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PubMedDate not supplied

The suprahabenular recess in the rat: a quantitative study of ciliated cells, supraependymal cells and some specific features of supraependymal fibres.

Using light microscopy, scanning and transmission electron microscopy, an extensive quantitative analysis of ciliated ependymal cells, supraependymal cells and supraependymal fibres in the suprahabenular recess of twenty-two adult Wistar rats was carried out. In addition, six animals were used to determine the origin of supraependymal fibres and their relationship to ependyma. The findings were as follows: ciliated cells-mean 3.32 +/- 0.26 per 1000 micron 2 (range 0-10); supraependymal cells-5.91 +/- 1.37 per recess (range 2-16). Average number of cilia per cell in different animals ranged from 31 to 33. Sex did not influence the density of ciliated ependyma or supraependymal cells. Evidence was obtained that some supraependymal fibres are closely associated with aberrant commissural fibres. The supraependymal fibres enter the suprahabenular recess and form gap junctions, and other types of junctions, with subjacent ependyma.

Animals↗
PubMed1985

Immuno-electron-microscopic localization of a centriole-related antigen in ciliated cells.

An antigen common to purported centriolar and basal body regions of a variety of cell types was previously visualized by immuno-fluorescence microscopy. The present study demonstrates the localization of the antigen relative to the defined basal body structures of ciliated tracheal cells at the electron-microscopic level. After ethyldimethylaminopropyl carbodiimide-glutaraldehyde-saponin (EGS) fixation and permeabilization, immunoferritin labeling is consistently found associated with amorphous electron-opaque material in proximity to basal bodies and their ciliary rootlets, but not with basal body microtubules themselves. This distribution pattern is distinct from that of other proteins found in the apical region of ciliated cells, such as calmodulin. It is proposed that the dense material may be analogous to pericentriolar material of centrosomes.

Animals↗
PubMed1986

Developmental potential of ciliated cells of ceractinomorph sponge larvae.

The fate of the ciliated epithelium of sponge larvae at metamorphosis has been the subject of debate for many years. In order to resolve the phylogenetic position of the Porifera it must be determined whether these cells migrate inwards to become adult choanocytes or are lost at metamorphosis. We report an approach to this problem by observing metamorphosing larvae using electron microscopy and by assaying the aggregation potential of ciliated and central larval cells fractionated on Percoll gradients. Our results indicate that larval epithelial cells are phagocytosed during metamorphosis and that they do not have aggregation capability. The latter is essential to permit choanocyte chamber formation. Larval archaeocytes do possess aggregation capability. We argue that larval ciliated cells are dispersal effectors only and are terminally differentiated. Thus 'inversion of layers' as claimed by earlier authors does not occur. These results are discussed in relation to the role of aggregation facilitating systems and to the structure of sponge larvae.

Animals↗
PubMed1976

[Histochemical and histological investigations on the human fallopian tube under different hormonal influences. I. Demonstration of ATPase with special reference to reactive ciliated cells (author's transl)].

The localization of the Mg++-activated adenosine triphosphatase (ATPase) in the human Fallopian tube has been studied by means of histochemical methods. The samples were obtained from 18 women in the age from 23--62 years. Some of them were treated by various steroid hormones. Endosalpinx ciliary ATPase-activity represents dynein and is therefore an indicator of ciliary motility. Estrogens and gestagens have a different influence on the ATPase-activity. All cilia of one ciliated cell react in the same manner and may be regarded as a reaction unit. The relation of negative to positive ciliary borders differs characteristically in the tubal isthmus, ampulls and infundibulum and coincides with commonly known phenomena of egg transport through the oviduct. Postovulatory, reaction units increase in ampulla and infundibulum compared with the proliferative phase. The oviducts of postmenopausal women possess but a scanty outfit of reaction units. Short-time treatment with estrogen in the early secretory phase results in a great number of reaction units in all tubal segments; a similar treatment in the proliferative phase diminishes the reaction units in the ampulla. Midcycle progesterone treatment activates the ciliary ATPase in the isthmus. Low doses of lynestrenol (minipill) in the proliferative phase leads to a decrease of reaction units in all tubal segments; the pattern of ciliary reaction under low doses of lynestrenol at the time of ovulation coincides with that of the proliferative phase. Treatment with a contraceptive steroid (0,05 mg ethinylestradiol and 0,25 d-norgestrel) causes a considerable activation of the ciliary ATPase in all portions of the oviduct.

Adenosine Triphosphatases↗
PubMed1988

Response of nasal ciliated cells of the guinea-pig during allergic reactions in human blood.

Nasal mucosal samples from thirty-five guinea-pigs were placed in a chamber, containing the medium RPMI 1640. Ciliary activity (beats.s-1) of the most active cell in each mucosal culture was measured using a photoelectric method. The RPMI 1640 was then replaced by 2 ml of RPMI 1640 and 1 ml of heparinized human blood from a non-allergic patient, with a ragweed-sensitive nasal allergy, or patients with D. farinae-sensitive nasal allergy: the ciliary activity of the same ciliated cell in each culture did not change significantly. We then added 1 micrograms of D. farinae extracts to evoke an in vitro allergic reaction. Ciliary excitation was induced when D. farinae extracts were added to the blood from D. farinae-sensitive subjects but not when added to blood from subjects without sensitivity to D. farinae. The peak and plateau of such an event occurred after 20-30 min. Ciliary stimulations were correlated with D. farinae-induced histamine release from whole blood. Thus, in vitro allergic reactions stimulate activity in cilia from normal nasal mucosa.

Allergens↗
PubMed2005

InsP3 signaling induces pulse-modulated Ca2+ signals in the nucleus of airway epithelial ciliated cells.

The phenomenology of nuclear Ca(2+) dynamics has experienced important progress revealing the broad range of cellular processes that it regulates. Although several agonists can mobilize Ca(2+) from storage in the nuclear envelope (NE) to the intranuclear compartment (INC), the mechanisms of Ca(2+) signaling in the nucleus still remain uncertain. Here we report that the NE/INC complex can function as an inositol-1,4,5-trisphosphate (InsP(3))-controlled Ca(2+) oscillator. Thin optical sectioning combined with fluorescent labeling of Ca(2+) probes show in cultured airway epithelial ciliated cells that ATP can trigger periodic oscillations of Ca(2+) in the NE ([Ca(2+)](NE)) and corresponding pulses of Ca(2+) release to the INC. Identical results were obtained in InsP(3)-stimulated isolated nuclei of these cells. Our data show that [Ca(2+)](NE) oscillations and Ca(2+) release to the INC result from the interplay between the Ca(2+)/K(+) ion-exchange properties of the intralumenal polyanionic matrix of the NE and two Ca(2+)-sensitive ion channels-an InsP(3)-receptor-Ca(2+) channel and an apamin-sensitive K(+) channel. A similar Ca(2+) signaling system operating under the same functional protocol and molecular hardware controls Ca(2+) oscillations and release in/to the endoplasmic reticulum/cytosol and in/to the granule/cytosol complexes in airway and mast cells. These observations suggest that these intracellular organelles share a remarkably conserved mechanism of InsP(3)-controlled frequency-encoded Ca(2+) signaling.

Animals↗
PubMed2005

Chloride transport in nasal ciliated cells of cystic fibrosis heterozygotes.

Studying subjects heterozygous for mutations of the cystic fibrosis (CF) gene may help clarify the impact on disease onset of CF transmembrane conductance regulator protein (CFTR-)-dependent chloride secretion. CFTR-mediated chloride transport was evaluated in 52 heterozygous subjects, 32 healthy control subjects, and 77 patients with CF with class I or II mutations. We measured the change in nasal potential difference in response to chloride-free isoproterenol solution for each subject and used a video-imaging fluorescent dye assay to assess the percentage of nasal ciliated cells with cAMP-dependent anion conductance. Our findings did not confirm the standard assumption that heterozygosity implies 50% of normal CFTR function. Half the heterozygous subjects had CFTR-mediated chloride transport levels below 50% of the normal range, and one-third had levels similar to those of the patients with CF. This reduced CFTR function was not associated with an elevated prevalence of CF-like symptoms in heterozygous subjects but was highly related to respiratory status in the patients with CF. These data suggest that CFTR-dependent chloride conductance does not directly modulate disease severity but may be part of a more global defect in patients with CF involving other CFTR functions or currently unknown modulatory factors.

Chlorides↗
PubMed1982

Immune localization of calmodulin in the ciliated cells of hamster tracheal epithelium.

Melachronous beating of cilia of epithelial surfaces of most respiratory airways moves the overlying mucous layer in a caudal direction. The molecular mechanisms controlling ciliary beat remain largely unknown. Calcium, an element in its cationic form, is ubiquitous in biological functions and its concentration is critical for ciliary beating. Calmodulin, a calcium-binding protein which regulates the activity of many enzymes and cellular processes, may regulate ciliary beating by controlling enzymes responsible for mechanochemical movement between adjacent peripheral microtubule doublets composing the ciliary axoneme. As a first step in describing a calmodulin-related controlling mechanism for ciliary beating, calmodulin was localized in the ciliated cells lining the respiratory tracts of hamsters by electron microscopy, using an indirect immunoperoxidase technique with anticalmodulin antibodies as the molecular probe. Thin-sections revealed calmodulin located on microtubules and dynein arms of the ciliary shaft, basal body, apical cytoskeletal microtubules, and plasma membranes in specimens fixed with 1 mM Ca+2. Specimens fixed with less Ca+2 (1 microM), Mn+2, Mg+2, and EGTA showed a diffuse pattern of calmodulin with loci of greatest densities on basal body microtubule triplets. Demembranated specimens showed a less specific localization on axonemal microtubules but only on cells fixed with Ca+2. Calmodulin, by binding calcium, may function in ciliary beating in the respiratory tract of mammals either directly or indirectly through its effects on the energy-producing enzymes and by control of Ca+2 flux through plasma membranes.

Animals↗
PubMed1986

Ciliated cells of the trachea of the rabbit, treated with cis-diamminedichloroplatinum (II) alone, or in combination with ionizing radiation.

The ciliated epithelium of the rabbit trachea was irradiated with daily fractions of 2 Gy to an accumulated dose of 20 Gy (TD: 2, 6, 10, 16, or 20 Gy). Fifteen to forty-five minutes before start of the first irradiation (treatment day 1), 5 mg cis-DDP was given by intraperitoneal injection to each rabbit. Examination was made 1 - 10 days after each fractionation schedule, when specimens were taken for investigations. Scanning electron microscope investigations showed a gradual development of ciliary damage from blebs on the cilia to swollen tips, broken and bent cilia and finally an epithelial lining with areas free from cilia with a surface covered with microvilli-like structures. SEM also showed cell loss, and remnants of dead cells on the surface together with detritus. By transmission electron microscope ciliary damage, cell death and cell loss of the ciliated cell layer as well as exfoliation of portions of goblet-like cells on the surface could be confirmed. The irradiated ciliated epithelium and the untreated control epithelium in each animal showed no difference in this respect. Thus no enhancement of the effects of radiation could be observed. The development of ultrastructural damage may be due to a cytotoxic effect of the drug on the ciliated epithelium. However, 19 days after the start of cis - DDP injection, a hyperplasia of the basal cell layer was observed, which indicates that the observed cytotoxicity of the drug is reversible and a normalisation occurs during the last days of observation in this study.

Animals↗
PubMed2005

Arrest in ciliated cell expansion on the bronchial lining of adult rats caused by chronic exposure to industrial noise.

Workers chronically exposed to high-intensity/low-frequency noise at textile plants show increased frequency of respiratory infections. This phenomenon prompted the herein investigation on the cytology of the bronchial epithelium of Wistar rats submitted to textile noise. Workplace noise from a cotton-mill room of a textile factory was recorded and reproduced in a sound-insulated animal room. The Wistar rats were submitted to a weekly schedule of noise treatment that was similar to that of the textile workers (8h/day, 5 days/week). Scanning electron microscopy (SEM) was used to compare the fine morphology of the inner surface of the bronchi in noise-exposed and control rats. SEM quantitative cytology revealed that exposure to noise for 5-7 months caused inhibition in the natural expansion of the area occupied by ciliated cells on the bronchial epithelium as adult rats grow older. This difference between noise-exposed and age-matched control rats was statistically significant (P<0.05) and documents that the cytology of the rat bronchial epithelium is mildly altered by noise exposure. The decrease in the area of bronchial cilia may impair the mucociliar clearance of the respiratory airways and, thus, increase vulnerability to respiratory infection.

Animals↗
PubMed1998

Induction of a differentiated ciliated cell phenotype in primary cultures of Fallopian tube epithelium.

Human Fallopian tubal epithelial cells in culture lose morphological features associated with the epithelium in situ and the extent to which they retain their in-vivo phenotype or function is unknown. In order to address this question, immunocytochemical markers were identified which distinguish secretory (HMFG2+, LhS28-) from ciliated (HMFG2-, LhS28+) epithelial cells in tissue sections of Fallopian tube. These markers were used to analyse the phenotype of tubal cells in vitro. Primary cultures of human tubal epithelial cells were seeded onto glass and grown to confluence before addition of oestradiol-17beta. In the absence of hormone, tubal epithelial cells expressed cytokeratins and nuclear receptors for oestrogen and progesterone and adopted a homogeneous (HMFG2+, LhS28-) secretory cell phenotype. Following the addition of oestradiol-17beta, a proportion of cells became positive for LhS28. The induction of a ciliated epithelial cell phenotype was confirmed by scanning electron microscopy, where on permeable collagen membranes, approximately one-third of tubal epithelial cells became ciliated in the presence of oestradiol-17beta. We suggest that in vitro, tubal epithelial cells adopt an immature secretory-like phenotype and that oestrogen can induce differentiation to a ciliated epithelial cell phenotype.

Antibodies, Monoclonal↗
PubMed2005

Severe acute respiratory syndrome coronavirus infection of human ciliated airway epithelia: role of ciliated cells in viral spread in the conducting airways of the lungs.

Severe acute respiratory syndrome coronavirus (SARS-CoV) emerged in 2002 as an important cause of severe lower respiratory tract infection in humans, and in vitro models of the lung are needed to elucidate cellular targets and the consequences of viral infection. The SARS-CoV receptor, human angiotensin 1-converting enzyme 2 (hACE2), was detected in ciliated airway epithelial cells of human airway tissues derived from nasal or tracheobronchial regions, suggesting that SARS-CoV may infect the proximal airways. To assess infectivity in an in vitro model of human ciliated airway epithelia (HAE) derived from nasal and tracheobronchial airway regions, we generated recombinant SARS-CoV by deletion of open reading frame 7a/7b (ORF7a/7b) and insertion of the green fluorescent protein (GFP), resulting in SARS-CoV GFP. SARS-CoV GFP replicated to titers similar to those of wild-type viruses in cell lines. SARS-CoV specifically infected HAE via the apical surface and replicated to titers of 10(7) PFU/ml by 48 h postinfection. Polyclonal antisera directed against hACE2 blocked virus infection and replication, suggesting that hACE2 is the primary receptor for SARS-CoV infection of HAE. SARS-CoV structural proteins and virions localized to ciliated epithelial cells. Infection was highly cytolytic, as infected ciliated cells were necrotic and shed over time onto the luminal surface of the epithelium. SARS-CoV GFP also replicated to a lesser extent in ciliated cell cultures derived from hamster or rhesus monkey airways. Efficient SARS-CoV infection of ciliated cells in HAE provides a useful in vitro model of human lung origin to study characteristics of SARS-CoV replication and pathogenesis.

Angiotensin-Converting Enzyme 2↗
PubMed1983

Scanning electron microscopy and recording of the physiological activity of tracheal ciliated cells treated by fractionated irradiation.

The ciliated epithelium of the rabbit's trachea was irradiated with daily fractions of 2 Gy to an accumulated dose of 20 Gy. The beat frequency of the cilia was recorded and specimens were taken for SEM-, TEM- and LM-investigations. Examination was made 1-10 days after each fractionation schedule. An increased ciliary beat frequency was recorded at 2 Gy and 4 Gy. With increasing dose, there was an inverted relationship to the frequency. Light-microscopy showed edema and an increased amount of goblet cells in relation to the increasing dose. With SEM an increased number of ciliary blebs could be seen. These could be classified according to size and number, and showed a positive correlation to the dose. TEM-investigations showed signs of increased intracellular activity at higher doses in the form of multilobulated nuclei and an increasing number of nuclear pores. At lower doses, an increased amount of mitochondria appeared in the apical part of the cell. It is at present difficult to evaluate any correlation between the physiological activity and the morphology. More biological data are needed to explain the early irradiation effects.

Animals↗
PubMed2000

Regulation of a hyperpolarization-activated chloride current in murine respiratory ciliated cells.

1. The properties of a hyperpolarization-activated Cl- current (Ihyp-act) in murine ciliated respiratory cells have been studied using whole cell patch clamping. 2. The current-voltage relationship was inwardly rectifying which was due to voltage-dependent gating of the channel. 3. Inward current was markedly sensitive to the extracellular Cl- concentration, an effect that was not related to changes in transmembrane Cl- gradient. Decreasing extracellular Cl- concentration to 6 mM caused a 70 % reduction in inward current with the dose-response relationship exhibiting a Hill coefficient of approximately 2.0 and an IC50 of 29 mM. 4. External anion replacement gave a selectivity sequence of Br- >= I- > Cl- > gluconate = aspartate. The more permeant halides significantly increased current density while the less permeant anions decreased current density, indicating that an extracellular anion is important for channel activity. 5. The conductance was unaffected by exposure to anisotonic pipette solutions or to increases in intracellular cAMP; however, current density was reduced dose dependently by increases in intracellular calcium concentration from 0.1 to 0.5 microM. These results indicate that Ihyp-act is unlikely to be involved in either volume regulation or cAMP/Ca2+-stimulated fluid secretion. 6. Decreasing extracellular pH to 5.0 irreversibly inhibited Ihyp-act. However, the current was fully active over the pH range 5.4-9.0 making it unlikely that it is modulated by extracellular pH under physiological conditions. 7. We speculate that Ihyp-act may have a role in basal Cl- absorption, acting as a Cl- sensor to maintain optimal volume and composition of airway surface liquid.

Adenosine Triphosphate↗
PubMed1983

Improved method for transmission electron microscopy of ciliated cell monolayers maintained on gas-permeable membranes.

A new method is described for preparing thin sections for transmission electron microscopy of ciliated respiratory epithelial cells cultivated in vitro on Teflon membranes. Hamster tracheal explant cultures were treated with collagenase to promote epithelial migration. Patches of monolayer growth occurred in the vicinity of the explants when the latter were incubated on FEP-etched Teflon membranes in Chamber/Dishes. Cells in the monolayer maintained a high degree of differentiation as evidenced by active ciliary motion. The monolayer was fixed and processed in situ. During the embedding process the chamber was supported on a solid Teflon cylinder to keep the membrane taut as it was heated to polymerize the resin. The membrane was removed and the monolayered cells were re-embedded and sectioned. Electron microscopic examination of cultures revealed clear cell profiles on a flat, even basal plane. This method provides for a normal spatial configuration of cells in a monolayer format, and should prove useful for electron microscopy studies of any cells cultured on flexible films.

Animals↗
PubMedDate not supplied

[Epidermal growth factor stimulates proliferation and DNA synthesis in ciliate cells].

We studied the effect of murine epidermal growth factor on cell proliferation and DNA synthesis in macronuclei of ciliate Tetrahymena pyriformis G1. Mitogenic effect of epidermal growth factor on proliferation-induced tetrahymena cells has been revealed. This effect is due to the induced progression of cells at G1 and, consequently, their earlier entering DNA synthesis phase of the first cell cycle. Epidermal growth factor had no mitogenic effect on the resting cells from stationary culture (G0 phase) whose development is independent of the growth factors in the medium.

Animals↗
PubMed1995

The effects of benomyl and its breakdown products carbendazim and butyl isocyanate on the structure and function of tracheal ciliated cells.

The effects of the fungicide benomyl and its breakdown products, carbendazim and butyl isocyanate, were examined on canine tracheal epithelial tissue in primary culture. Changes in ciliary frequencies were monitored with an optical spectrum analysis system. Serial dilutions of the test compounds were prepared in 100% corn oil and applied to the cell cultures for intervals up to 6 hours and frequencies measured at intervals of 15 minutes to 1 hour. Benomyl and butyl isocyanate caused concentration-dependent decreases in ciliary beat frequency. Benomyl at 300 micrograms/ml (3 mM) caused ciliostasis within 75 minutes of exposure. Butyl isocyanate at a molar concentration three times lower than benomyl (1 mM) caused a similar response, although within 30 minutes. The IBC50 for benomyl was 0.75 mM, while for butyl isocyanate it was 0.52 mM. Carbendazim caused a moderate decrease in frequency over a 6 hour exposure period. Benomyl caused moderate to severe swelling of the mitochondria of ciliated epithelial cells with other cell organelles appearing normal. Butyl isocyanate did not cause any noticeable effect on cell ultrastructure and the apparently low rate of penetration of carbendazim into cells made it impossible to obtain an effect which justified ultrastructural analysis. It appears, at least for benomyl and butyl isocyanate, that while the physiological effect of these two compounds (inhibition of ciliary beat) is the same, the sites of action in the cell may be different.

Animals↗
PubMed1993

Secretory proteins in a ciliate cell: identification of epitopes common to numerous polypeptides.

Secretory vesicles of the ciliate Pseudomicrothorax dubius, called trichocysts, are separated into > 40 proteins by two-dimensional gel electrophoresis. The trichocyst, composed of a shaft and four arms, is in a condensed state when docked in the cell cortex, and it elongates into an extended state during exocytosis. Monoclonal antibodies (mAbs) were raised against trichocyst proteins. Their reactivities were analysed: 1) on Western blots of extended, isolated trichocysts by immunolabeling; 2) on entire cells and extended trichocysts by indirect immunofluorescent binding assay (IFA); 3) on semi-thin sectioned cells by IFA; and 4) on ultra-thin sections of cells by immunogold labeling. mAb IV 4E5 labels major trichocyst proteins at 15-19, 22 and 24 kDa, pI 4.6-6.6. The epitope recognized by mAb IV 4E5 is common to as many as 30 proteins and suggests a family of proteins with possible sequence homology. By IFA, the shafts of extended trichocysts are labeled. The shafts of condensed trichocysts are labeled on both semi-thin sections in Lowicryl and ultrathin sections. On semi-thin Epon sections, the part of the trichocyst which is labeled is arm-like. mAb VI 2D12 labels three major trichocyst proteins at 31 kDa, pI 5.0-5.4. The arms of extended trichocysts are labeled by IFA, but are only weakly labeled on ultrathin sections. The shaft of extended trichocysts is labeled by IFA, and the shaft of condensed trichocysts is labeled on ultrathin sections.

Animals↗