Gap junctions and impulse propagation in embryonic epithelium of Amphibia

Summary Epithelium of amphibian embryos (Cynops orientalis, Xenopus laevis) was found in preceding experiments to generate and conduct impulses during a limited stage (26–37) of development. In order to elucidate the structural basis of impulse propagation, epithelial cells of four stages were examined by the freeze-etching method: (I) before and (II) during acquisition of conductivity; (III) when propagation was fully established, and (IV) when it was no longer present. Only few gap junctions (GJ) of small size were found in groups I and IV. GJ in epithelia of group III were increased in number and size, and appeared morphologically coupled, i.e., with more loosely arranged connexons. The size of gap-junctional particles did not differ significantly between coupled and uncoupled stages. Zonulae occludentes seemed leaky in stage I, and tight in stages II–IV. Thus, the morphological characteristics of specialized junctions between non excitable cells correlated with the opening and closing of low resistance intercellular current pathways during embryonic development.Gap junctions in particular seem to form an essential link in the non-neural stimulus-response system, which may facilitate the mobility of the embryo during early phases of aquatic life before the reflex pathways have been established. Coupling and uncoupling of gap junctions may also play an important role in the regulation of cell differentiation and morphogenetic movement. The experimental model used in this study provides a useful tool for further investigations of structural correlates of gap junctional permeability under physiological conditions.     

Development of the ovarian surface and associated germ cells in the human fetus

Summary The ovarian surface and associated germ cells have been studied in human fetuses from 12 weeks of age until near term, using light, TEM and SEM techniques. The surface epithelium and related cords proliferate extensively, especially at midterm. The cords in the ovarian cortex appear to be linked with ingrowths from the surface epithelium, and both structures have a common basal lamina. Germ cells are always interspersed among the somatic cells of the surface epithelium and associated cords. These results indicate that both the proliferating cords and surface epithelium may contribute to the formation of early follicles. Furthermore, the occurrence, of elements having some of the features of primitive steroidogenic cells in the regions of cordsurface epithelium continuity, suggests that both structures (surface epithelium and cords) contribute somatic cells, which in addition to becoming granulosa cells, might also contribute to the provision of primitive interstitial cells.Gonocytes tend to migrate through the developing ovarian tissue towards the surface where they become extruded into the peritoneal cavity. This phenomenon might contribute to the reduction in the number of germ cells at birth and parallels the atretic processes within the ovary.     

Morphology and lactose synthesis in tissue culture of mammary alveoli isolated from lactating mice

Summary Mammary epithelial cells from lactating mice synthesize and secrete lactose in culture and retain many features of their in vivo morphology if mammary glands are only partially dissociated to alveoli, rather than completely dissociated to single cells. After 5 d in culture lactose synthesis by alveoli cultured on floating collagen gels is 10 to 20 times higher than in cultures of single cells on floating collagen gels. Moreover, mammary alveoli in culture retain sensitivity to lactogenic hormones; the synthesis of lactose by alveoli depends on the continued presence of insulin and either hydrocortisone or prolactin. In addition, within alveoli the original juxtaposition of constituent epithelial cells is retained, and cells are cuboidal and have many microvilli and fat droplets. In contrast, alveoli on attached gels flatten and lose their secretory morphology. These results indicate that the shape of the cells, presence of lactogenic hormones, and maintenance of epithelial:epithelial cell contacts are required for maintenance of mammary epithelial cell differentiation in culture. This research was supported by Grants CA-16392 and AG-02909 from the National Institutes of Health and Institutional Grant IN 119 from the American Cancer Society.     

Scanning and transmission electron microscopy of the squamose gill-filament epithelium from fresh- and seawater adaptedTilapia

Synopsis The architecture of the gill structure of variousTilapia species was studied in relation to their adaptability to hypersaline media. Using SEM and EM, it was shown that the squamose epithelial cells of the gills have species-typical patterns of ridges on their outer surfaces. These have previously been misinterpreted by other authors as microvilli or stereocillia. The ridges are more dense and better developed in euryhaline species, likeT. zillii, and less so in stenohaline species likeSarotherodon niloticus. Comparing freshwater and seawater-adapted individuals ofT. zillii, S. niloticus, S. galflaeus, andTristramella sacra, it was shown that in fresh water the surface cells are slightly swollen, extending over the openings of the chloride cells. During adaptation to sea water, these ridges become higher and denser and the cell surface shrinks, exposing the underlying orifices of the apical crypts of the chloride cells. The more euryhaline the species, the less change there is in the ridge pattern of the cells during passage from fresh to sea water. This evidence implicates the gill epithelium, together with the chloride cells, in the process of osmoregulation.     

Scanning electron microscopy of the disruption of tobacco hornworm, Manduca sexta, midgut by Bacillus thuringiensis endotoxin

The ingestion of Bacillus thuringiensis crystal endotoxin by Manduca sexta causes the destruction of both goblet and columnar cells of the midgut. One hour after ingestion, the microvilli show pathological effects. Nearly complete destruction of the goblet and columnar cells has taken place after 4 hr exposure to the toxin.     

Human vaginal epithelial multilayer tissue culture

Summary Fragments of normal human adult vagina, when explanted onto glass slides gave rise to outgrowing sheets of pure epithelium, which had microscopic morphological features in common with normal vaginal epithelium. Infrequent fibroblast contamination was observed. Proliferating epithelial cells formedmultilayers of stratified squamous epithelium and demonstrated a progressive decrease in proliferative activity after 14 days. Continuous lines of epithelial cells were not obtained. Even in the absence of estrogens, transmission electron microscopy revealed evidence of keratinization of the superficial cells of the multilayer. Scanning electron microscopy of the surface of mature epithelial cells in culture revealed ultrastructural features that closely resembled those present on the surface of exfoliated cells obtained by scraping the vagina in vivo. This in vitro tissue culture model of human vaginal epithelium may provide a simple method of studying factors that influence vaginal epithelium growth, maturation and function.     

Ultrastructure of the salivary bladder of the nine-banded armadillo

Summary The nine-banded armadillo possesses a salivary bladder which is a dilated portion of the main duct of the submandibular gland at its origin. The wall of the bladder is composed of an epithelium, a submucosa and a thick coat of skeletal muscle. The ultrastructure of the epithelium reveals that it is complex and consists of three cell types: 1) principal cells, 2) light cells, and 3) basal cells. The general organization of the epithelium suggests that it is a transporting type of epithelium such as that found in the amphibian and reptilian urinary bladders and the mammalian gall bladder. The submucosa is composed primarily of densely-packed collagen fibers. The skeletal muscle is very vascular and richly innervated.This study was supported in part by a research grant from U.S.P.H.S. (GRS 5-S01-RR-05705)The authors wish to acknowledge the technical assistance of Elizabeth Underwood     

The occurrence of argyrophilic and argentaffin cells in the gut of Ciona intestinalis L.

Summary Argyrophilic and argentaffin cells occur in the stomach and intestinal epithelium of the sea-squirt, Ciona intestinalis L.. These cells are characterized by their basal swelling which contains the nucleus surrounded by small secretory granules and by a filamentous cell-apex which reaches the gut lumen. The cells are scattered unevenly within the epithelium. Their number decreases rapidly towards the lower part of the intestine. The localization, size of granules and their shape are features which differentiate these cells from other secretory cells in the gut epithelium such as mucous cells. These cells are thought to possess an endocrine function.The excellent technical assistance of Mrs. R. Sprang is gratefully acknowledged     

The fine structure and histochemistry of the caecal epithelium of Calicotyle kröyeri (Monogenea: Monopisthocotylea)

The caecal epithelium of Calicotyle kröyeri consists of a single cell type which functions in the uptake and intracellular digestion of host epidermis and associated mucus. Each cell is columnar with a small basal nucleus and prominent nucleolus. Perinuclear cytoplasm contains narrow profiles of GER and mitochondria with numerous cristae. Golgi complexes are small and indistinct. Most of the cell is filled with vacuoles of heterogeneous content, the largest occupying the cell apex. There is in each cell an apical endocytotic complex comprising cell surface lamellae, apical vesicles and numerous tubular invaginations of the plasmalemma. The limiting membrane of all these components is structurally modified and bears a highly organized array of peg-like structures on its luminal surface. The complex is capable of ingesting particulate food material from the gut lumen for transfer, via vesicles, to the vacuoles for digestion. Most of the vacuoles represent the digestive elements of the cell and, histochemically, are reactive for protein, mucus and carboxylic esterases. Indigestible residues and lipid droplets accumulate in the large apical vacuole and are periodically released to the lumen by exocytosis. Small, undifferentiated caecal cells were occasionally observed in the epithelium, but their development has not been recorded.