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On the differentiation of prospective ectoderm to a ciliated cell pattern in embryos of Ambystoma mexicanum.

The differentiation of the ectoderm in Ambystoma mexicanum (Harrison stage 26--27) was examined under in vivo and in vitro conditions by scanning electron microscopy under different experimental conditions. About one out of three flank epidermal cells was found to be ciliated in the undistrubed or control embryos. The shape of ciliated cells in the explants from the animal region was only slightly affected. In no case was it possible to find two adjacent ciliated cells, implying that these cells prevent the appearance of cilia in the cells in direct contact. Transformation to ciliated cells is suppressed by hypertonicity but favoured in a hypotonic medium. The differentiation of epidermis is also dependent upon the synthesis of RNA and some kind of sulphated glucosaminoglycan, corroborated by the inhibitory effect of actinomycin and selenate. The differences between the test series and the controls are discussed with regard to factors controlling embryonic epidermal differentiation.

Ambystoma

The cyclical variation in the percentage of ciliated cells in the normal human endometrium.

The percentage of ciliated cells in the luminal and glandular epithelia of endometrial samples from sixty-eight normal women has been studied. Although the concentration of ciliated cells found in the luminal epithelium tended to lag behind and below those found in the glandular epithelium, no significant difference was found between the absolute percentages of ciliated cells in each site. The number of ciliated cells increased during the proliferative phase to reach a maximum of around 20%. This was maintained during the ovulatory phase, and then declined. The hormonal basis of this variation is discussed.

Adolescent

[Ciliated cells of corneal endothelium. Functional and morphological aspects compared to cilia of other organs].

The presence of ciliated cells has been demonstrated by scanning and transmission electron microscopy in corneal endothelium. This study has been carried out on human cornea in normal and pathological cases and on rabbit's normal cornea. Another study has been lead on regeneration of endothelium after freeze destruction. The cilia cells described have a normal structure. The cilia has nine peripheral groups of tubules without any tubule in the center. The cilia seem to be more numerous in case of corneal disease and during the regeneration of corneal endothelium. The authors compare the presence of ciliated cells in different tissues. This suggest that the potency to form single cilia is present in almost every kind of tissue but there is no proof of their pathological importance. In the corneal endothelium, as in the kidney, cilia seem to have no function of fluid transportation but the possibility for cilia to be a receptor is discussed.

Animals

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

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

Adenosine Triphosphatases

[Triggering and regulatory mechanisms of ciliary motion in the ctenophore Bolinopsis. III. Electric excitability and inherent contractility of ciliated cells].

Rhythmical electrostimulation is able to produce in a tissue strip of Bolinopsis series of beats of combs-plates. However, the beat frequency, within one series, is only poorly controlled by electrical stimuli. Isolated comb-plate cells with a cilium cut off maintain rhythmical contractions stimulated both mechanically and electrically. A lot of microtubules has been found in the apical region of these cells. It is supposed that it is these microtubules, probably together with the associated root filaments, that are responsible for cell motility. Mutual mechanical stimulation of ciliary cells in these motions is, presumably, a signal providing intratissue propagation of metachronal wave.

Animals

The respiratory epithelium. I. Human bronchus.

Six morphologic cell types comprise the human bronchial epithelium: basal cells that do not reach the bronchial lumen, neurosecretory cells (Kulchitsky's cells, K-cells, or small granule cells) that rarely reach the lumen, and indifferent cells, mucous cells [small mucous granule cells (SMGC) and mucous goblet cells], ciliated cells, and ciliated-mucous cells that do reach the lumen. Ciliated-mucous cells bearing fully developed cilia and containing mucous granules are seen only occasionally. Three of the cell types that reach the lumen are microvillus covered and do not bear cilia. The microvillus-covered nonciliated cells are: 1) neurosecretory cells, 2) indifferent cells, and 3) mucous cells. Neurosecretory cells contain characteristic dense core granules. Such cells rarely reach the lumen. Indifferent cells are rarely seen. They have a pale cytoplasm and show no evidence of either ciliary or mucous differentiation. Similar cells are observed showing early signs of either ciliary or mucous differentiation or even both types of differentiation in the same cell. Mucous cells comprise the vast majority of microvillus-covered cells. They present either as SMGC with a few small mucous granules or as goblet cells, filled with mucus. These columnar cells are characterized ultrastructurally by dense cytoplasm and a well-developed endoplasmic reticulum and Golgi apparatus. The microvilli are coated with a glycocalyx that binds colloidal iron more avidly than that of either cilia or microvilli of ciliated cells. Possible interrelationships between the different cell types in normal epithelium are discussed.

Bronchi

New ultrastructural features on the cream hamster thyroid with special reference to the second kind of follicle.

The thyroid cells of the cream hamster, characterized by abundance of microtubules and stratification of the organelles, undergo a particular evolution when the animals grow older. These changes are characterized by an increase of the number of lysosomes which in extreme cases become so prominent that they occupy the whole cytoplasm of the cell which thus loses its organelle stratification. As in other species, cream hamster thyroid contains so-called ultimobranchial follicles made up of at least six cell types: fibrillar dark and light cells, parafollicular cells, ciliated cells, vesicular cells, and cells with myelinic inclusions. The ultrastructure of these follicles in the cream hamster represents a mixture of the ultrastructural characteristics of the same follicles encountered in the rat and the mouse thyroid. Here also mixed follicles are seen. Nevertheless vesicular cells present such abundant "secretion granules" that the question arises as to whether these follicles produce a special secretion and perhaps a new hormone. Incubation of cream hamster thyroids in the prescence of vincristine induces vanishing of microtubules, formation of paracrystalline structures, and loss of stratification of the organelles. Although these last effects might be due to some specific toxic effect of the drug, it is suggested that the disappearing of the organelle stratification might result from a specific vincristine-induced disaggregation of the microtubules acting as a cytoskeleton.

Age Factors

[The fine structure of the epithelium of the main outlets from the liver and pancreas of the domestic chicken (author's transl)].

Light- and electronmicroscopic and some histochemical investigations on the major excretory ducts of pancreas and liver was carried out on 24 adult chicken. The epithelium of both ducts was shown to include: cells with apical secretory granules, non-differenciated duct cells, ciliated cells and migrating cells. In addition the epithelium of the pancreatic ducts included goblet cells. The distal plasmalemm and the microvilli are covered with surfacecoat, which like the secretory granules produces precipitate with PA-silver. This area of the cell also shows a strong reaction to M++ and Ca++ activated ATPase.

Adenosine Triphosphatases

Scanning electron microscopy of human female reproductive tract and amniotic fluid cells.

Scanning electron microscopy was used to examine surface ultrastructural characteristics of cells of the epithelium of female reproductive tract, cervical mucus, and amniotic fluid cells. The female epithelium undergoes hormone-dependent cyclical morphological alterations in cell shape, apical microvilli, ciliation, and secretory patterns. The frequency distribution of ciliated cells varies in different parts of the female reproductive tract and different segments of the same organ. In the endocervix, a few ciliated cells are randomly distributed. The density of ciliated cells is variable in several areas of the endometrium. The sparse distribution of ciliated cells in the central portion of the uterine wall may indicate a possible implantation site. According to surface ultrastructure of cells, three segments are noted in the uterotubal junction: cornual endometrium, interstitial tubal epithelium, and transitional area. The vaginal epithelium is made of flat polygonal cells with interdigitating borders and a fine interlacing network of microridges. After the menopause, the microridge pattern and distinct cell borders are lost or severely modified. Two types of fibrous structures can be found in cervical mucus: a) microfibrils that vary in diameter from 500 to 1,500 A and from bundles or networks, and b) long, thick fibers that vary in diameter from 0.5 to 5 micrometer run parallel to each other, and appear to be made up of microfibrils. Amniotic fluid cells, derived from amniotic epithelium and fetal ectoderm, consist of non-nucleated and nucleated cells with extensive microplicae.

Amniotic Fluid

The respiratory epithelium. II. Hamster trachea, bronchus, and bronchioles.

The normal female hamster respiratory epithelium at five airway levels was characterized with the use of coordinated morphologic and histochemical techniques. Five morphologic cell types were recognized in the trachea, stem bronchi, and primary bronchl: basal cells and neurosecretory cells that were basally located and did not reach the lumen and mucous cells [mucous goblet cells and small mucous granule cells (SMGC)], indifferent cells showing mucous-ciliary differentiation, and ciliated cells that reached the lumen. Two epithelial cell types were observed in the bronchioles, ciliated cells and nonciliated Clara cells, both of which reached the lumen. Mucous cells presented as either SMGC with a few small periodic acid-Schiff-positive granules (diastase-resistant neutral mucosubstances) or as goblet cells, filled with the same material. Mucous cells were columnar, and the cytoplasm was electron-dense and contained a well-developed endoplasmic reticulum and Golgi complex. The microvilli of the mucous cells were coated more thickly with colloidal iron than either the cilia or microvilli of ciliated cells. Approximately one-half the cells in the trachea, bronchi, and bronchioles were ciliated. Ciliated cells containing intracellular ciliated cysts with normal cilia projecting into a closed space or ciliated cells bearing compound cilia were observed infrequently. Neurosecretory cells were rarely observed. These cells contained characteristic dense-core granules.

Animals

Respiratory tract epithelium in primary culture: concurrent growth and differentiation during establishment.

Studies of cellular function in the respiratory tract lining have traditionally been limited by the small tissue mass and functional diversity of the epithelium. Recent improvements in culture conditions have permitted long-term maintenance of epithelial cells derived from the upper respiratory tract of rats. The present study determined the extent to which proliferation and differentiation took place in such epithelial cell cultures. The labelling index obtained after 3H-dThd administration was approximately 100-fold higher than that of the quiescent epithelium in vivo; therefore, a large proportion of the cells were probably in the cycling population. Ultrastructural studies showed that features unique to the specialized mucous secretory cells and ciliated cells were lost rapidly with entry of these cells into the in vitro environment. With long-term maintenance, the cultures were reorganized into a stratified epithelium containing large, squamous, apical cells and small basal cells. The ultrastructural appearance of basal cells in vitro was nearly identical to that of basal cells in vivo. Squamous cells were frequently joined by tight junctions. Because hemicysts originated by detachment of squamous cells from the basal layers but not from adjacent squamous cells, they were considered to indicate stratification in the cultures. The proliferative and differentiative status of the mucociliary epithelium was altered by in vitro conditions, and came to resemble that of early squamous metaplasia in the respiratory tract epithelium.

Animals

Cell ultrastructure during the development and the transformation of a pig--uterine tube cell line.

Observations by electron microscopy disclosed ultrastructural features which were not apparent by light microscopy during the three stages of the development of a pig uterine tube cell line. Ciliated cells without dynein arms were found naturally occurring in the first 100 subcultures (pseudo-diploid stage). They were not observed in the routinely growing cultures after the concomitant detection of the first chromosome marker and expression of a type C virus, unless the cells were treated with estradiol (transformed stage). Gap junctions and increased numbers of lamellar and lysosomic bodies and vacuole-like structures were observed in clusters of tumoral cells in the high subcultures if a new chromosome marker occurred (malignant stage). Type C viruses were found as early as the 106th subculture (transformed stage) in populations of actively dividing cells 3 days after seeding, but not in populations of nondividing cells a week after seeding. Both viruses and viral reverse transcriptase were continuously released in the 300 subsequent subcultures.

Animals

[Histologic data on the lingual glands in various anuran amphibians].

The anuran Amphibians have a well developed tongue which is either protractile or attached to the floor of the buccal cavity according to the species. The lingual glands of the papillary dorsal side consist of simple tubules. The stratified epithelium of the ventral and lateral sides of the tongue is composed of mucous and muco-serous cells inserted between the covering ciliated cells. Filiform pappillae and glands have a simple epithelium with numerous outer ciliated cells and glandular cells. The latter eleborate a product rich in glycosaminoglycans, more or less acid according to the species, associated with proteins. The lingual gland secretion is composed of proteins; when present, glycosaminoglycans are less abundant and less acid than those produced by the outer glandular cells. The histochemical and ultrastructural characters of the lingual glands of Anurans are compared to those of Apoda and Caudata previously studied by the same techniques.

Amphibians

Calcium control of ciliary arrest in mussel gill cells.

After several hours in 20 mM sodium phosphate and 40 mM KCI (pH 7.4) or similar simple solutions, ciliated cells exfoliate en masse from stripped gill epithelium of freshwater mussels, e.g., Elliptio complanatus. Three types of ciliated cells--lateral (L), laterofrontal (LF), and frontal (F)--can be distiniguished and counted separately in the suspensions. About one-half of the cells of each type remain motile. Motility is unaffected by addition of 10(-5) M A23187 or 10(-2) M Ca+2 added separately, but when ionophore and Ca+2 are added together, ciliary beat is largely arrested. Treatment of the cells with Triton X-100 (Rohm & Haas Co., Philadelphia, Pa.) results in a total loss of motility as the ciliary membrane becomes disrupted. Such models can be reactivated by addition of ATP and Mg+2. All ciliated cell types are reactivated to about the same extent. At least 80% of the activity of the untreated preparation returns. Ca+2-EGTA buffers added to the reactivating solutions permit titration of free Ca+2 concentration vs. percent motility. Activity is unchanged for all cell types at Ca+2 less than 10(-7) M; at 10(-6) Ca+2, L cilia of all cell types are arrested differentially, whereas at Ca+2 greater than 10(-4) M most cilia of all cell types are arrested. We conclude: (a) that increasing cytoplasmic Ca+2 is directly responsible for ciliary arrest, (b) that the readily reversible physiological arrest response of the L cilia in the intact gill is caused by a rise in free Ca+2 in narrow limits from ca. 5 x 10(-7) M to ca. 8 x 10(-7) M, and (c) that the site which is sensitive to Ca+2 is part of the ciliary axoneme or the basal apparatus.

Adenosine Triphosphate

[Ultrastructure of "villose cells" in the early stages of their formation in the process of chemical carcinogenesis in the CNS].

Early stages of redistribution of cellular elements around the pill of carcinogenic agent DMBA introduced into the right hemisphere of the brain of female SHK albino rats were studied. Precursors of the ciliated cells were established to appear in the perivascular tissue within 12 h. Within 24 h they accumulate around the pill bed, and within 48 h a cellular edging is formed around DMBA. The cytoplasm of ciliated cells become richer in organelles. The main marker in identification of cells in all periods of experiment were lipoid inclusions in the cytoplasm which are different from lipids of usual macrophages occurring both in experiments and in control, in a polygonal shape and sinuous contour. Within 48 h the cytoplasm of ciliated cells form long lancet-shaped spiculae with upright walls. The cytoplasm of macrophages gives only short, somewhat sinuous processes.

9,10-Dimethyl-1,2-benzanthracene

Fetal mouse lung in circumfusion system cultures.

Fetal mouse lungs were cultivated, using the dual-rotary circumfusion system for tissue culture, and their histotypic development was surveyed for 75 days by phase-contrast and electron microscopy. Alveoli, terminal bronchioles and alveolar macrophages were photographed periodically with still and time-lapse phase-contrast microscopy. Their histotypic appearance was confirmed by electron micrographs of the 1- and 2 1/2-month-old specimens. These revealed typical alveoli surrounded by a basal lamina and composed of types I and II pneumocytes containing various lamellar-body forms within the type II cells, the alveolar lumen, and the alveolar macrophages. There was a shift from almost all type II cells in the 1-month-old alveoli to the presence of frequent type I cells as constituents of the alveoli in the 2 1/2-month-old cultures. The terminal bronchioles were tubules consisting of ciliated cells with Clara cells interspersed between them. The ciliated cells contained as many as 30 cilia or basal bodies per section and numerous microvilli. They were attached to each other and to the Clara cells by junctional complexes and accessory desmosomes which were generally in the apical ends of the cells. The Clara cells typically had glycogen granules interspersed between lamellae of the endoplasmic reticulum, contained numerous well dispersed mitochondria, occasional lysosome-like granules and crystalloid bodies which appeared to be tubular. Some Clara cells presented a moderatley dense secretory granule in the center of the whorl of the endoplasmic reticulum.

Animals

Origin of ciliated alveolar epithelial cells in bleomycin-induced lung injury.

Bleomycin is known to induce diffuse pulmonary fibrosis and epithelial metaplasia. The reaction of the alveolar epithelium following a single intravenous or multiple intraperitoneal injections of bleomycin to mice is now examined in a combined morphologic and cytodynamic study. Necrosis of Type 1 cells was observed, followed by proliferation of Type 2 cells, a common reparative process. The proliferated cells transformed to a variety of epithelial forms, including ciliated cells and cells with morphologic features intermediate between alveolar and bronchiolar epithelium. No evidence of cell injury or increased cell division was found in the bronchial epithelium. It is concluded that the metaplastic ciliated epithelial cells are produced by an abnormal reparative process in the alveolar epithelium. The results suggest that, whereas the "resting" Type 2 cell is not vulnerable to bleomycin, in the postmitotic phase the drug may modify the synthetic mechanisms of cellular differentiation and thereby induce metaplasia.

Animals

Cellular imbalance in proximal and distal lung of CFTR-/- sheep in utero and at birth.

BACKGROUND: The Lung is the major focus of therapeutic approaches for the inherited disorder cystic fibrosis (CF) as without treatment lung disease is life-limiting. However, the initiating events that predispose the CF lung to cycles of infection, inflammation and resultant tissue damage are still unclear. Inflammation may occur in the CF lung prior to birth in human and several large animal models suggesting an in utero origin for the disease and encouraging further studies prior to birth. METHODS: Here we used the sheep model of CF (CFTR-/-) and age-matched wild-type (WT) sheep of the same breed to investigate the single cell transcriptomes of proximal and distal lung tissue at 80 days and 120 days of gestation and at term (147 days). Single cell RNA-seq was performed on tissues from 4 to 7 animals of each genotype (WT and CFTR-/-) at each time point. RESULTS: At term, FOXJ1-expressing ciliated cells are overrepresented in both lung regions from CFTR-/- lambs, while secretory epithelial and basal cells are underrepresented in proximal lung, as are T cells and monocytes in distal lung. The imbalance in ciliated and basal cells was confirmed by immunohistochemistry. At 120 days of gestation, lymphoid cells are slightly more abundant in proximal and distal lung from CFTR-/- animals compared to WT, consistent with the transient CF-associated inflammatory response in utero. At 80 days of gestation, T and B cells are underrepresented in both lung regions. CONCLUSIONS: The differences in epithelial cell abundance observed in the CFTR-/- lambs at term may reflect sequelae from the loss of CFTR on lung development and differentiation in utero. These findings provide novel insights into the cellular mechanisms of pathology and may be relevant to the design of new therapeutic approaches for CF lung disease.

Animals