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PubMed1970

Cilia and vesicular particles in the endocrine pancreas of the Mongolian gerbil.

Though the general appearance and the cellular composition of the pancreatic islets of the Mongolian gerbil (Meriones unguiculatus) conformed to those of most other rodent species, some peculiar ultrastructural details were found. Thus there were diversiform, mainly vesicular particles with varying electron opacity in these islets. The vesicular particles showed a clear association to cilia which seemed to possess a basic fiber pattern of 9 + 0. The basal bodies were localized in the cytoplasm of the islet parenchymal cells, most often in the beta-cells, and the vesicular particles occurred in the portions of cilia that were protruding into intercellular spaces. The cilia were often swollen, and the vesicular particles were mainly found in the space between the ciliary membranes and the longitudinal fibers. A few vesicular particles could be seen inside and sometimes seemingly in contact with these fibers. Occasionally, there were morphologically similar structures in the cytoplasm of adjacent beta-cells. The vesicular particles were differentiated from the vesicles occurring in nerve structures by their larger size, as well as by their heterogeneous shape and electron opacity. The nature of the vesicular particles and the significance of their presence in cilia and in the cytoplasm of some of the islet cells remain unknown. Among other possibilities, it is, however, suggested that the vesicular particles may represent secretory material.

Animals↗
PubMed1972

The metachronal wave of lateral cilia of Mytilus edulis.

The form of beat of cilia and the structure of the metachronal wave on the lateral gill epithelium of Mytulus edulis have been studied on living material by interference-contrast microscopy and stroboscopic illumination, and compared with the same features in rapid-fixed preparations studied by light microscopy and with the scanning electron microscope. The most striking finding is that the beat of the cilia is not planar, as previously assumed, but involves a sideways movement in the recovery stroke Previous reports on nonplanar ciliary beating from protozoan examples describe a planar effective stroke and a counterclockwise rotation in the recovery stroke; in this molluscan example there is a clockwise rotation in the recovery stroke The lateral inclination of the cilia in the recovery stroke is in the same direction as the propagation of the waves, and the orientation of cilia in the recovery stroke is thought to determine whether the waves move to the left or right of the direction of the effective stroke

Animals↗
PubMed1977

Electron microscope demonstration of tubulin in cilia and basal bodies of rat tracheal epithelium by the use of an antitubulin antibody.

It has been previously demonstrated that both cytoplasmic microtubules and the microtubules of cilia, flagella, and sperm tail contain tubulin. Although the morphology of cytoplasmic microtubules and that of axonemes differs in cells from which they have been isolated, the tubulin of the two structures shares physical and chemical properties. In some mammalian tissues, such as tracheal epithelium, cilia and basal bodies are difficult to isolate and characterize. The use of an enzyme-labeled immunoglobulin probe would facilitate identification and in situ localization of such proteins. Tubulin prepared from porcine brain by ion-exchange chromatography and from rat brain by the method of cyclic polymerization and depolymerization with subsequent disk gel electrophoresis with SDS were injected intravenously into rabbits. The animals were intermittently bled and the antisera extracted. The specificity of the antisera was proved by indirect immunofluorescence staining of the mitotic spindle, specific blocking of spindle staining by purified tubulin and not by other proteins, staining of 3T3 cytoplasmic microtubules, single line on immunoelectrophoresis, failure of control antisera to show any of these, and precipitation of antibody with all tubulin preparations and not with actin. We have shown by electron microscopy of ciliated cells of the tracheal epithelium stained with antitubulin by the indirect enzyme-labeled antibody method that the basal bodies, outer doublets, and central pair of the cilia contain tubulin. This indicates that tubulin in microtubules of cilia and basal bodies of rat tracheal epithelium is antigenically similar to tubulin extracted from cytoplasmic neurotubules of brains from the same species and from a different mammalian species. No other axonemal structures stained with the antitubulin. Three different preparations of tubulin from pigs and rats were used to immunize rabbits. All elicited similar antisera which gave identical staining patterns. The specificity of the staining was demonstrated by the absence of staining with immune serum absorbed with purified tubulin, the absence of staining with preimmune serum, and the absence of staining if any of the reagents were omitted during the staining reaction.

Animals↗
PubMed1981

Characterization of the cilia and ciliary membrane proteins of wild-type Paramecium tetraurelia and a pawn mutant.

Cilia and ciliary membranes were isolated from axenically grown, wild-type Paramecium tetraurelia strain 51s and from the extreme pawn mutant strain, d495, derived from this parental strain. Over 60 protein bands having molecular weights of 15 to greater than 300 kdaltons were detected by Coomassie Blue staining of whole cilia proteins separated by one-dimensional SDS polyacrylamide gel electrophoresis. About 30 of these protein bands were visible in Coomassie Blue-stained membrane separations. About 60 bands were detected by silver staining of one-dimensional gels of membrane proteins. Differences between Coomassie Blue-stained separations of wild-type and pawn mutant strain d495 membrane proteins were seen in the quantity of a band present at 43 kdaltons. Radioiodination of cell surface proteins labeled approximately 15 protein bands in both wild-type and mutant cilia. The major axonemal proteins were unlabeled. Six membrane glycoproteins were identified by staining one-dimensional separations with iodinated concanavalin A and lentil lectin, two lectins that specifically bind both glucose and mannose residues. Two major neutral sugar species present in an acid hydrolysate of the cilia preparation were tentatively identified as glucose and mannose by gas chromatography of the alditol acetate derivatives.

Animals↗
PubMed2003

Hypoxia regulates assembly of cilia in suppressors of Tetrahymena lacking an intraflagellar transport subunit gene.

We cloned a Tetrahymena thermophila gene, IFT52, encoding a homolog of the Chlamydomonas intraflagellar transport protein, IFT52. Disruption of IFT52 led to loss of cilia and incomplete cytokinesis, a phenotype indistinguishable from that of mutants lacking kinesin-II, a known ciliary assembly transporter. The cytokinesis failures seem to result from lack of cell movement rather than from direct involvement of ciliary assembly pathway components in cytokinesis. Spontaneous partial suppressors of the IFT52 null mutants occurred, which assembled cilia at high cell density and resorbed cilia at low cell density. The stimulating effect of high cell density on cilia formation is based on the creation of pericellular hypoxia. Thus, at least under certain conditions, ciliary assembly is affected by an extracellular signal and the Ift52p function may be integrated into signaling pathways that regulate ciliogenesis.

Algal Proteins↗
PubMed2006

The intraflagellar transport protein IFT20 is associated with the Golgi complex and is required for cilia assembly.

Eukaryotic cilia are assembled via intraflagellar transport (IFT) in which large protein particles are motored along ciliary microtubules. The IFT particles are composed of at least 17 polypeptides that are thought to contain binding sites for various cargos that need to be transported from their site of synthesis in the cell body to the site of assembly in the cilium. We show here that the IFT20 subunit of the particle is localized to the Golgi complex in addition to the basal body and cilia where all previous IFT particle proteins had been found. In living cells, fluorescently tagged IFT20 is highly dynamic and moves between the Golgi complex and the cilium as well as along ciliary microtubules. Strong knock down of IFT20 in mammalian cells blocks ciliary assembly but does not affect Golgi structure. Moderate knockdown does not block cilia assembly but reduces the amount of polycystin-2 that is localized to the cilia. This work suggests that IFT20 functions in the delivery of ciliary membrane proteins from the Golgi complex to the cilium.

Animals↗
PubMed2006

Caenorhabditis elegans DYF-2, an orthologue of human WDR19, is a component of the intraflagellar transport machinery in sensory cilia.

The intraflagellar transport (IFT) machinery required to build functional cilia consists of a multisubunit complex whose molecular composition, organization, and function are poorly understood. Here, we describe a novel tryptophan-aspartic acid (WD) repeat (WDR) containing IFT protein from Caenorhabditis elegans, DYF-2, that plays a critical role in maintaining the structural and functional integrity of the IFT machinery. We determined the identity of the dyf-2 gene by transgenic rescue of mutant phenotypes and by sequencing of mutant alleles. Loss of DYF-2 function selectively affects the assembly and motility of different IFT components and leads to defects in cilia structure and chemosensation in the nematode. Based on these observations, and the analysis of DYF-2 movement in a Bardet-Biedl syndrome mutant with partially disrupted IFT particles, we conclude that DYF-2 can associate with IFT particle complex B. At the same time, mutations in dyf-2 can interfere with the function of complex A components, suggesting an important role of this protein in the assembly of the IFT particle as a whole. Importantly, the mouse orthologue of DYF-2, WDR19, also localizes to cilia, pointing to an important evolutionarily conserved role for this WDR protein in cilia development and function.

Animals↗
PubMed1996

Effects of 3 MAC of halothane, enflurane and isoflurane on cilia beat frequency of human nasal epithelium in vitro.

We have measured the effects of three times the minimum alveolar concentration (MAC) of halothane, enflurane and isoflurane on cilia beat frequency of human nasal epithelial brushings from 18 healthy adult patients. Using the transmitted light technique and paired perfusion chambers, the cilia were exposed to 2.25% halothane, 5% enflurane or 3.6% isoflurane in air, or air alone, in a controlled and blinded manner. Over a 4-h observation period, cilia beat frequency of the samples exposed to inhalation anaesthetic agents demonstrated a significant reduction in frequency compared with controls exposed to air alone. Mean cilia beat frequency for the samples exposed to halothane was 9.3 (SEM 1.3) compared with its controls of 11.4 (1.0); for the samples exposed to enflurane, 10.9 (1.3) compared with its controls of 11.6 (1.2); and for the samples exposed to isoflurane, 10.8 (1.1) compared with its controls of 11.6 (1.2). There was a statistically significant difference between the samples exposed to all three volatile agents and their associated controls (halothane, P = 0.01; enflurane, P = 0.03; isoflurane, P = 0.01; nested repeated measures analysis of variance utilizing polynomial contrasts).

Adult↗
PubMed1997

Comparison of isoflurane with propofol on respiratory cilia.

We have investigated the effects of two techniques of clinical anaesthesia on human respiratory cilia by measuring cilia beat frequency of nasal tissue. In a randomized, controlled study, 13 patients undergoing either inhalation anaesthesia with isoflurane or total i.v. anaesthesia with propofol and alfentanil had nasal ciliated epithelial samples removed at the beginning and after 1 h of anaesthesia. Mean cilia beat frequency in the group anaesthetized with isoflurane changed significantly from 11.5 (95% confidence interval (CI) 10.7-12.2) Hz to 9.1 (8.1-10.1) Hz after anaesthesia whereas in the group anaesthetized with propofol and alfentanil there was a change from 11.5 (10.7-12.2) Hz to 11.0 (10.2-11.8) Hz (ns). The difference between the anaesthetic agents on cilia beat frequency was significant (MANOVA, P < 0.01). These data suggest that different anaesthetic agents may impair respiratory defence mechanisms to differing extents.

Adult↗
PubMed1988

A conditional mutant having paralyzed cilia and a block in cytokinesis is rescued by cytoplasmic exchange in Tetrahymena thermophila.

Nineteen mutants that are conditional for both the ability to regain motility following deciliation and the ability to grow were isolated. The mutations causing slow growth were placed into five complementation groups. None of the mutations appears to affect energy production as all mutants remained motile at the restrictive temperature. In three complementation groups protein synthesis and the levels of mRNA encoding alpha-tubulin or actin were largely unaffected at the restrictive temperature, consistent with the hypothesis that mutations in these three groups directly affect the assembly of functional cilia and growth. Complementation group 1 was chosen for further characterization. Both phenotypes were shown to be linked, suggesting they are caused by a single mutation. Group 1 mutants regenerated cilia at the restrictive temperature, but the cilia were nonmotile. This mutation also caused a block in cytokinesis at the restrictive temperature but did not affect nuclear divisions or DNA synthesis. The block in cell division was transiently rescued by wild-type cytoplasm exchanged when mutants were paired with wild-type cells during conjugation (round 1 of genomic exclusion). Thus, at least one mutation has been isolated that affects assembly of some microtubule-based structures in Tetrahymena (cilia during regeneration) but not others (nuclei divide at 38 degrees), and the product of this gene is likely to play a role in both ciliary function and in cytokinesis.

Actins↗
PubMed2002

Calcium-dependent phosphorylation of a protein in the frog olfactory cilia.

In the cilia of vertebrate olfactory sensory neurons, cytoplasmic Ca(2+) concentration increases in response to odorant stimulation, and this increase has been implicated to have important roles in the regulation of olfactory responses. Since protein phosphorylation is often a regulatory mechanism of biological reactions, we explored the effect of Ca(2+) on phosphorylation reactions in the frog olfactory cilia. First, we found that a 45-kDa phosphoprotein (p45) is predominantly phosphorylated in vitro in the isolated cilia in a Ca(2+)-dependent manner. However, later studies showed that the phosphorylation level of p45 is controlled by a dynamic equilibrium between phosphorylation and dephosphorylation. Although both activities are enhanced at high Ca(2+) concentrations (K(1/2) = approximately 2 microM in both reactions), the enhancement of dephosphorylation is relatively greater than that of phosphorylation. As a result, the steady phosphorylation level of p45 is lower at high than at low Ca(2+) concentration. The phosphorylation/dephosphorylation equilibrium was founed to involve protein kinases sensitive to zinc and heparin, and an unknown phosphatase(s). The present result suggests the presence of a novel Ca(2+)-signaling pathway that involves phosphorylation of p45 in the olfactory cilia.

Animals↗
PubMed2004

Loss of C. elegans BBS-7 and BBS-8 protein function results in cilia defects and compromised intraflagellar transport.

Bardet-Biedl syndrome (BBS) is a genetically heterogeneous developmental disorder whose molecular basis is largely unknown. Here, we show that mutations in the Caenorhabditis elegans bbs-7 and bbs-8 genes cause structural and functional defects in cilia. C. elegans BBS proteins localize predominantly at the base of cilia, and like proteins involved in intraflagellar transport (IFT), a process necessary for cilia biogenesis and maintenance, move bidirectionally along the ciliary axoneme. Importantly, we demonstrate that BBS-7 and BBS-8 are required for the normal localization/motility of the IFT proteins OSM-5/Polaris and CHE-11, and to a notably lesser extent, CHE-2. We propose that BBS proteins play important, selective roles in the assembly and/or function of IFT particle components. Our findings also suggest that some of the cardinal and secondary symptoms of BBS, such as obesity, diabetes, cardiomyopathy, and learning defects may result from cilia dysfunction.

Adaptor Proteins, Signal Transducing↗
PubMed2006

Hydrodynamic flow patterns and synchronization of beating cilia.

We calculate the hydrodynamic flow field generated far from a cilium which is attached to a surface and beats periodically. In the case of two beating cilia, hydrodynamic interactions can lead to synchronization of the cilia, which are nonlinear oscillators. We present a state diagram where synchronized states occur as a function of the distance of cilia and the relative orientation of their beat. Synchronized states occur with different relative phases. In addition, asynchronous solutions exist. Our work could be relevant for the synchronized motion of cilia generating hydrodynamic flows on the surface of cells.

Animals↗
PubMedDate not supplied

Mosaicism of dynein in spermatozoa and cilia and fibrous sheath aberrations in an infertile man.

Male patients displaying an immotile or almost immotile sperm population are the object of an interdisciplinary study concerning a ciliary mutant that induces the "Immotile-Cilia Syndrome". Development and function of both sperm flagella and cilia are normally affected because of disturbances of the 9 + 2-arrangement. During this program, clinical, physiological, genetical and ultrastructural investigations were done. The ultrastructure of immotile spermatozoa of an infertile man did not reveal inner and outer dynein arms. Lack of the ATPase dynein which is essential for movement of the 9 + 2-axoneme, is typical for the above syndrome. In addition, symmetry of the fibrous sheath of the spermatozoa was very abnormal. The pneumologist examined normal lung function, where the ultrastructure of the cilia of the nasal mucosa displayed the dynein arms. Analysis of family tree and chromosomes by the geneticist also gave a normal result. As revealed by this infertile patient it seems likely that expression of dynein must not be identical in both germ cells and somatic cells. Such variations are therefore regarded as additional forms of the "Immotile-Cilia syndrome". Asymmetric fibrous sheaths are thought to be a result of immotile spermatid flagella, leading to an abnormal arrangement of the accessory axonemal structures. Normal early spermatid flagella of man and rat show specific movements.

Adenosine Triphosphatases↗
PubMed1974

The distribution, activity, and function of the cilia in the frog brain.

1. The distribution, activity, and function of the cilia in the brain was studied using in vitro preparations of the frog choroid plexuses and ependyma.2. Scanning electron microscopy revealed that twenty to forty cilia, about 20 mum long, project from the cells of the choroidal epithelium and ependyma into the ventricular system of the brain.3. These cilia beat at a constant frequency which ranged from 5 to 20 c/s. Ciliary activity was enhanced by ATP, cyclic AMP, theophylline, and acetylcholine, and was depressed by DNP, IAA, Ni(2+), La(3+), and Co(2+).4. Ciliary motion produced a flow of c.s.f. over the surface of the cells lining the ventricles, and in the choroid plexus this flow reduced the effective thickness of the unstirred layer adjacent to the epithelium by about 100 mum.5. These results are discussed in relation to the factors that control the frequency of ciliary beating, and the role of the cilia in the circulation of the c.s.f.

Acetylcholine↗
PubMed1981

Ultrastructural abnormalities of bronchial cilia in children with recurrent airway infections and bronchiectasis.

Anomalies of the bronchial cilia were studied in 5 children with recurrent pulmonary infections. Case 1 had Kartagener's syndrome and an absence of the inner and outer dynein arms in most cilia, although a few shortened and even some normal arms could be seen. Cases 2 and 3 had unilateral bronchiectasis without family history of Kartagener's syndrome. Serial studies of the bronchial epithelium at times showed a bilateral lack of the inner dynein arms and a partial lack of outer arms. These abnormalities persisted in these 2 children after they had recovered from the acute pulmonary infection but disappeared after 6-8 months of antibiotic treatment. Cases 4 and 5 had recurrent pulmonary infections without bronchiectasis and many shortened outer dynein arms could be seen, but these anomalies disappeared after recovery. In all 5 children such architectural ciliary anomalies were present as megacilia, fused cilia, naked cilia, and completely disorganised axonemas. These architectural defects were particularly numerous in the children without bronchiectasis. Our observations suggest that anomalies of the bronchial ciliary microtubular system may not only be congenital but may also be acquired; this might well help to explain some cases of repeated respiratory tract infection and bronchiectasis.

Adolescent↗
PubMed2007

Overview of structure and function of mammalian cilia.

Cilia are membrane-bounded, centriole-derived projections from the cell surface that contain a microtubule cytoskeleton, the ciliary axoneme, surrounded by a ciliary membrane. Axonemes in multiciliated cells of mammalian epithelia are 9 + 2, possess dynein arms, and are motile. In contrast, single nonmotile 9 + 0 primary cilia are found on epithelial cells, such as those of the kidney tubule, but also on nonepithelial cells, such as chondrocytes, fibroblasts, and neurons. The ciliary membranes of all cilia contain specific receptors and ion channel proteins that initiate signaling pathways controlling motility and/or linking mechanical or chemical stimuli, including sonic hedgehog and growth factors, to intracellular transduction cascades regulating differentiation, migration, and cell growth during development and in adulthood. Unique motile 9 + 0 cilia, found during development at the embryonic node, determine left-right asymmetry of the body.

Animals↗
PubMed1995

Presence and possible implications of open-ring centrioles, multiple basal centrioles and basal cilia in neonatal hamster bronchioles.

In studying epithelio-stromal interactions in the neonatal hamster lung, basal centrioles, basal cilia and open-ring centrioles attracted our attention. The same structures are also found in bronchiolar epithelium of a 3-week-old fawn-hooded rat, indicating that they are not specific phenomena of hamster lung. Although cells with basal centrioles are not common, they are more readily observed in a sample of bronchioles at the second postnatal week as compared to the fourth neonatal day. The centrioles may often possess two satellites and function as microtubule organizing centers (MTOC). Open-ring centrioles are mature structures and also serve as MTOC. Apparently, triplets may de novo be added to and/or degraded from an open-ring centriole. The basal cilia and the cilia in the ciliated vacuoles/cysts, have a normal axonemal structure and may have a common origin from the basal centrioles. It is, however, unclear whether or not they are primary-like cilia. In addition, presence of the multiple basal MTOC and some morphologic features point to a high microtubule-associated vesicular transport activity and probably indicate an elevated interaction with the interstitium; thus, the cells have a different polarization then normal ciliated cells.

Animals↗