Light and electron microscopic observations on ciliated vacuoles and cysts in the oviductal and endocervical epithelia of the rabbit

Ciliated vacuoles and intraepithelial cysts have been observed in oviductal and endocervical epithelia of rabbits. In this study, rabbits under various hormonal conditions were studied by light and transmission electron microscopy and tissue culture in an attempt to determine their distribution and origin. Ciliated vacuoles most frequently lay in the basal cytoplasm, below or beside the nucleus, and very close to the basal lamina. A few were apically located. Their average diameter was 8.8 by 5.1 microns. Cilia and microvilli projected into the vacuolar lumen. These vacuoles were located intracellularly as evidenced first by the degeneration of both their cilia and microvilli and the moderately dense matrix that often filled the vacuolar lumen, as observed by electron microscopy. Secondly, phase microscopy of the living endocervical epithelium allowed us to observe the beating of the cilia within the vacuoles, not on the surface of such cells. Thirdly, ruthenium red stained the surface glycocalyx of ciliated and secretory cells, but not that of the cilia and microvilli within the vacuoles. The intraepithelial cysts were not observed in all tissue blocks. The largest numbers were found in ovariectomized animals treated for 3 and 5 days with estradiol. More were seen in the isthmus and cervix than in the fimbria and ampulla. The cysts were located most often within the epithelium along the sides of, and at the bases of, the mucosal folds. They were lined by flattened epithelium of various combinations of secretory and ciliated cells. An unusual cell type was associated with some of the cysts and ciliated vacuoles. Its cytoplasm contained aggregates of mitochondria and vesicles whose contents varied in density. Although the genesis of the ciliated vacuoles is not certain, our results indicate that they may arise from aberrant positioning of proliferating procentrioles or from a defect in targeting or transporting the centrioles to the apical plasma membrane to serve as basal bodies. Fusion of adjacent ciliated vacuoles with lumina lined by secretory cells having deep apical invaginations appeared to contribute to the formation of cysts.     

Cilia-lacking respiratory cells in ciliary aplasia

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.     

Organization of actin microfilaments in the apical border of oviduct ciliated cells

Actin microfilaments were localized in quail oviduct ciliated cells using decoration with myosin subfragment S1 and immunogold labeling. These polarized epithelial cells show a well developed cytoskeleton due to the presence of numerous cilia and microvilli at their apical pole. Most S1-decorated microfilaments extend from the microvilli downward towards the upper part of the ciliary striated rootlets with which they are connected. From the microvillous roots, a few microfilaments connect the proximal part of the basal body or the basal foot associated with the basal body. Microfilament polarity is shown by S1 arrowheads pointing away from the microvillous tip to the cell body. Furthermore, short microfilaments are attached to the plasma membrane at the anchoring sites of basal bodies and run along the basal body. The polarity of these short microfilaments is directed from the basal body anchoring fibers downward to the cytoplasm. At the cell periphery, microfilaments from microvillous roots and ciliary apparatus are connected with those of the circumferential actin belt which is associated with the apical zonula adhaerens. Together with the other cytoskeletal elements, the microfilaments increase ciliary anchorage and could be involved in the coordination of ciliary beating. Moreover, microvilli surrounding the cilia probably modify ciliary beating by offering resistance to cilium bending. The presence of microvilli could explain the fact that mainly the upper part of the cilia appanars to be involved in the axonemal bending in metazoan ciliated cells.     

Observations on the solitary cilium of rabbit oviductal epithelium: its motility and ultrastructure

Solitary cilia have been observed on rabbit oviductal epithelial cells. In tissue cultures of fimbrial epithelium of 3- and 4-day-old animals observed by phase microscopy, most of these single cilia exhibited a vortical or funnel-type movement while others had the usual to-and-fro motility. Primary cilia are usually considered immotile. Transmission electron microscopy of specifically identified single cilia revealed differences between the ciliary shafts and basal bodies of the single cilia as compared to those of mature oviductal ciliated cells. The basal body of the solitary cilium often had at least two triangular, striated, basal foot processes, lacked electron-dense satellite material around its basal end, and occasionally had striated rootlets. In contrast, the cilia of mature ciliated cells had only one basal foot, exhibited much electron-dense satellite material, and lacked rootlets. Cross sections of the single cilia showed patterns of microtubules different from the usual 9 + 2 axonemal complexes of normal cilia and included 9 + 0, 10 + 2 singlets, 7 + 2 doublets, and 8 + 1 doublet and 2 singlets; one did have the usual 9 + 2 arrangement. We postulate that the presence of more than one basal foot process may be responsible for the vortical motility observed. The primary cilia are shorter than normal cilia; the longest one measured was 1.86 micron in length, 0.28 micron in width at its base, and 0.14 micron at its tip. Based on the light-microscopic, scanning-electron-microscopic and transmission-electron-microscopic observations, such solitary cilia were observed more frequently in the oviductal tissues of the 3- to 4-day postnatal rabbits grown in tissue culture and in ovariectomized and ovariectomized/progesterone-treated adult animals than in estrous, ovulatory, or ovariectomized/estradiol-treated rabbits.     

An ultrastructural study of primary cilia,abnormal cilia and ciliary knobs from the ciliated cells of the guinea-pig trachea

Summary Single primary cilia are found in developing as well as mature ciliated cells of guinea-pig tracheal epithelium. A few biciliated cells were observed, and in a rare case one cell had developed four such processes. Primary cilia are characterized by a 9 + 0 microtubular arrangement near the base, while a transition to an 8 + 1 pattern occurs at a slightly more distal position. Spokes are lacking, and dynein arms are absent or incompletely developed. The function, if any, of primary cilia remains unknown.In the population of the motile 9 + 2 cilia atypical forms are very rare, i.e. <0.1%. Of the various abnormalities cilia with supernumary microtubules are most common. Only one atypical basal body was observed. Although some of the aberrant forms undoubtedly are non-motile, their very low number suggests that they have no practical influence on the muco-ciliary clearance.Local extrusions of the ciliary membrane, here named ciliary knobs, are believed to be fixation artefacts. It is suggested that they represent circumscribed regions of the ciliary membrane which are sensitive to changes in the environmental osmotic pressure.     

The fine structure of atypical ciliated cells in the human gastric epithelium

Gastric mucosa obtained from the body and pyloric portions of the human stomach were observed by light and transmission electron microscopy. Ciliated cells were found in two of 18 subjects examined, one patient with gastric ulcer and the other one with gastric adenocarcinoma. The ciliated cells were found in epithelia at sites away from the main lesions. The tissues containing ciliated cells showed intestinal metaplasia combined with mild chronic gastritis in both cases. The epithelial layer facing the gastric lumen was composed of columnar cells with numerous uniform microvilli and goblet cells. This epithelium extended to the superficial parts of the tubules surrounded by the lamina propria. The deeper portions of the tubules were composed of mucous secretory, endocrine, and rarely ciliated cells. These ciliated cells were provided with numerous cilia the numbers of which varied considerably from cell to cell. This was in contrast to the primary cilium which is usually single. The central part of the apical cell membrane was sometimes concave in the area from where cilia tended to arise. It was also observed that numerous basal bodies as well as mucus-like granules were contained in the same cell. The axonemal pattern was different from that of ordinary cilia and showed 9 + 0 and 8 + 1 patterns. In longitudinal sections it was found that one peripheral doublet was displaced to the center of the axoneme as it left the basal body.     

Determination of ciliary polarity precedes differentiation in the epithelial cells of quail oviduct.

In quail oviduct epithelium, as in all metazoan and protozoan ciliated cells, cilia beat in a coordinated cycle. They are arranged in a polarized pattern oriented according to the anteroposterior axis of the oviduct and are most likely responsible for transport of the ovum and egg white proteins from the infundibulum toward the uterus. Orientation of ciliary beating is related to that of the basal bodies, indicated by the location of the lateral basal foot, which points in the direction of the active stroke of ciliary beating. This arrangement of the ciliary cortex occurs as the ultimate step in ciliogenesis and following the oviduct development. Cilia first develop in a random orientation and reorient later, simultaneously with the development of the cortical cytoskeleton. In order to know when the final orientation of basal bodies and cilia is determined in the course of oviduct development, microsurgical reversal of a segment of the immature oviduct was performed. Then, after hormone-induced development and ciliogenesis, ciliary orientation was examined in the inverted segment and in normal parts of the ciliated epithelium. In the inverted segment, orientation was reversed, as shown by a video recording of the direction of effective flow produced by beating cilia, by the three-dimensional bending forms of cilia immobilized during the beating cycle and screened by scanning electron microscopy, and by the position of basal body appendages as seen in thin sections by transmission electron microscopy. These results demonstrate that basal body and ciliary orientation are irreversibly determined prior to development by an endogenous signal present early in the cells of the immature oviduct, transmitted to daughter cells during the proliferative phase and expressed at the end of ciliogenesis.     

Scanning electron microscopy of the equine oviduct and observations on ciliary currents in vitro at day 2 after ovulation

There are considerable differences between mammalian species in the distribution and activity of ciliated cells within the oviduct, and limited information is available concerning either the distribution or activity of cilia within the equine oviduct. Patterns of ciliary activity were characterized in the ampulla and isthmus of oviducts recovered at 2 d after ovulation from 10 mares, and scanning electron microscopy was used to examine regional differences in the distribution of cilia in oviducts from 3 of these mares. Based upon the motility of 15 microm latex microspheres, ciliary activity was significantly (P < 0.001) greater in the ampullar oviduct compared with that of the isthmic oviduct. The direction of ciliary beat was consistently toward the uterus in all regions of the oviduct. Scanning electron microscopy revealed ciliated and secretory cells in both regions of the oviduct at 2 d after ovulation, with no apparent differences in the proportion of ciliated versus secretory cells.     

Electron microscopic study of intestinal epithelium of <Emphasis Type="Italic">Saccoglossus mereschkowskii</Emphasis> (Enteropneusta,Hemichordata)

The epithelium of the hepatic region of the intestine in Saccoglossus mereschkowskii, a representative of enteropneusts (Enteropneusta, Hemichordata), a group located at the base of Chordata, has been studied by using electron microscopy. The ultrastructure of ciliated and granular epithelial cells, elements of the intraepithelial nerve layer, and intercellular junctions are characterized. The data on the details of the structure of the ciliary apparatus and the system of ciliary rootlets are presented. Justification is provided for the presence of a complicated construction in the ciliated cells, a supportive carcass of cilia that performs a mechanical stabilizing function, and possibly the synchronization of the ciliary movement. The existence of cilia with two centrioles is considered as adaptation to the high functional load on the ciliary apparatus. Well-developed bundles of myofilaments have been revealed in the cytoplasm of the basal parts of ciliated cells, which characterizes these cells as epitheliomuscular. Peculiarities indicating the role of ciliated cells in absorption are described, as well as the capability of these cells for balloon-like secretion. Data are presented on the accumulation of reserved nutritional substances in the cell cytoplasm in the form of lipids and glycogen. With respect to their function, ciliated cells are determined as the ciliated secretory-absorptive epitheliomuscular cells. The location of secretory granules in both apical and basal parts of granular cells indicates the exocrine-endocrine function of these cells. There are no typical endocrine cells in the intestinal epithelium of S. mereschkowskii. Several types of granules are described in the cytoplasm of nerve fibers. Junctions between nerve fibers and basal parts of ciliated and granular epithelial cells have been revealed; the neural regulation of the contractile and secretory functions of epithelial cells is assumed. The intestinal epithelium of enteropneusts is presumed to contain a regulatory neuroendocrine system composed of receptor cells of the open type, secretory endocrine-like cells, and of nerve elements of the nervous layer.     

解脲支原体对兔输卵管黏膜上皮细胞致病性研究

目的：探讨解脲支原体(UU)对兔输卵管黏膜上皮细胞的粘附及其致病性。方法：采用雌性大白兔作为实验对象,用UU感染兔输卵管黏膜上皮细胞,经光学和电子显微镜观察UU对其粘附性和致病性。结果：UU感染后,兔输卵管管腔有较多渗出物,呈淡红染匀质状,黏膜层的上皮细胞轻度肿胀,黏膜下层有炎症细胞(以淋巴细胞为主)呈灶性浸润;在电镜下,UU粘附于输卵管黏膜上皮细胞上,同时上皮细胞游离面的纤毛互相粘连、倒状,纤毛细胞核膜欠完整．胞核染色质呈凝块状．胞浆内可见大量大小不等的空泡,无纤毛细胞顶面参差不齐,可见伪足样突出．微绒毛减少．胞浆内近细胞顶部有少量分泌颗粒和空泡变性,细胞间可见相嵌连接。结论：UU到达兔输卵管后可粘附于输卵管黏膜上皮细胞上并导致损伤。     

Electron microscopic study of the intestinal epithelium of Saccoglossus mereschkowskii (Enteropneusta, Hemichordata)

Epithelium of the hepatic region of the intestine in Saccoglossus mereschkowskii, a representative of enteropneusts (Enteropneusta, Hemichordata) standing at the base of Chordata, has been investigated using electron microscope. The ultrastructure of ciliated and granular epithelial cells, elements of the intraepithelial nerve layer, and intercellular junctions have been characterized. The data concerning details of the organization of the ciliary apparatus and rootlets system are presented. It is justified the presence of complicated supporting construction of cilia which performs a mechanical stabilizing function and possibly also provide synchronization of ciliary movements. The presence of cilia with two centrioles is considered as an adaptation to high functional load on ciliary apparatus. Well developed bundles of myofilaments are found in the cytoplasm of the basal portions of ciliary cells that characterizes these cells as myoepithelial. The features indicating the role of ciliary cells in absorption are described. The capability of these cells to balloon-like secretion is considered. Data on the accumulation of food reserves in the form of lipid droplets and glycogen in the cell cytoplasm are presented. Ciliated cells are characterized by their function as ciliated secretory-absorptive myoepithelial cells. Based on the location of secretory granules both in the apical and basal portions of granular cells, an exocrine-endocrine function of these cells has been suggested. Typical endocrine cells in the intestinal epithelium of S. mereschkowskii are absent. Several types of granules in the nerve fibers cytoplasm are described. Junctions between the nerve fibers and basal portions of ciliary and granular epithelial cells are found. Nerve regulation of contractile and secretory functions of epithelial cells is supposed. The presence of the regulatory nerve-endocrine system that includes receptor cells of open type, secretory endocrine-like cells and nerve elements of nerve layer is supposed in the intestinal epithelium of enteropneusts.     

The novel centriolar satellite protein SSX2IP targets Cep290 to the ciliary transition zone

In differentiated human cells, primary cilia fulfill essential functions in converting mechanical or chemical stimuli into intracellular signals. Formation and maintenance of cilia require multiple functions associated with the centriole-derived basal body, from which axonemal microtubules grow and which assembles a gate to maintain the specific ciliary proteome. Here we characterize the function of a novel centriolar satellite protein, synovial sarcoma X breakpoint–interacting protein 2 (SSX2IP), in the assembly of primary cilia. We show that SSX2IP localizes to the basal body of primary cilia in human and murine ciliated cells. Using small interfering RNA knockdown in human cells, we demonstrate the importance of SSX2IP for efficient recruitment of the ciliopathy-associated satellite protein Cep290 to both satellites and the basal body. Cep290 takes a central role in gating proteins to the ciliary compartment. Consistent with that, loss of SSX2IP drastically reduces entry of the BBSome, which functions to target membrane proteins to primary cilia, and interferes with efficient accumulation of the key regulator of ciliary membrane protein targeting, Rab8. Finally, we show that SSX2IP knockdown limits targeting of the ciliary membrane protein and BBSome cargo, somatostatin receptor 3, and significantly reduces axoneme length. Our data establish SSX2IP as a novel targeting factor for ciliary membrane proteins cooperating with Cep290, the BBSome, and Rab8.     

Isolation and characterization of cholangiocyte primary cilia

Primary cilia are distinct organelles expressed by many vertebrate cells, including cholangiocytes; however, their functions remain obscure. To begin to explore the physiological role of these organelles in the liver, we described the morphology and structure of cholangiocyte cilia and developed new approaches for their isolation. Primary cilia were present only in bile ducts and were not observed in hepatocytes or in hepatic arterial or portal venous endothelial cells. Each cholangiocyte possesses a single cilium that extends from the apical membrane into the bile duct lumen. In addition, the length of the cilia was proportional to the bile duct diameter. We reproducibly isolated enriched fractions of cilia from normal rat and mouse cholangiocytes by two different approaches as assessed by scanning electron, transmission electron, and confocal microscopy. The purity of isolated ciliary fractions was further analyzed by Western blot analysis using acetylated tubulin as a ciliary marker and P2Y(2) as a nonciliary cell membrane marker. These novel techniques produced enriched ciliary fractions of sufficient purity and quantity for light and electron microscopy and for biochemical analyses. They will permit further assessment of the role of primary cilia in normal and pathological conditions.     

Electron microscopic observations on tracheal epithelia of mice infected with Bordetella bronchiseptica

To clarify the pathogenesis of Bordetella in vivo infection, the tracheal epithelia of mice were examined in detail by electron microscopy at various intervals after intranasal inoculation with graded doses of phase I Bordetella bronchiseptica. In mice infected with a lethal dose (6 to 7 x 10(7) CFU), a remarkable rupture of the cell membranes of cilia and microvilli of the middle trachea was found on day I postinfection. The rupture of the membrane was observed over the entire tracheal epithelia, on day 2 after infection. The affected cilia were constricted at the transitional region and were broken off. In the ciliated cells the adherence of organisms to ciliary apexes and colonization in the interciliary spaces were also remarkable. In both the ciliated and nonciliated epithelial cells, the cytoplasmic vacuolation and pyknosis or karyorrehexis were also notable. In mice infected with one-tenth of the lethal dose, similar findings were seen, but appeared more slowly and the bacteria were not seen attaching to ciliary apexes. In mice receiving one-hundredth of the lethal dose, only mild cilial abnormality such as aggregation of cilia, and slight cytoplasmic vacuolation were found 6 days postinfection. Based on these findings, a possible mechanism of the ciliary damages produced by B. bronchiseptica was postulated.     

Some aspects of the fine structure of the statocysts of the molluscsPecten andPterotrachea

Summary The statocyst ofPecten is composed of hair cells and supporting cells. The hair cells bear kinocilia and microvilli at their distal ends and the supporting cells bear microvilli. The cilia have a 9+2 internal filament content, and arise from basal bodies that have roots, basal feet and microtubular connections. Two different ciliary arrangements are described, one with a small number of cilia arranged in a ring, and another with many more cilia arranged in rows. Below the hair cells are probable synapses. A ciliated duct connects to the lumen of the static sac and passes through the centre of the static nerve. The hair cells in the statocyst ofPterotrachea bear kinocilia and microvilli. The possible importance of cilia and microvilli in the transduction process is discussed.We would like to thank ProfessorJ. Z. Young for bringing specimens ofPterotrachea from Naples and also the staff of the Stazione Zoologica for the provision of specimens, Dr.M. Land for providing specimens ofPecten, the Science Research Council (U.K.) for providing the electron microscope used in much of the study and also for a grant to one of us (V.C.B.), and Mrs.J. Parkers and Mr.R. Moss and Mrs.J. Hamilton for much photographic and technical assistance.     

Functional activity of ciliated outgrowths from cultured human nasal and tracheal epithelia

Primary cultures of respiratory epithelium were produced as outgrowths from human fetal and adult tracheal and nasal polyp explants. Video recordings of the epithelial cell outgrowths were carried out after 5 days of culture and the ciliary beating frequency was analyzed by using a video technique. Uniform fields of differentiated ciliated cells were observed near the edge of the explant. In the transition region of the outgrowth from the explant to the outgrowth periphery, isolated ciliated cells were present, as well as cells with fused cilia. The ciliary beating frequency of the outgrowth of well-differentiated ciliated cells (13.5 +/- 1.4 Hz) was significantly higher (p less than 0.001) than the beating frequency of both the explant (11.9 +/- 0.7 Hz) and the ciliated cells with fused cilia (9.8 +/- 1.7 Hz). The same differentiation stages and functional activities were observed in the outgrowth cultures, whatever their origin. These in vitro models are comparable with each other and therefore could be useful for studying the ciliogenesis and functional activity of the human respiratory epithelium.     

Ciliated band structure in planktotrophic and lecithotrophic larvae of Heliocidaris species (Echinodermata: Echinoidea): a demonstration of conservation and change

The evolution of lecithotrophic (non-feeding) development in sea urchins is associated with reduction or loss of structures found in the planktotrophic (feeding) echinopluteus larvae. Reductions or losses of larval feeding structures include pluteal arms, their supporting skeleton and the ciliated band that borders them. The barrel-shaped lecithotrophic larva of Heliocidaris erythrogramma has, at its posterior end, two or three ciliated band segments comprised of densely packed, elongate cilia. These cilia may be expressions of the epaulettes that would have been present in an ancestral larval form, represented today by the feeding echinopluteus of H. tuberculata . We compared the development and cellular organization of the larval ciliary structures of both Heliocidaris species to assess whether the ciliary bands of H. erythrogramma are expressions of the feeding ciliated band or epaulettes of an echinopluteus. Epaulette development in feeding larvae of H. tuberculata involves separation of specific parts of the ciliated band from the rest of the feeding ciliated band, hyperplastic addition of ciliated cells and hypertrophic growth of the cilia. Like epaulettes, the ciliated bands of H. erythrogramma are composed of long spindle-shaped cells arranged in a cup-shaped collection that bulges into the blastocoel; and these cells have elongated cilia. In their developmental origin and topological arrangement however, the ciliated bands of H. erythrogramma correspond more closely with parts of the pluteal feeding ciliated band than with epaulettes. The larvae of this echinoid appear to develop epaulette-like bands from parts of the original (but reduced) feeding ciliated band. The evolution of development in H. erythrogramma has thus involved both conservation and change in echinopluteal ciliary structures.     

Scoring a backstage pass: mechanisms of ciliogenesis and ciliary access

Cilia are conserved, microtubule-based cell surface projections that emanate from basal bodies, membrane-docked centrioles. The beating of motile cilia and flagella enables cells to swim and epithelia to displace fluids. In contrast, most primary cilia do not beat but instead detect environmental or intercellular stimuli. Inborn defects in both kinds of cilia cause human ciliopathies, diseases with diverse manifestations such as heterotaxia and kidney cysts. These diseases are caused by defects in ciliogenesis or ciliary function. The signaling functions of cilia require regulation of ciliary composition, which depends on the control of protein traffic into and out of cilia.     

Patterns of basal body addition in ciliary rows in Tetrahymena

Most naked basal bodies visualized in protargol stains on the surface of Tetrahymena are new basal bodies which have not yet developed cilia. The rarity of short cilia is explained by the rapid development of the ciliary shaft once it begins to grow. The high frequency of naked basal bodies (about 50 percent) in log cultures indicates that the interval between assembly of the basal body and the initiation of the cilium is long, approximately a full cell cycle. Naked basal bodies are more frequent in the mid and posterior parts of the cell and two or more naked basal bodies may be associated with one ciliated basal body in these regions. Daughter cells produced at division are apparently asymmetric with respect to their endowment of new and old organelles.     

Single modified cilia displayed by cells of human internal stratified epithelia (oral cavity,vagina)

Summary Serial sections of human vaginal and keratinized oral-gingival epithelia were investigated for ciliary structures. Most melanocytes of the gingival epithelium lacked cilia, whereas almost all basal keratinocytes of the deeper portion of the epithelial ridges possessed one cilium each. In the suprabasal layers of the ridges only a few keratinocytes exhibited a single cilium. In the basal layer, at the top of the connective tissue papillae, approximately every second keratinocyte displayed a single cilium. In the suprabasal layers above the ridges no ciliated keratinocytes were observed. The basal cells of the vaginal epithelium were endowed with cilia, while cilia were absent from the suprabasal cells. In the human forearm epidermis most melanocytes and keratinocytes are supplied with a single cilium; it has been suggested that they may play a role in light reception. However, the widespread occurrence of 9 + 0 cilia in epithelial cells of internal epithelia and their coincidence with the sites of renewal of keratinocytes suggests that a relationship may exist between solitary cilia and mitotic activity.