共查询到20条相似文献,搜索用时 15 毫秒
1.
Cilia-lacking respiratory cells in ciliary aplasia 总被引:1,自引:0,他引:1
M M de Santi C Gardi G Barlocco M Canciani G Mastella G Lungarella 《Biology of the cell / under the auspices of the European Cell Biology Organization》1988,64(1):67-70
This report describes the ultrastructural alterations observed in the nasal and bronchial mucosa of an 11-yr-old male suffering from immotile cilia syndrome (ICS). The morphological features observed in this patient are consistent with a ciliary aplasia. In fact, ciliated cells appeared to be replaced by columnar cells lacking cilia and basal bodies, and bearing on their surface cilium-like projections without any internal axonemal structure. In spite of the absence of basal bodies, centrioles, and kinocilia, these cells unexpectedly showed mature striated roots and centriolar precursor material scattered throughout the apical cytoplasm. These data suggest that control over basal body assembly is distinct from control over striated root formation. The presence of the above-reported structures in cells otherwise presenting many morphological features of normal ciliated cells is discussed on the basis of current knowledge of respiratory cilia biogenesis. 相似文献
2.
N A Shanina V I Gel'fand I I Dykhanov N G Bronza N N Rozinova 《Biulleten' eksperimental'no? biologii i meditsiny》1986,101(1):84-86
The method of extraction of ciliated epithelium from biopsy samples of human bronchial mucosa with glycerol is suggested. Permeabilized cilia of glycerol-extracted cells can be easily reactivated by exogenous ATP. This method was used for the study of ciliary dyskinesia in patients with chronic lung diseases. It was shown that in patients with Kartagener's syndrome neither freshly-isolated, nor glycerol-extracted ATP-treated cilia were motile. On the other hand, in some patients with bronchial asthma ATP reactivated glycerol-extracted cilia, while cilia of freshly-isolated cells remained immotile. The study shows that glycerol permeabilization and reactivation by ATP can be used for the analysis of cilial contractile apparatus in patients with chronic lung disease. 相似文献
3.
4.
Khristofor Agassandian Milan Patel Marianna Agassandian Karina E. Steren Kamal Rahmouni Val C. Sheffield J. Patrick Card 《PloS one》2014,9(4)
Bardet-Biedl syndrome (BBS) is a genetically heterogeneous inherited human disorder displaying a pleotropic phenotype. Many of the symptoms characterized in the human disease have been reproduced in animal models carrying deletions or knock-in mutations of genes causal for the disorder. Thinning of the cerebral cortex, enlargement of the lateral and third ventricles, and structural changes in cilia are among the pathologies documented in these animal models. Ciliopathy is of particular interest in light of recent studies that have implicated primary neuronal cilia (PNC) in neuronal signal transduction. In the present investigation, we tested the hypothesis that areas of the brain responsible for learning and memory formation would differentially exhibit PNC abnormalities in animals carrying a deletion of the Bbs4 gene (Bbs4-/-). Immunohistochemical localization of adenylyl cyclase-III (ACIII), a marker restricted to PNC, revealed dramatic alterations in PNC morphology and a statistically significant reduction in number of immunopositive cilia in the hippocampus and amygdala of Bbs4-/- mice compared to wild type (WT) littermates. Western blot analysis confirmed the decrease of ACIII levels in the hippocampus and amygdala of Bbs4-/- mice, and electron microscopy demonstrated pathological alterations of PNC in the hippocampus and amygdala. Importantly, no neuronal loss was found within the subregions of amygdala and hippocampus sampled in Bbs4-/- mice and there were no statistically significant alterations of ACIII immunopositive cilia in other areas of the brain not known to contribute to the BBS phenotype. Considered with data documenting a role of cilia in signal transduction these findings support the conclusion that alterations in cilia structure or neurochemical phenotypes may contribute to the cognitive deficits observed in the Bbs4-/- mouse mode. 相似文献
5.
A Németh 《Acta morphologica Academiae Scientiarum Hungaricae》1976,24(3):247-259
In hydrophilic media of high refraction index the cilia of the epithelium of the tracheo-bronchial mucous membrane and the mucous film demonstrate birefringence positive to the longitudinal axis of the mucous membrane owing to the identical orientation the glycoprotein macromolecules and of the fatty acid chains of the ciliary membrane. By means of toluidine blue staining the mucous film and the ciliary zone can be studied selectively by polarization microscopy. The opposite optical character suggests that the dye molecules are perpendicularly connected to the long glycoprotein chains of the mucous, and there is an oriented connection perpendicular to the longitudinal axis of the cilium in its lipid membrane. In smears of bronchial secretion or mucous membrane scrapings after staining, with toluidine blue pH 7.0, the selective optical reaction of the cilia provides a possibility for the study of isolated desquamated ciliary cells. The reaction is given by the lipid membrane boundary of the cilia. In embedded, lipid-extracted sections, the cilia and the mucous film show a minimum of birefringence in the unstained state. After toluidine blue staining or aldehyde-bisulphite-toluidine blue (ABT) reaction the mucoid surface and the ciliary zone display an opposite optical reaction originating from the dehydrated mucoproteins adsorbed onto the surface of lipid-extracted cilia. 相似文献
6.
It has been shown that bronchial mucus from patients with bronchial asthma and chronic bronchitis can produce a ciliostatic effect when incubated with strips of frog palate mucosa. This effect can more often be found during clinical exacerbation and is supposed to be reversible. The ciliostatically active samples of bronchial mucus taken from asthmatic patients markedly inhibit reactivation glycerol models of frog's ciliated epithelium. It can therefore be suggested that the found activity acts directly on tubulin-dynein complex of the cilia. In contrast to these findings, the ciliostatically active samples of bronchial of glycerol models and therefore possibly act through some other mechanism. 相似文献
7.
Petra M. de Jong Marianne A. J. A. van Sterkenburg Johanna A. Kempenaar Joop H. Dijkman Maria Ponec 《In vitro cellular & developmental biology. Animal》1993,29(5):379-387
Summary In the present study we describe the establishment of serial cultures of human bronchial epithelial cells derived from biopsies
obtained by fiberoptic bronchoscopy. The cell cultures were initiated from small amounts of material (2 mm forceps biopsies)
using either explants or epithelial cell suspensions in combination with a feeder-layer technique. The rate of cell proliferation
and the number of passages (up to 8 passages) achieved were similar, irrespective of whether the explants or dissociated cells
were used. To modulate the extent of differentiation, the bronchial epithelial cells were cultured either under submerged,
low calcium (0.06 mM) (proliferating), normal calcium (1.6 mM) (differentiation enhancing) conditions, or at the air-liquid interface. Characterization of the bronchial epithelial cell
cultures was assessed on the basis of cell morphology, cytokeratin expression, and ciliary activity. The cells cultured under
submerged conditions formed a multilayer consisting of maximally three layers of polygonal-shaped, small cuboidal cells, an
appearance resembling the basal cells in vivo. In the air-exposed cultures, the formed multilayer consisted of three to six
layers exhibiting squamous metaplasia. The cytokeratin profile in cultured bronchial epithelial cells was similar in submerged
and air-exposed cultures and comparable with the profile found in vivo. In addition to cytokeratins, vimentin was co-expressed
in a fraction of the subcultured cells. The ciliary activity was observed in primary culture, irrespective of whether the
culture had been established from explants or from dissociated cells. This activity was lost upon subculturing and it was
not regained by prolongation of the culture period. In contrast to submerged cultures and despite the squamous metaplasia
appearance, the cells showed a reappearance of cilia when cultured at the air-liquid interface. Human bronchial epithelial
cell cultures can be a representative model for controlling the mechanisms of regulation of bronchial epithelial cell function. 相似文献
8.
C F Clary-Meinesz J Cosson P Huitorel B Blaive 《Biology of the cell / under the auspices of the European Cell Biology Organization》1992,76(3):335-338
Even though all human respiratory cilia are similar in structure, they experience a wide range of temperatures between the initial part of the nasal fossae which behave as heat exchangers and the inferior part of the trachea, particularly when we inhale exceedingly cold or hot air. The ciliary beat frequency of ciliated cells from human nasal mucosa and from bronchial mucosa averages 8 Hz when measured at room temperature. In the present study we compared the ciliary beat frequency of human cells from nasal and tracheal mucosa brushings at different temperatures from 5 degrees C to 50 degrees C using two different techniques, ex vivo and in vitro: ex vivo in culture medium less than 24 h after sampling and in vitro after demembranation and reactivation according to a standard procedure developed in our laboratory. Measuring the ATP-reactivated ciliary beat frequency allowed us to check the thermal parameters of the dynein ATPase and all the axonemal machinery. No significant difference in frequency was observed between nasal fossae cilia and tracheal cilia when comparing extreme temperatures in both experimental procedures. 相似文献
9.
Seiichi Tanida Toru Hasegawa Eiji Higashide 《Bioscience, biotechnology, and biochemistry》2013,77(8):1847-1853
Cilia regeneration with deciliated Tetrahymena pyriformis W was tested to determine the antitubulinic activities of ansamitocins and related compounds in a microbial system. Various factors interfered with the regeneration process, i.e. excess shearing force in deciliation procedure, high temperature (32°C or above), high or low pH (pH 9 or 5), and exogenous divalent cations, such as Zn2+, Mn2+, Cu2+ and Co2+. Under suitable conditions, cilia regeneration was completed in about 60 min of incubation, and a number of newly formed cilia were observed around the cell surface. When ansamitocins were added to the recovery solution, cilia regeneration was completely suppressed without alterations in cell shape or the surface structure of deciliated Tetrahymena. In this assay system, the inhibitory activity of ansamitocins was slightly stronger than that of maytansine. 相似文献
10.
Takahashi M Sano T Yamaoka K Kamimura T Umemoto N Nishitani H Yasuoka S 《Histochemistry and cell biology》2001,115(3):181-187
Human airway trypsin-like protease (HAT) has been isolated from mucoid sputum of patients with chronic airway diseases. In order to clarify the cellular source of this novel protease in the human airway, we examined the localization of immunoreactive HAT in bronchial tissues obtained at surgery and fixed in 4% paraformaldehyde using an extremely sensitive immunohistochemical technique called a catalyzed signal amplification method and a monoclonal antibody against recombinant HAT. HAT immunoreactivity was demonstrated in cytoplasm of ciliated cells of bronchial epithelium and/or at the basal part of cilia. No positive reaction was found in submucosal glands or mast cells. The heterogeneous distribution of HAT immunoreactivity within the bronchial epithelium indicates that its expression might be changeable and that it might be closely related to the physiological status of the airway epithelium. Non-specific but intense reaction caused by endogenous avidin-binding activity (EABA) was selectively detected in submucosal glands, but was effectively blocked by successive treatments with avidin and biotin. These results indicate that HAT may be synthesized in the ciliated cells and that it may play some physiological roles within the epithelial layer and on the airway surface. It is necessary to keep in mind that some cells show strong EABA, especially when a highly sensitive immunohistochemical technique is applied. 相似文献
11.
Justina Hessel Jonna Heldrich Jennifer Fuller Michelle R. Staudt Sharon Radisch Charleen Hollmann Ben-Gary Harvey Robert J. Kaner Jacqueline Salit Jenny Yee-Levin Sriram Sridhar Sreekumar Pillai Holly Hilton Gerhard Wolff Hans Bitter Sudha Visvanathan Jay Fine Christopher S. Stevenson Ronald G. Crystal Ann E. Tilley 《PloS one》2014,9(1)
Smoking and COPD are associated with decreased mucociliary clearance, and healthy smokers have shorter cilia in the large airway than nonsmokers. We hypothesized that changes in cilia length are consistent throughout the airway, and we further hypothesized that smokers with COPD have shorter cilia than healthy smokers. Because intraflagellar transport (IFT) is the process by which cilia of normal length are produced and maintained, and alterations in IFT lead to short cilia in model organisms, we also hypothesized that smoking induces changes in the expression of IFT-related genes in the airway epithelium of smokers and smokers with COPD. To assess these hypotheses, airway epithelium was obtained via bronchoscopic brushing. Cilia length was assessed by measuring 100 cilia (10 cilia on each of 10 cells) per subject and Affymetrix microarrays were used to evaluate IFT gene expression in nonsmokers and healthy smokers in 2 independent data sets from large and small airway as well as in COPD smokers in a data set from the small airway. In the large and small airway epithelium, cilia were significantly shorter in healthy smokers than nonsmokers, and significantly shorter in COPD smokers than in both healthy smokers and nonsmokers. The gene expression data confirmed that a set of 8 IFT genes were down-regulated in smokers in both data sets; however, no differences were seen in COPD smokers compared to healthy smokers. These results support the concept that loss of cilia length contributes to defective mucociliary clearance in COPD, and that smoking-induced changes in expression of IFT genes may be one mechanism of abnormally short cilia in smokers. Strategies to normalize cilia length may be an important avenue for novel COPD therapies. 相似文献
12.
E Boisvieux-Ulrich M C Lainé D Sandoz 《Biology of the cell / under the auspices of the European Cell Biology Organization》1989,67(1):67-79
Oviduct implants from quails which were primarily stimulated in vivo by estrogen so as to induce ciliogenesis in some epithelial cells were cultured in vitro in the presence or absence of colchicine or nocodazole. After 24 or 48 hr of culture, implants were examined by transmission and scanning electron microscopy to determine drug-induced alterations in ciliogenesis. After 24 hr of 10(-5) M colchicine treatment, the formation of basal bodies was totally inhibited, though the precursor material of generative complexes was unchanged. The inhibitory effect was not reversed when colchicine was removed in a 24 hr recovery culture. Treatment with 10(-6) M nocodazole for 24 hr, partially inhibited the assembly of basal bodies, which exhibited altered morphology. The assembly of basal bodies was restored during the 24 hr recovery period, after removal of nocodazole. Colchicine and nocodazole did not prevent polarized migration towards the apical surface of basal bodies formed prior to drug treatment. They anchored to the plasma membrane, but the formation of cilia was strongly disturbed in the presence of the drug. Numerous cells possessed anchored basal bodies which failed to induce the formation of cilia. The elongation of cilia was inhibited, as seen by their abnormal capping structure. In the enlarged tip, microtubules diverged. In contrast, these very short cilia possessed a mature ciliary necklace which was constructed during drug treatment. Differentiation of this membrane ciliary structure appeared to be unrelated to axoneme growth. 相似文献
13.
Kerry A. Miller Casey J. Ah-Cann Megan F. Welfare Tiong Y. Tan Kate Pope Georgina Caruana Mary-Louise Freckmann Ravi Savarirayan John F. Bertram Michael S. Dobbie John F. Bateman Peter G. Farlie 《PLoS genetics》2013,9(8)
Cilia are architecturally complex organelles that protrude from the cell membrane and have signalling, sensory and motility functions that are central to normal tissue development and homeostasis. There are two broad categories of cilia; motile and non-motile, or primary, cilia. The central role of primary cilia in health and disease has become prominent in the past decade with the recognition of a number of human syndromes that result from defects in the formation or function of primary cilia. This rapidly growing class of conditions, now known as ciliopathies, impact the development of a diverse range of tissues including the neural axis, craniofacial structures, skeleton, kidneys, eyes and lungs. The broad impact of cilia dysfunction on development reflects the pivotal position of the primary cilia within a signalling nexus involving a growing number of growth factor systems including Hedgehog, Pdgf, Fgf, Hippo, Notch and both canonical Wnt and planar cell polarity. We have identified a novel ENU mutant allele of Ift140, which causes a mid-gestation embryonic lethal phenotype in homozygous mutant mice. Mutant embryos exhibit a range of phenotypes including exencephaly and spina bifida, craniofacial dysmorphism, digit anomalies, cardiac anomalies and somite patterning defects. A number of these phenotypes can be attributed to alterations in Hedgehog signalling, although additional signalling systems are also likely to be involved. We also report the identification of a homozygous recessive mutation in IFT140 in a Jeune syndrome patient. This ENU-induced Jeune syndrome model will be useful in delineating the origins of dysmorphology in human ciliopathies. 相似文献
14.
《Tissue & cell》2016,48(6):588-595
In the present study, we examined the morphology of cilia and expression of the dynein intermediate chain 2 (DNAI2) in the oviduct of non-obese diabetic (NOD) mice. Results obtained with immunohistochemistry showed that DNAI2 expression was reduced in oviducts of diabetic NOD (dNOD) mice, as compared to that observed in the normoglycemic NOD (cNOD) group, especially in the acyclic dNOD mice. Oviductal cilia of dNOD mice appeared to be reduced in number. Results obtained with Western blot analysis revealed that the expression of DNAI2 protein was significantly less in oviducts of dNOD mice as compared to that of cNOD mice corroborating the results obtained with immunohistochemistry. Electron microscopic examination and quantitative imaging of thin sections of Epon-embedded oviducts of both dNOD and cNOD mice confirmed the reduction of the number of cilia in the oviduct of the dNOD group which also displayed aberrant axonemal ultrastructure, including disorganization of the axoneme and alteration of microtubule doublets into singlets as well as disruption of the plasma membrane in many cilia. Taken together, the present findings suggest that structural alterations of oviductal cilia in female diabetic NOD mice might be detrimental to the normal function of these particular cell structures in gamete transport. 相似文献
15.
Reduced primary cilia length and altered Arl13b expression are associated with deregulated chondrocyte Hedgehog signaling in alkaptonuria
下载免费PDF全文
![点击此处可从《Journal of cellular physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Stephen D. Thorpe Silvia Gambassi Clare L. Thompson Charmilie Chandrakumar Annalisa Santucci Martin M. Knight 《Journal of cellular physiology》2017,232(9):2407-2417
Alkaptonuria (AKU) is a rare inherited disease resulting from a deficiency of the enzyme homogentisate 1,2‐dioxygenase which leads to the accumulation of homogentisic acid (HGA). AKU is characterized by severe cartilage degeneration, similar to that observed in osteoarthritis. Previous studies suggest that AKU is associated with alterations in cytoskeletal organization which could modulate primary cilia structure/function. This study investigated whether AKU is associated with changes in chondrocyte primary cilia and associated Hedgehog signaling which mediates cartilage degradation in osteoarthritis. Human articular chondrocytes were obtained from healthy and AKU donors. Additionally, healthy chondrocytes were treated with HGA to replicate AKU pathology (+HGA). Diseased cells exhibited shorter cilia with length reductions of 36% and 16% in AKU and +HGA chondrocytes respectively, when compared to healthy controls. Both AKU and +HGA chondrocytes demonstrated disruption of the usual cilia length regulation by actin contractility. Furthermore, the proportion of cilia with axoneme breaks and bulbous tips was increased in AKU chondrocytes consistent with defective regulation of ciliary trafficking. Distribution of the Hedgehog‐related protein Arl13b along the ciliary axoneme was altered such that its localization was increased at the distal tip in AKU and +HGA chondrocytes. These changes in cilia structure/trafficking in AKU and +HGA chondrocytes were associated with a complete inability to activate Hedgehog signaling in response to exogenous ligand. Thus, we suggest that altered responsiveness to Hedgehog, as a consequence of cilia dysfunction, may be a contributing factor in the development of arthropathy highlighting the cilium as a novel target in AKU. 相似文献
16.
H. -D. Rott 《Human genetics》1979,46(3):249-261
Summary Kartagener's syndrome (KS) is a hereditary disease with typical symptoms of situs inversus, bronchiectasis, and chronic infections of the nasal mucosa. Autosomal recessive inheritance cannot be doubted on account of repeated observations of affected sibs and parental cansanguinity. The bronchopulmonary symptoms in sibs, however, cannot be explained by this mode of inheritance.Recent clinical findings and electron microscope investigations suggest that KS is a special form of manifestation within the immotile cilia syndrome. This disease combines the typical bronchial and nasal symptoms of KS with sterility in the male due to immotile sperm tails and, as a facultative symptom, situs inversus. Thus, sibs with bronchiectasis but without situs inversus are also classified under this syndrome. The symptoms mentioned are caused by an abnormal morphology of bronchial cilia and sperm tails, which can be demonstrated by electron microscopy. The dynein arms normally attached to the nine microtubular doublets and providing a normal ciliary movement are lacking.It is assumed that during early embryonic life ciliary beats in the growing embryo determine the type of laterality. When ciliary movements are absent laterality may develop fortuitously, thus effecting a situs inversus in about half the affected cases. The numerical evaluation of pedigrees from the literature supports this assumption. 相似文献
17.
Long W. M.; Sprung C. L.; el Fawal H.; Yerger L. D.; Eyre P.; Abraham W. M.; Wanner A. 《Journal of applied physiology》1985,59(1):254-261
The effects of aerosolized 5% histamine (10 breaths) on bronchial artery blood flow (Qbr), airflow resistance (RL), and pulmonary and systemic hemodynamics were studied in mechanically ventilated sheep anesthetized with pentobarbital sodium. Histamine increased mean Qbr and RL to 252 +/- 45 and 337 +/- 53% of base line, respectively. This effect was significantly different from base line for 30 min after challenge. The histamine-induced increase in RL was blocked by pretreatment with the histamine H1 receptor antagonist, chlorpheniramine, whereas the histamine-induced elevation in Qbr was prevented by the H2 antagonist, metiamide. Both responses were blocked only when both antagonists were present. Changes in Qbr were not directly associated with alterations in systemic and pulmonary hemodynamics or arterial blood gas composition. In vitro histamine caused a dose-dependent contraction of ovine bronchial artery strips that was prevented by H1 antagonist. The H2 agonist, impromidine, caused relaxation of precontracted arterial strips and was more potent and efficacious than histamine, whereas H1 agonists failed to elicit a relaxant response. Thus these findings indicate that histamine aerosol induces a vasodilation in the bronchial vascular bed; histamine has a direct effect on Qbr that is independent of alterations in RL, systemic and pulmonary hemodynamics, or arterial blood gas composition; and, histamine-induced bronchoconstriction is mediated predominantly by H1-receptors, whereas increased Qbr is controlled predominantly by H2-receptors, probably located in resistance vessels. This local effect of histamine on Qbr may have important implications in the pathophysiology of bronchial asthma and pulmonary edema. 相似文献
18.
J. COSSON 《Cell biology international》1996,20(2):83-94
Beating of cilia and flagellae allows movement of the fluid surrounding isolated cells (for example: protists) or epithelia (bronchial tissue) but is also responsible for the movement of unicellular organisms in this medium (such as spermatozoa or protists). This paper aims to describe: (1) the biochemical and structural elements of the ‘9 +2’ structure called the axoneme; (2) the mechanisms of wave generation and propagation along the axoneme of cilia and flagellae are then described, stating that in most models of wave propagation, a clear distinction is made between the dynein-dependent microtubule sliding which represents the oscillatory motor and the bending mechanism which regulates wave propagation. In current models, the bending propagation is supported by a bind /relax cyclic mechanism which propagates in register, but frame-shifted, with the powering action of the dynein motor along the axoneme. While a large amount of knowledge was accumulated about the motor, little is known about the resisting elements regulating the bending. (3) The present study also puts forward ideas as to how these organelles have been highly conserved throughout eucaryotic evolution, and concludes with suggestions for further fields of investigation into this unique mechanical device used for cell movement. 相似文献
19.
Banizs B Pike MM Millican CL Ferguson WB Komlosi P Sheetz J Bell PD Schwiebert EM Yoder BK 《Development (Cambridge, England)》2005,132(23):5329-5339
Cilia are complex organelles involved in sensory perception and fluid or cell movement. They are constructed through a highly conserved process called intraflagellar transport (IFT). Mutations in IFT genes, such as Tg737, result in severe developmental defects and disease. In the case of the Tg737orpk mutants, these pathological alterations include cystic kidney disease, biliary and pancreatic duct abnormalities, skeletal patterning defects, and hydrocephalus. Here, we explore the connection between cilia dysfunction and the development of hydrocephalus by using the Tg737orpk mutants. Our analysis indicates that cilia on cells of the brain ventricles of Tg737orpk mutant mice are severely malformed. On the ependymal cells, these defects lead to disorganized beating and impaired cerebrospinal fluid (CSF) movement. However, the loss of the cilia beat and CSF flow is not the initiating factor, as the pathology is present prior to the development of motile cilia on these cells and CSF flow is not impaired at early stages of the disease. Rather, our results suggest that loss of cilia leads to altered function of the choroid plexus epithelium, as evidenced by elevated intracellular cAMP levels and increased chloride concentration in the CSF. These data suggest that cilia function is necessary for regulating ion transport and CSF production, as well as for CSF flow through the ventricles. 相似文献
20.
Wyatt TA Forgèt MA Adams JM Sisson JH 《American journal of physiology. Lung cellular and molecular physiology》2005,288(3):L546-L551
Previously, we have shown that the ATPase-dependent motion of cilia in bovine bronchial epithelial cells (BBEC) can be regulated through the cyclic nucleotides, cAMP via the cAMP-dependent protein kinase (PKA) and cGMP via the cGMP-dependent protein kinase (PKG). Both cyclic nucleotides cause an increase in cilia beat frequency (CBF). We hypothesized that cAMP and cGMP may act directly at the level of the ciliary axoneme in BBEC. To examine this, we employed a novel cell-free system utilizing detergent-extracted axonemes. Axoneme movement was whole-field analyzed digitally with the Sisson-Ammons video analysis system. A suspension of extracted axonemes remains motionless until the addition of 1 mM ATP that establishes a baseline CBF similar to that seen when analyzing intact ciliated BBEC. Adding 10 microM cAMP or 10 microM cGMP increases CBF beyond the established ATP baseline. However, the cyclic nucleotides did not stimulate CBF in the absence of ATP. Therefore, the combination of cAMP and cGMP augments ATP-driven CBF increases at the level of isolated axoneme. 相似文献