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At least 415 records · Page 23Linked to original sources
PubMed1992

Evidence for an InsP3-gated channel protein in isolated rat olfactory cilia.

Stimulation of rat olfactory cilia (ROC) with odorants leads to a transient elevation in the levels of either cAMP or inositol trisphosphate (InsP3). We have characterized the binding of [3H]InsP3 to isolated ROC. Unlabeled InsP3 displaced [3H]InsP3 binding in a dose-dependent manner (dissociation constant = 3.9 +/- 0.65 microM). Binding was stereospecific and dependent on the number of phosphates in the inositol ring. A ciliary protein of 120 kDa molecular mass was labeled specifically upon exposure of cilia membranes to ultraviolet light in the presence of the 125I-labeled InsP3 analogue 1-O-[N-(4-azidosaliciloxy)-3-aminopropyl-1-phospho]-myo-inositol 4,5-bisphosphate. Labeling of this protein displayed the same stereospecificity as binding of [3H]InsP3 to ROC. In addition, ROC membranes incorporated into a phospholipid bilayer at the tip of a patch pipette displayed an increase in conductance upon exposure to micromolar D-myoinositol 1,4,5-trisphosphate in 45% of the trials (n = 88). The InsP3-gated conductance is relatively nonspecific for cations and is distinct from the cAMP-gated conductance. The conductance displayed stereospecificity consistent with the InsP3 binding experiments. The results suggest that the site of action for odorant-stimulated elevations in InsP3 concentration in rat olfactory cilia is at a ciliary InsP3-gated channel.

Animals↗
PubMed1995

Primary cilia in normal and pathological tissues.

The medical literature in pathology contains a surprising number of anecdotal reports of primary cilia, with authors often expressing some incredulity at finding such organelles. In this update of an earlier review, I will argue that primary cilia should by now be regarded as regular cell organelles, not some form of atypical response to unusual circumstances. In all situations in which their presence is essential, they unquestionably act as sensory transducers. Detection by electron microscopy has been the most reliable means until recently, but is time-consuming and slow for any systematic investigation or experimental approach. Immunostaining with an antibody direct against detyrosinated tubulin is rapidly changing the situation, and we can now detect their presence, frequency, disposition and overall characteristics relatively quickly, allowing better statistical analysis and correlations in abnormal physiological and pathological conditions. To be useful and meaningful, comparative studies need a reliable database of information about primary cilia under 'normal' circumstances. It is hoped that such work will in itself give much further insight into the general significance of these organelles, especially combined with the more experimental approaches that can now be adopted in the study of their development and function, which looks increasingly promising with the new technological improvements.

Animals↗
PubMed1988

Freeze-fracture analysis of the respiratory cilia from the bronchial mucosa of a patient with primary ciliary dyskinesia.

Respiratory cilia of the bronchial mucosa from a 5-year-old boy with clinical evidence of classical Kartagener's syndrome (situs inversus, bronchiectasis and sinusitis) were first examined by means of transmission electron microscopy for identification of the axonemal defects described as typical for primary ciliary dyskinesia (PCD). Additional oscillography was performed on the cilia in vitro, which showed absence of a coordinated ciliary beat frequency. After clear classification of the case as PCD, a freeze-fracture examination of the respiratory cilia was performed, which revealed a higher density of intramembrane particles on the outer fracture face (E-face) than on the inner fracture face (P-face). The results were discussed with regard to probable pathogenetic aspects on PCD.

Biopsy↗
PubMed1980

Kartagener's syndrome with absence of inner dynein arms of respiratory cilia.

Morphologic abnormalities of spermatozoa and respiratory cilia at the electron microscopic level have been described in the immotile-cilia syndrome and chronic respiratory diseases. Most often there has been a total absence of the dynein arms. The current report describes absence of only the inner dynein arm in respiratory cilia from a patient with Kartagener's Syndrome, thus recording another variation in the abnormality of ciliary morphologic features believed to be recessive inherited.

Child↗
PubMed1993

Cyclic AMP-dependent phosphorylation of a 26 kD axonemal protein in ovine cilia isolated from small tissue pieces.

To study cyclic adenosine monophosphate (cAMP)-dependent phosphorylation events in ovine cilia in vitro, we adapted published axonemal isolation methods to obtain pure mammalian axonemal proteins from small ovine tracheal mucosa pieces with a surface area of only 1 cm2. The isolated axonemes could be reactivated in vitro upon ATP addition, thereby attesting to their functional integrity. The axonemal protein yield from these small mucosa pieces was high enough to allow protein concentration measurements of each sample and axonemal polypeptide analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). cAMP is known to increase ciliary beat frequency, possibly through a phosphorylation event in the axoneme. To study cAMP-dependent phosphorylation events in ovine tracheal cilia, these axonemal preparations were exposed to [gamma-32P]ATP under conditions that stimulated or inhibited kinase activity. Analysis of axonemal polypeptides by SDS-PAGE and subsequent autoradiography showed that an axonemal protein with a M(r) of 26 kD is the only polypeptide consistently phosphorylated in a cAMP-dependent manner. The phosphorylation of this protein could be diminished by a highly specific inhibitor of cAMP-dependent protein kinase, KT-5720. The addition of calcium did not affect label incorporation into this protein during cAMP treatment. In the presence of cAMP and calcium, inhibitors of protein kinase C and calcium/calmodulin-dependent kinase did not change the level of phosphorylation of the 26 kD protein. We conclude that cAMP treatment of isolated mammalian cilia results in the phosphorylation of a single protein with a M(r) of 26 kD (p26).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1996

Morphologic alterations in the cilia of a cat.

Based upon ultrastructural findings and computed tomography, a presumptive diagnosis of feline primary ciliary dyskinesia was made in a 2.5-year-old cat. The cat demonstrated morphologic alterations in the ultrastructure of oviductal cilia. In the oviduct, axonemal abnormalities were detected in 132 (20%) of 660 cross-sectioned cilia. The main ultrastructural ciliary defects were a lack of central microtubules, transposition of peripheral doublets to the center of the axoneme, supernumerary microtubules, and morphologic abnormalities of peripheral doublets. Computed tomography of the chest was consistent with early lesions of bronchiectasis, suggesting chronic stagnation of respiratory secretions, attributed to abnormal function of respiratory cilia. Specifically, the cranial branches of the cat's bronchi were wider and thicker than those of five healthy controls. Foci of pleural thickening and interstitial enlargement were also observed.

Animals↗
PubMed1988

Immunoelectron microscopic demonstration of S-100b protein-like in centriole, cilia, and basal body.

We report here the ultrastructural localization of S-100b protein-like immunoreactivity in the centriole, cilia, and basal body. Duodenum and trachea of guinea pigs and rats were fixed and immunostained by the protein A-gold method. All centrioles, cilia, and basal bodies observed showed clear S-100b protein-like immunoreactivity. Specific colloidal gold particles were located over the microtubules in these cell organelles. However, other microtubules scattered throughout the cytoplasm were devoid of immunoreactivity. Although the functional significance of S-100b protein-like immunoreactivity in the centriole, cilia, and basal bodies remains to be elucidated, the present results introduce new perspectives into the investigation of localization and function of S-100 proteins.

Animals↗
PubMed1992

Dynein from serotonin-activated cilia and flagella: extraction characteristics and distinct sites for cAMP-dependent protein phosphorylation.

Serotonin, an activator of adenylate cyclase, stimulates motility in molluscan gill cilia and sperm flagella. To determine and compare potential targets of cAMP action, dynein was prepared from the lateral gill.cilia and sperm flagella of the mussel Mytilus edulis and the clam Spisula solidissima. In the flagella of both species, high-salt extraction removes about half of the ATPase activity, half of the alpha and beta heavy chains, and the outer arms. The dynein from both species sediments at 18-20 S, contains two or three intermediate chains, and three light chains. High-salt plus detergent removes most of the remaining dynein ATPase, alpha and beta heavy chains, and inner arms, also yielding a stable 18-20 S particle. In gill cilia of both species, high-salt extraction removes only 12-18% of the ATPase, up to 1/3 of the alpha heavy chains, an equivalent amount of beta heavy chain, and a subset of the outer arms. The dynein sediments at 18-20 S and, in Spisula, the heavy, intermediate, and light chains precisely co-sediment. High-salt plus detergent removes another 1/3 of the alpha heavy chains, an equivalent amount of beta heavy chain, and the remaining outer arms. The ATPase sediments mainly as a 13-14 S form showing considerable dissociation of co-sedimenting intermediate and light chains. The inner arms and at least half of the ciliary dynein ATPase activity remain unextractable, corresponding in mass mainly to an apparent beta heavy chain that is vanadate-cleavable. Cyclic AMP-dependent, calcium-independent phosphorylation takes place on specific dynein light chains in cilia but on only the dynein alpha heavy chain in flagella. Pre-activation of the flagella prevents subsequent addition of labeled phosphate. Phosphorylation has no effect on the steady-state ATPase properties. The single phosphate added to the flagellar alpha chain is located within the LUV1 vanadate photocleavage fragment. Considering the probable locus of the light chains and the site of the alpha heavy chain phosphorylation, both beyond the active site and toward the base of the molecule, these distinct phosphorylations may regulate dynein action by modulating arm flexibility or interaction.

Adenosine Triphosphate↗
PubMed1984

A neurofibrillar method stains solitary (primary) cilia in the mammalian retina: their distribution and age-related changes.

Richardson's reduced silver method, developed for the staining of autonomic nerve fibres in the mammalian intestine, is shown consistently to stain solitary (primary) cilia and diplosomes of the cells of the retinae of cats and rabbits. The cilia comprise a centriole, a basal body and an axoneme with a 9 + 0 complement of microtubules. Probably all retinal cells possess a cilium during their development but, contrary to previous reports, not all retain the axoneme after birth. Axonemes are absent from horizontal, bipolar, microglia, Müllerian, and probably some other glial, cells; all of which showed paired centrioles (diplosomes) after staining. Photoreceptor, amacrine, interplexiform, displaced amacrine and ganglion cells have each one cilium. These differences between cell types persist, without significant change, in the retina of adult rabbits up to the age of 2 years, and in the cat up to 5 years. The alpha-type ganglion cells of the cat are an exception. In 4 to 8-week-old kittens they are all ciliated, like other types of ganglion cells. But by two years about 30% of central area alpha-cells lack an axoneme. Individual cells may have only diplosomes, unusual dispositions of the centrioles in the perikaryal cytoplasm, or even show complete loss of the whole ciliary apparatus. By 5 years of age the proportion of those alpha-cells showing unusual arrangements has increased to approximately 70%, while less than 5% of the other types of ganglion cells are so affected. Cilia of peripheral alpha-cells change at a different rate and by 5 years of age are approximately like the 2-year-old central area population.

Aging↗
PubMed2005

De novo formation of left-right asymmetry by posterior tilt of nodal cilia.

In the developing mouse embryo, leftward fluid flow on the ventral side of the node determines left-right (L-R) asymmetry. However, the mechanism by which the rotational movement of node cilia can generate a unidirectional flow remains hypothetical. Here we have addressed this question by motion and morphological analyses of the node cilia and by fluid dynamic model experiments. We found that the cilia stand, not perpendicular to the node surface, but tilted posteriorly. We further confirmed that such posterior tilt can produce leftward flow in model experiments. These results strongly suggest that L-R asymmetry is not the descendant of pre-existing L-R asymmetry within each cell but is generated de novo by combining three sources of spatial information: antero-posterior and dorso-ventral axes, and the chirality of ciliary movement.

Animals↗
PubMed2005

Nodal cilia dynamics and the specification of the left/right axis in early vertebrate embryo development.

Nodal cilia dynamics is a key factor for left/right axis determination in mouse embryos through the induction of a leftward fluid flow. So far it has not been clearly established how such dynamics is able to induce the asymmetric leftward flow within the node. Herein we propose that an asymmetric two-phase nonplanar beating cilia dynamics that involves the bending of the ciliar axoneme is responsible for the leftward fluid flow. We support our proposal with a host of hydrodynamic arguments, in silico experiments and in vivo video microscopy data in wild-type embryos and inv mutants. Our phenomenological modeling approach underscores how the asymmetry and speed of the flow depends on different relevant parameters. In addition, we discuss how the combination of internal and external mechanisms might cause the two-phase beating cilia dynamics.

Amniotic Fluid↗
PubMed2006

Clustering of cyclic-nucleotide-gated channels in olfactory cilia.

Olfactory cilia contain the known components of olfactory signal transduction, including a high density of cyclic-nucleotide-gated (CNG) channels. CNG channels play an important role in mediating odor detection. The channels are activated by cAMP, which is formed by a G-protein-coupled transduction cascade. Frog olfactory cilia are 25-200 microm in length, so the spatial distribution of CNG channels along the length should be important in determining the sensitivity of odor detection. We have recorded from excised cilia and modeled diffusion of cAMP into a cilium to determine the spatial distribution of the CNG channels along the ciliary length. The proximal segment, which in frog is the first 20% of the cilium, appears to express a small fraction of the CNG channels, whereas the distal segment contains the majority, mostly clustered in one region.

Animals↗
PubMed1982

Electron microscopic observations on single cilia in the intrahepatic biliary ducts in some birds.

The intrahepatic biliary passage in five species of birds was investigated with the transmission electron microscope. The avian liver was characterized by a frequent occurrence of intralobular bile ductules and canaliculo-ductular junctions in the parenchyma. It was further characterized by solitary bile ductular epithelial cells intercalated among hepatocytes surrounding bile canaliculi. The present study first revealed that avian bile ductular epithelial cells possess a long single cilium. Its basal body (distal centriole) was connected to a basal foot and slender rootlet and accompanied by a proximal centriole. The hepatocytes facing the bile passage possessed no cilium, although they frequently had a diplosome in their apical cytoplasm. The single cilia of the bile ductular epithelium gradually tapered toward the tip. The original fiber pattern in the most proximal part was peripheral 9 doublets +0. In the ciliary shaft, the doublets altered into singlets which were diminished in number gradually toward the distal parts of the shaft, so that in the tip only one singlet remained. Since these fiber patterns in the single cilia markedly deviated from the 9 + 2 fiber pattern of the ordinary motile cilia, they may not be motile, but properly regarded as sensory or chemoreceptors.

Animals↗
PubMed1993

Ultrastructure and function of cilia and spermatozoa flagella in a patient with Kartagener's syndrome.

A 37-year-old man with the characteristic clinical signs of Kartagener's syndrome, including bronchiectasia, chronic paranasal sinusitis, and situs inversus, was found to have immotile cilia and sperm. Electron microscopy demonstrated that the majority of cilia lacked dynein arms and radial spokes and that various defects of microorgans existed in the sperm. The most frequent defect was total defect of axoneme followed by defect of inner dynein arms in the sperm. These findings suggest an association between the structural abnormality of absent inner dynein arms and immotility of cilia and sperm in Kartagener's syndrome.

Adult↗
PubMed1995

Single cilia in human epidermis are susceptible to challenge.

Single cilia were found in melanocytes and basal keratinocytes in normal epidermis and in epidermis subjected to various types of exposure. Quantitative electron microscopy showed that about 31% of the melanocytes and 71% of the keratinocytes from normal skin possessed a ciliary apparatus, but there was a wide individual variation. A statistically significant decrease in the number of ciliated keratinocytes followed exposure to nickel, sodium lauryl sulphate, UVA and UVB irradiation. There was no statistically significant difference between controls and skin subjected to various exposures regarding the number of ciliated melanocytes. We have earlier reported that single cilia are frequently seen on epithelial cells capable of mitotic activity in human oral and vaginal epithelia. The present study showed that single cilia react to external exposures. The cilium might be a sensitive, easily damaged organelle, or the decrease in ciliated cells might reflect a change in normal mitotic activity as a result of exposure.

Cilia↗
PubMedDate not supplied

Otologic manifestations of the immotile-cilia syndrome.

The immotile-cilia syndrome is caused by a hereditary, inborn, ultrastructural defect of the cilia, rendering them immotile or poorly motile and thereby abolishing mucociliary clearance. Five cases are presented and the otologic manifestations are described in some detail. The syndrome should be suspected in children who have a persistent secretory otitis media with recurring bouts of acute otitis media, and a perpetual cough with repeated episodes of bronchitis. A lobar atelectasis is a frequent finding. Half the cases also have situs inversus. The immotile-cilia syndrome is of special interest to the otologist, as it seems to throw some light on the pathogenesis of secretory otitis media.

Adolescent↗
PubMed1995

The influence of specimen preparation on artefacts in scanning electron microscopy of respiratory cilia.

The effects of various handling procedures used in preparing specimens of human and pig respiratory mucosa for scanning electron microscopy (SEM) were studied. Using five different washing methods, the percentage area of mucosa covered with extracellular material varied from 1.5 to 53.1%. The best results were achieved when specimens were washed by gently inverting a sample 30 times in a container filled with physiological saline. Fixation and drying of the surface layer caused disorientation of cilia and made examination difficult. Mechanical damage caused loss of cilia and rupture of the cell membrane. For SEM of respiratory cilia it is important to wash the specimen in saline before fixation and to use biopsy forceps as little as possible.

Animals↗
PubMed2001

[Observation of nasal mucosal cilia ultrastructure of Kartagener's syndrome].

OBJECTIVE: To report the ultrastructure of nasal mucosal cilia of Kartagener's syndrome and the operative results of 2 cases. METHODS: Both two cases were underwent endoscopic sinus surgery. Transmission and scanning electron microscopic examinations of nasal mucosa for case 1 were performed. RESULTS: All 2 cases presented situs inversus, chronic paranasal sinusitis and bronchiectasis. The electron microscopic examination showed that the shape of nasal cilia and "9 + 2" structure were normal, but the lack of inner dynein arm of cilia and confused arrangement of central pair of microtubules were confirmed. The recurrent of nasal polyps after 1-year follow-up was observed in case 1. CONCLUSIONS: The disturbance of mucociliary clearance due to congenital ciliary structure defect might be the cause of chronic sinusitis and polyps, which might be one of reasons for poor prognosis after operation and we should pay more close attention.

Adolescent↗