cultured mucociliary epithelium. The method is based on simultaneous measurement of ciliary beat frequency, phase shifts, and correlation factors in two directions: parallel and perpendicular to the effective stroke direction (ESD). From the phase shifts the lengths of wave components, and consequently the metachronal wavelength and direction, were evaluated. On active ciliary areas of cultured frog esophagus under normal conditions, a relatively high degree of correlation is observed, but cilia are more correlated in direction parallel to ESD which is also the direction of the mucus propulsion. The length of the wave component parallel to ESD is more than twice as large as that of the perpendicular component. The metachronal wavelength was found to be in the range of 5-9 microns, and the direction of the wave propagation was in the range of 90 degrees-125 degrees clockwise to the ESD. When ciliary beat frequency was rapidly increased by extracellular ATP or acetylcholine, only minor effects were observed on the degree of correlation between beating cilia. The length of the wave component parallel to ESD showed the most dramatic effect increasing up to tenfold. The perpendicular to ESD component was not affected by the stimulation. Consequently, the metachronism became more laeoplectic with the angle between the ESD and the wave directions decreasing by 10 degrees-30 degrees, and the metachronal wavelength remained unaltered.

Acetylcholine↗

PubMed1994

Simultaneous measurement of ciliary beating and intracellular calcium.

A novel system for measuring, simultaneously, ciliary beating and intracellular free calcium is presented. The advantages and dynamic nature of the system are demonstrated by measuring the effects of the calcium ionophore lonomycin and of extracellular ATP on ciliated rabbit trachea. The results are discussed with regard to the ciliary and calcium stimulation.

Adenosine Triphosphate↗

PubMed1992

Possible mechanism of ciliary stimulation by extracellular ATP: involvement of calcium-dependent potassium channels and exogenous Ca2+.

Ciliary motility was examined optically in tissue cultures from frog palate epithelium and frog's esophagus as a function of extracellular concentration of adenosine 5'-triphosphate (ATP) and related compounds. The addition of micromolar concentration of ATP caused a strong enhancement of frequency and wave velocity in the direction of the effective stroke. Since adenosine 5'-[beta,gamma imido]-triphosphate (AMP-PNP), a nonhydrolyzable analog of ATP, produces the same effects, ATP hydrolysis is not required. The overall potency is ATP approximately equal to AMP-PNP greater than ADP much greater than adenosine greater than AMP. It is suggested that both the phosphate and the base moieties are involved in ATP binding. The enhancement of ciliary activity by extracellular ATP is dependent on the presence of extracellular Ca2+, which can be replaced by extracellular Mg2+. The effect of a number of potent inhibitors of the voltage-gated calcium channels on the stimulation of ciliary activity by ATP were examined. No effect was detected in the concentration range within which these agents are specific. On the other hand, quinidine, a potent inhibitor of K+ (calcium-dependent) channels, inhibits the effect of ATP. The following model is suggested: exogenous ATP interacts with a membrane receptor in the presence of Ca2+, a cascade of events occurs which mobilizes intracellular calcium, thereby increasing the cytosolic free Ca2+ concentration which consequently opens the calcium-activated K+ channels, which then leads to a change in membrane potential. The ciliary response to these changes is the enhancement of ciliary activity.

Adenosine Triphosphate↗

PubMed1991

Human in vitro nasal and tracheal ciliary beat frequencies: comparison of sampling sites, combined effect of medication, and demographic relationships.

In vitro nasal and tracheal ciliary beat frequencies (CBFs) were measured photometrically in brush samples of 15 patients undergoing bronchoscopy. Tracheal and nasal CBF values in these patients were found to be significantly correlated (r = .74, p less than .01), and the latter did not differ significantly from the nasal CBF of 80 subjects serving as controls. Premedication with morphine and atropine and local anesthesia with lidocaine hydrochloride significantly decreased the CBF. Among the control subjects, a significantly higher CBF (13.6 +/- 1.5 Hz) was found in the young age group (10 to 19 years) as compared to other groups (12.2 +/- 1.7 Hz). Gender and ethnic origin did not influence CBF in any age group. These data support the possible use of the in vitro study of nasal epithelium to reflect the CBF in the lower respiratory tract.

Adult↗

PubMed1990

On metachronism in ciliary systems: a model describing the dependence of the metachronal wave properties on the intrinsic ciliary parameters.

A mathematical model is proposed to explain the dependence of the direction and the length of the metachronal wave on parameters that characterize the ciliary beat, the dimensions of the cilia, and the geometry of their arrangement on the ciliated surface. The metachronal wave is decomposed into two mutually perpendicular components, which are chosen in such a way that the direction of one of them is in the direction of the effective stroke. The magnitudes of the two components are determined by using the concept of the time of delay between adjacent cilia. The properties of the metachronal wave are then calculated as a function of the ciliary parameters. The results obtained with the present model predict that the direction of the wave propagation is strongly dependent on the type of metachronism in the direction of the effective stoke and the polarization in time and in space of the ciliary beat. The metachronal wavelength is found to depend on four parameters: the ciliary length, the angle of the arc projected on the cell surface by the ciliary tip during the recovery stroke, the degree of asymmetry of ciliary beat, and the portion of the cycle occupied by the pause. The metachronal wavelength is also found to be only weakly dependent on the ciliary frequency. At this stage there exists relatively little experimental information with which to characterize fully the metachronal properties of ciliary systems. Even when only partial information exists, the model allows prediction, to within a certain range, of the direction of the wave propagation. It also suggests a possible mechanism for the influence of changes in environmental conditions on wave direction and wavelength. In several cases in which full information does exist, good agreement between the experimental findings and the predictions of the model is found. According to this model it will be worthwhile to invest more effort in measuring the time and space polarization of ciliary beating and times of delay between cilia.

Animals↗

PubMed1990

On self synchronization--aspects of flagellar and ciliary beating.

Mathematical models involving hydrodynamically coupled oscillators are proposed to explain some self-synchronization phenomena observed in flagella and cilia. It is shown using elementary mathematical analysis and computer simulations that the behaviors of the models proposed are similar to those observed in experiment.

Animals↗

PubMed1990

Ciliary activity under normal conditions and under viscous load.

Ciliary metachronism and motility were examined optically in muco-ciliary tissue cultures from three different systems: a) frog's palate epithelium, b) frog's oesophagus, and c) human nasal polyps. In addition, lateral cilia of Mytilus edulis (water transporting cilia) were examined. It was revealed that the degree of synchronization between muco-ciliary systems is lower than that of water transporting cilia. There are no significant differences between different muco-ciliary systems, within the accuracy of our measurement although relatively large statistical ensembles were used. In addition the wavelength and wave direction of the metachronal wave was examined. All four systems exhibit similar wavelength. The metachronal parameters of muco-ciliary systems exhibit fluctuations (as was demonstrated by the degree of synchronization), however, the magnitude and repetitivity of these fluctuations, is dependent on the loading of the ciliary system. We have loaded the system by increasing the viscosity of the medium. Under viscous load the frequency of the beating decreased. The metachronal wavelength became longer and the metachronal coordination type more orthoplectic.

Animals↗