A quantitative electron microscopic analysis of the keratinizing epithelium of normal human hard palate

Summary The epithelium of normal human hard palate was subjected to stereologic analysis. Ten biopsies were selected from a total of twenty specimens collected from 9 to 16 year old females, and processed for light- and electron microscopy. At two levels of magnification, electron micrographs were sampled from three strata (basale, spinosum, granulosum) in two locations (epithelial ridges and portions over connective tissue papillae). Stereologic point counting procedures were employed to analyse a total 1560 electron micrographs. In general, the thickness of the palate epithelium was 0.12 mm (over papillae) and 0.31 mm (in ridges), the epithelium is distinctly stratified, and homogeneously ortho-keratinized. From basal to granular layers, the composition of strata revealed decreasing densities of nuclei, mitochondria, membrane-bound organelles and aggregates of free ribosomes. Keratohyalin bodies and membrane coating granules increased, and cytoplasmic filaments with a constant diameter of about 85 Å increased from 14 to 30% of cytoplasmic unit volume. The cytoplasmic ground substance occupied a stable 50% of the epithelial cytoplasm in all strata. The composition of basal layers in ridges differed from that over connective tissue papillae. The data are discussed in relation to the observations that (1) an increasing gradient of filament density is not the most characteristic feature of ortho-keratinizing oral epithelium and (2) differences in the degree of differentiation in cells of the stratum basale coincided with the comparable frequency distribution pattern of dividing cells.The authors are thankful to Miss K. Rossinsky for excellent technical assistance, to Mrs. M. Graf-de Beer for competent data computation and to Mrs. S. Münzel-Pedrazzoli for help in morphometric analysis. This study was in part supported by Grants Nos. 51 and 106 of the Hartmann Müller Foundation and by a Grant from the Foundation of Scientific Research at the University of Zürich.     

Epithelial differentiation at the edentulous alveolar ridge in man

Summary The epithelial lining of the mucosa of the edentulous, maxillary alveolar ridge was subjected to an ultrastructural and stereological analysis. Four biopsies collected from the non-inflamed crest, i.e., the center over former tooth sockets, in non-denture-wearing female patients 30 to 55 years of age were processed for light and electron microscopy. At the light-microscopic level, epithelial thickness was determined histometrically. Electron micrographs were sampled at two levels of magnification, from five strata in regions of epithelial ridges and from three strata over connective tissue papillae. Standardized stereological pointcounting techniques were employed to analyze a total of 990 electron micrographs. Observations and data revealed that at the alveolar ridge the oral epithelium is truly keratinizing and comprises four strata including a 40±5 m-thick stratum corneum, which displays the oral keratin pattern. The histoand cytodifferentiation were peculiar: (1) Compared to the neighbouring gingival and hard palate epithelium, that of the alveolar crest was markedly thicker, with elongated rete ridges indicating acanthosis. (2) The cytoarchitecture was identical neither to the gingival nor to the hard palate epithelium but revealed a mixture of features typical for either of these two epithelia. Reasons for this are explained on the basis of factors, possible genetic, inherent in epithelial cells that are possibly derived from both the gingival and the palatal environment.     

Quantitative electron microscopic analysis of the epithelium of normal human alveolar mucosa

Summary The epithelium of normal human alveolar mucosa originating from the anterior vestibulum was subjected to stereologic analysis. Eight biopsies were collected half-way between the muco gingival junction and the vestibular fornix from 20 to 50 year-old females, and processed for light and electron microscopy. At two levels of magnification, electron micrographs were sampled from four artificially selected strata in regions of epithelial ridges. Stereologic point counting based on a computer-aided system for analyzing stratified epithelia served for examining a total of about 860 electron micrographs. The alveolar epithelium was 0.26 mm thick, occasionally interdigitated by short, slender connective tissue papillae, and consisted of (1) a narrow basal and suprabasal, and (2) a broad spinous and surface compartment. It displayed a differentiation pattern which, in most subjects studied, was similar to that of normal human buccal epithelium, however, on the average, produced less mature surface cells. This pattern was expressed mainly by a density increase of cytoplasmic filaments (98 Å in diameter), a concomitant decrease of the cytoplasmic ground substance, the formation of dark-cored membrane coating granules, and individually variable amounts of glycogen deposition. In some subjects, a mixed differentiation pattern was found. The structural organization of alveolar epithelium, in analogy to cheek epithelium, was compatible with the function of distensibility.

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Zusammenfassung Das Epithel der normalen menschlichen Alveolarschleimhaut im vorderen Vestibulum wurde einer stereologischen Analyse unterworfen. Acht Biopsien wurden in der Mitte zwischen mukogingivaler Grenzlinie und Fornix bei 20 bis 50 Jahre alten Frauen entnommen und für licht- und elektronenmikroskopische Studien verarbeitet. Auf zwei Vergrößerungsstufen wurden Stichproben elektronenmikroskopischer Aufnahmen aus vier Schichten im Bereich epithelialer Leisten entnommen. Insgesamt 860 Bilder wurden mit Hilfe stereologischer Punktzählverfahren analysiert. Das Alveolarepithel war im Durchschnitt 0,26 mm dick, wurde gelegentlich von kurzen, schlanken Bindegewebspapillen durchzogen und bestand aus einem schmalen basalen und suprabasalen, sowie einem breiten, homogen-strukturierten Ober-flächenkompartiment. Es wies ein Differenzierungsmuster auf, das, in der Mehrzahl der Fälle, große Ähnlichkeit mit dem des menschlichen Wangenepithels zeigte, aber durchschnittlich weniger stark ausgereifte Oberflächenzellen hervorbrachte. Dieses Muster wurde zur Hauptsache durch einen Anstieg der Filamentdichte (der Filamentdurchmesser betrug etwa 98 Å), einen entsprechenden Abfall der Volumendichte der zytoplasmatischen Grundsubstanz, der Bildung von membranversteifenden Granula und durch individuell unterschiedlich stark ausgeprägte Glykogenablagerungen geprägt. In einigen Fällen wurde ein gemischtes Differenzierungsmuster gefunden. Die strukturelle Organisation des Alveolarepithels, wie auch die des Wangenepithels, ist mit der funktionellen Dehnbarkeit dieser Schleimhautregionen vereinbar.

     

Quantitative analysis of squamous epithelium of normal palatal mucosa in guinea pigs

Summary The epithelium of intact guinea pig palate was subjected to stereologic analysis in a study of structural alterations in the keratinizing epithelium in response to wounding. Point counting procedures were employed to analyse electron micrographs sampled from three epithelial strata in biopsies collected from five animals. The differentiation pattern of the guinea pig palate epithelium displayed the following structural density gradients from basal to granular layers: descending gradients of metabolically active organelles, ascending gradient of bundled filaments coupled with the appearence of membrane coating granules and keratohyalin granules, and a plateau-like gradient of cytoplasmic ground substance. This pattern of epithelial differentiation is basically identical to that of human hard palate epithelium and epidermis. Regional and species variations in structure of keratinizing epithelia are suggested based on interepithelial differences in morphometric parameters.This investigation was supported in part by grant No. 512-4064 from the Danish State Medical Research Council and by a grant from the Calcin Foundation.The data recording and computation was performed on a guest visit at the Dental Institute, University of Zürich.     

Epithelial differentiation at the mucogingival junction: A stereological comparison of the epithelia of the vestibular gingiva and alveolar mucosa

Summary The epithelial lining of normal human vestibular gingiva and the adjoining alveolar mucosa was subjected to a comparative stereological analysis. Five biopsies collected from 11 to 12 year-old males and females were selected from a total of 14 specimens and, under standardized conditions, processed for light- and electron microscopy. At two levels of magnification, electron micrographs were sampled from five strata in the oral-gingival, and from four strata in the alveolar-mucosal epithelium, mostly in regions of epithelial ridges. Standardized sterological point counting techniques were employed to analyze a total of 710 and 540 electron micrographs from the oralgingival and the alveolar-mucosal epithelium, respectively. The two epithelia, although of similar thickness, show different differentiation patterns. The oral-gingival epithelium consists of four cytologically different strata, the major differentiation step occurring between the lower and upper stratum spinosum of epithelial ridges. Standardized stereological point counting techniques were alveolar-mucosal epithelium, consisting of two cytologically different cell compartments, displays a broad, superficial zone of differentiated flat cells, with 60% of the cytoplasm filled with a dense network of cytoplasmic filaments. The major differentiation step occurs between basal and lower spinous layers. Differentiation phenomena in both epithelia are discussed and individual variations are interpreted in view of genetically determined factors.

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Zusammenfassung Das Epithel der normalen menschlichen vestibulären Gingiva und der benachbarten Alveolarschleimhaut wurde vergleichend stereologisch analysiert. Fünf Biopsien von 11–12 Jahre alten gesunden Knaben und Mädchen, die aus insgesamt 14 Biopsien ausgewählt worden waren, wurden standardisiert für licht- und elektronenmikroskopische Studien verarbeitet. Elektronenmikroskopische Aufnahmen wurden in zwei Vergrößerungsstufen aus fünf Schichten des oralen Gingivaepithels und aus vier Schichten des Alveolarschleimhautepithels, zumeist im Bereich epithelialer Leisten, gewonnen. Insgesamt wurden 710 Bilder aus dem oralen Gingivaepithel und 540 Bilder aus dem Alveolarschleimhautepithel mit Hilfe von standardisierten stereologischen Punktzählverfahren analysiert. Die untersuchten Epithelien sind etwa gleich dick, weisen aber sehr verschiedenartige Differenzierungsmuster auf. Das orale Gingivaepithel besteht aus vier zytologisch unterschiedlichen Schichten und bildet ein parakeratinisiertes, 0,1 mm dickes Stratum corneum, wobei der Hauptdifferenzierungsschritt zwischen dem unteren und dem oberen Stratum spinosum im Bereich der epithelialen Leisten erfolgt. Das Alveolarschleimhautepithel weist zwei zytologisch unterschiedliche Zellkompartimente auf und bildet eine breite oberflächliche Lage flacher und differenzierter Zellen, deren Zytoplasma zu 60% aus einem dichten Maschenwerk zytoplasmatischer Filamente besteht. Der Hauptdifferenzierungsschritt dieses Epithels liegt zwischen dem Stratum basale und dem unteren Stratum spinosum. Die verschiedenen Differenzierungsvorgänge werden diskutiert und individuelle Variationen, die in beiden Epithelien auftreten, im Hinblick auf genetische Faktoren erklärt.

     

Ultrastructural study of the precursor to fungiform papillae prior to the arrival of sensory nerves in the fetal rat.

The structure of precursors to fungiform papillae without taste buds, prior to the arrival of sensory nerve fibers at the papillae, was examined in the fetal rat on embryonic day 13 (E13) and 16 (E16) by light and transmission electron microscopy in an attempt to clarify the mechanism of morphogenesis of these papillae. At E13, a row of rudiments of fungiform papillae was arranged along both sides of the median sulcus of the lingual dorsal surface, and each row consisted of about 10 rudiments. There was no apparent direct contact between papillae rudiments and sensory nerves at this time. Bilaterally towards the lateral side of the tongue, adjacent to these first rudiments of fungiform papillae, a series of cord-like invaginations of the dorsal epithelium of the tongue into the underlying connective tissue, representing additional papillary primordia parallel to the first row, was observed. The basal end of each invagination was enlarged as a round bulge, indented at its tip by a mound of fibroblasts protruding into the bulge. At E16 there was still no apparent direct contact between rudiments of fungiform papillae and sensory nerves. Each rudiment apically contained a spherical core of aggregating cells, which consisted of a dense assembly of large, oval cells unlike those in other areas of the lingual dorsal epithelium. The differentiation of these aggregated cells was unclear. The basal lamina was clearly recognizable between the epithelium of the rudiment of fungiform papillae and the underlying connective tissue. Spherical structures, which appeared to be sections of the cord-like invaginations of the lingual epithelium that appeared on E13, were observed within the connective tissue separated from the dorsal lingual epithelium. Transverse sections of such structures revealed four concentric layers of cells: a central core, an inner shell, an outer shell, and a layer of large cells. Bundles of fibers were arranged in the central core, and the diameters of bundles varied somewhat depending on the depth of the primordia within the connective tissue and their distance from the median sulcus. Ultrastructural features of cells in the outer shell differed significantly in rudiments close to the lingual epithelium as compared to those in deeper areas of connective tissue. Around the outer shell there was a large-cell layer consisting of one to three layers of radially elongated, oval cells that contained many variously sized, electron-dense, round granules. Large numbers of fibroblasts formed dense aggregates around each spherical rudiment, and were separated by the basal lamina from the large-cell epithelial layer. Progressing from deep-lying levels of the rudiments of the papillae to levels close to the lingual surface epithelium, the central core, inner shell, and outer shell gradually disappeared from the invaginated papillary cords.     

The surface structure of the completely and incompletely orthokeratinized oral epithelium in the rat: a light, scanning and transmission electron microscope study.

Mucosa from the hard and soft palates, molar gingiva, cheek and dorsal surface of the tongue of the rat was examined in the light microscope, following Mallory's triple connective tissue stain, and in the scanning and transmission electron microscopes. The epithelium covering the hard palate, gingiva, the smooth band of mucosa at the junction of the hard and soft palates, intermediate zones of the soft palate, fungiform papilla-like structures in the central zone of the soft palate, the fungiform papillae, and the more superficial part and posterior surfaces of the filiform papillae of the tongue all exhibited complete orthokeratinization. The oral surfaces of the epithelial cells in all these areas had a honeycomb pattern of interconnecting ridges surrounding depressions. Imprints of the overlying cells that had been desquamated were apparent, and the lateral boundaries between the cells were formed by two raised ridges separated by a gap. The epithelium covering the cheek, central zone of the soft palate apart from the fungiform papilla-like structures, lateral zones of the soft palate, gingival crevice, and the mucosa between the fungiform and filiform papillae of the tongue all exhibited incomplete orthokeratinization. The oral surfaces of the epithelial cells in all these areas were relatively smooth and did not exhibit a honeycomb pattern of interconnecting ridges. Imprints of the overlying cells that had been desquamated and the lateral boundaries between the cells were only very occasionally found. In the transmission electron microscope the outlines of the cells were compatible with the surface patterns seen in the scanning electron microscope. The possible relationships between the degree of orthokeratinization and ultrastructure of the various epithelia are discussed.     

Scanning electron microscopy of isolated epithelium of the murine gastrointestinal tract: morphology of the basal surface and evidence for paracrinelike cells

By using the method of Bjerknes and Cheng, isolated murine gastrointestinal epithelial sheets were prepared for scanning electron microscopy. Examination of isolated epithelium from fundic stomach revealed numerous branched gastric glands. Parietal cells were easily detected bulging from the basal surface of the glandular epithelium. The basal surface membrane of parietal cells appeared smooth, with only sparse microvilluslike projections, whereas adjacent glandular cells had numerous 1- to 2-micron fingerlike projections which interdigitated laterally with similar processes from adjacent cells. Occasionally, paracrinelike cells having long cytoplasmic processes ranging from 10 to 20 micron in length were observed on the basal epithelial surface of the stomach and the colon, but not the small intestine. In isolated intestinal epithelia, the basal surface of crypt epithelial cells showed extensive cytoplasmic interdigitations, but no distinct morphology permitting recognition of individual cell types. Various stages of intestinal crypt bifurcation were seen. Craterlike spaces in the basal surface of crypt epithelium, presumably due to migrating leukocytes, were also numerous. Examination of the luminal surface of the isolated intestinal epithelium revealed that intimate associations between epithelium and mucosal-associated microorganisms were maintained, thus suggesting that minimal alterations in surface morphology were incurred by epithelial isolation. These observations on epithelial structure suggest that isolated gastrointestinal epithelia may be well suited for physiological studies of epithelial function and interactions with the microbial flora.     

Quantitative analysis of epithelial changes during wound healing in palatal mucosa of guinea pigs

Summary The epithelium of wounded guinea pig palate was subjected to stereologic analysis. A total of 18 biopsies (animals) were used. Biopsies were taken at 18, 48, 96 and 120 h after wounding. Point counting procedures were employed to analyse electron micrographs sampled from one (18 h) or two epithelial strata (48, 96 and 120 h). The essential modulations in epithelial structure as wound healing proceeds were as follows: During the early phases characterized by formation and advancement of epithelial lips (18 and 48 h), migrating cells converged towards a cell type which structurally was less differentiated than normal basal cells. This alteration was expressed by a decrease in volume density of cytoplasmic organelles, mainly mitochondria, free ribosomes and tonofilament bundles, coupled with an increase in volume density of lysosomal bodies. Concomitantly, the volume density of cytoplasmic ground substance rose markedly. Subsequent to fusion of contralateral migratory lips (96 and 120 h) reversion to normal epithelial structure was indicated by the increment in magnitude of basal cell parameters. Further structural density gradients from basal towards upper cell layers appeared. This pattern was mainly displayed by mitochondria, free ribosomes, and tonofilament bundles. The magnitude and gradation of most tissue and cell parameters were not yet re-established at 120 h. The density of tonofilament bundles and the density level of cytoplasmic ground substance in particular deviated.This investigation was supported in part by grants No. 512-5958 and No. 512-5151 from the Danish State Medical Research Council     

Light and electron microscopic studies on the development of the spiral prominence of the cochlear duct of the fetal guinea pig

The anlage of the spiral prominence can be seen on the 37th day of development as a small protrusion of the epithelium towards the lumen of the cochlear duct. During the further progress, the spiral prominence more distinctly protrudes by augmentation of the vascularized connective tissue. In the epithelial cells pinocytotic vesicles near the plasmalemma are seen earliest lateral and basal on the 37th day, apical on the 39th day. The epithelial cells send basal cytoplasmic extensions towards the connective tissue. Starting on the 44th day, small invaginations of connective tissue extend into the epithelium, remaining separated from the epithelial cells by the basal lamina. Until the 48th day, the monostratified epithelium remains columnar, thereafter it changes to cuboidal or flat. Towards the end of the development, the invaginations of the connective tissue nearly reach the surface of the epithelium, being separated from the endolymph by a small epithelial area.     

Response of basal epithelial cell surface and Cytoskeleton to solubilized extracellular matrix molecules

Corneal epithelium removed from underlying extracellular matrix (ECM) extends numerous cytoplasmic processes (blebs) from the formerly smooth basal surface. If blebbing epithelia are grown on collagen gels or lens capsules in vitro, the basal surface flattens and takes on the smooth contour typical of epithelium in contact with basal lamina in situ. This study examines the effect of soluble extracellular matrix components on the basal surface. Corneal epithelia from 9- to 11-d-old chick embryos were isolated with trypsin-collagenase or ethylenediamine tetraacetic acid, then placed on Millipore filters (Millipore Corp., Bedford, Mass.), and cultured at the medium-air interface. Media were prepared with no serum, with 10% of calf serum, or with serum from which plasma fibronectin was removed. Epithelia grown on filters in this medium continue to bleb for the duration of the experiments (12-14 h). If soluble collagen, laminin, or fibronectin is added to the medium, however, blebs are withdrawn and by 2-6 h the basal surface is flat. Epithelia grown on filters in the presence of albumin, IgG, or glycosaminoglycans continue to bleb. Epithelia cultured on solid substrata, such as glass, also continue to bleb if ECM is absent from the medium. The basal cell cortex in situ contains a compact cortical mat of filaments that decorate with S-1 myosin subfragments; some, if not all, of these filaments point away from the plasmalemma. The actin filaments disperse into the cytoplasmic processes during blebbing and now many appear to point toward the plasmalemma. In isolated epithelia that flatten in response to soluble collagens, laminin, and fibronectin, the actin filaments reform the basal cortical mat typical or epithelial in situ. Thus, extracellular macromolecules influence and organize not only the basal cell surface but also the actin-rich basal cell cortex of epithelial cells.     

Stereo architecture of the interface of the epithelial cell layer and connective tissue core of the foliate papilla in the rabbit tongue.

The three-dimensional relationship between the epithelial cell layer and the underlying connective tissue core (CTC) of the foliate papilla of the rabbit tongue was studied by scanning electron microscopy after removal of the epithelial cell layer. The foliate papillae were fixed in Karnovsky's fixative, and the epithelial cell layers were exposed to long-term hydrochloric acid treatment (3.5 N HCl for 2-3 weeks at room temperature). The foliate papillae consisted of ridges and grooves located on the posterolateral margin of the tongue. They appeared as linear projections or ridges of lingual mucosa roughly perpendicular to the longitudinal axis of the tongue. These projections or ridges were parallel to one another and separated by grooves. After removal of the epithelium, two kinds of CTC folds appeared: one was the septal fold of CTC which runs in the central portion under each linear projection or ridge, and the other consisted of two sheets of groove side folds of CTC which run along both sides of the former and face the groove side epithelium. It was revealed that there are two sheets of septal epithelial processes, and each of them fits in between each septal fold and groove side fold of CTC. Numerous taste buds were located in the groove side epithelia, and their pores faced the surface of the groove. On the hollow surfaces that appeared on the surface of the groove side fold of CTC after removal of the epithelial cells with taste buds, nerve-terminal-like structures were encountered. Some openings of the ducts of small lingual glands were arranged linearly on the underside of the basal portion of each groove side epithelium.     

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.     

Asymmetric stem-cell divisions define the architecture of human oesophageal epithelium

In spite of its clinical importance, little is known about the stem-cell compartment of the human oesophageal epithelium [1,2]. The epithelial basal layer consists of two distinct zones, one overlying the papillae of the supporting connective tissue (PBL) and the other covering the interpapillary zone (IBL) [3]. In examining the oesophageal basal layer, we found that proliferating cells were rare in the IBL and a high proportion of mitoses were asymmetrical, giving rise to one basal daughter and one suprabasal, differentiating daughter. In the PBL, mitoses were more frequent and predominantly symmetrical. The IBL was characterised by low expression of ?1 integrins and high expression of the beta2 laminin chain. By combining fluorescence-activated cell sorting (FACS) with in vitro clonal analysis, we obtained evidence that the IBL is enriched for stem cells. A normal oesophageal epithelium with asymmetric divisions was reconstituted on denuded oesophageal connective tissue. In contrast, asymmetric divisions were not sustained on skin connective tissue, and the epithelium formed resembled epidermis. We propose that stem cells located in the IBL give rise to differentiating daughters through asymmetric divisions in response to cues from the underlying basement membrane. Until now, stem-cell fate in stratified squamous epithelia was believed to be achieved largely through populational asymmetry [4-6].     

Connective tissue influences on patterns of epithelial architecture and keratinization in skin and oral mucosa of the adult mouse

Summary Epithelial-mesenchymal interactions play an important role during embryogenesis but it is uncertain whether such interactions influence the maintenance of epithelial structure in the adult. To examine this problem, separated epithelial and connective tissue components of skin and mucosae from various regions of adult mice were homoor heterotypically recombined and transplanted to histocompatible hosts. The patterns of tissue architecture and keratinization of the resultant epithelia were examined for changes indicative of mesenchymal influences on the epithelial phenotype. Each type of epithelium, in some recombinations, fully conserved its normal pattern of phenotypic expression indicating that subepithelial connective tissue from all regions is permissive and that regionally-specific connective tissue influences are not necessary for conservation of epithelial specificity. In other recombinations, however, the epithelium acquired features of tissue architecture or keratinization typical of the epithelium normally associated with the connective tissue component, indicating directive influ ences from the connective tissue. The patterns of epithelial response observed suggest that there may be separate connective tissue influences on epithelial architecture and cytodifferentiation and that there is a regionally-related variation in the competence of epithelia to respond to these influences.Supported by NIH NIDR 2 R01 DE05190     

Keratin filaments of epithelial and taste-bud cells in the circumvallate papillae of adult and developing mice

Summary Keratin filaments of epithelial- and taste-bud cells in the circumvallate papillae of adult and developing mice were studied by immunocytochemistry using monoclonal antikeratin antibodies (PKK2 and PKK3) and by conventional electron microscopy. Elongated cells (type-I,-II, and-III cells) of the taste buds were stained by PKK3 antibody, which reacts with 45-kdalton keratin, whereas basal cells of the taste buds and surrounding epithelial cells showed negative staining with PKK3. Such PKK3-reactive cells occurred at 0 day after birth, when taste-buds first appeared in the dorsal surface epithelium of the papillae. Thus 45-kdalton keratin seems to be an excellent immunocytochemical marker for identifying taste-bud cells. Epithelial cells in all layers of the trench wall and basal layer cells of the dorsal surface contained densely aggregated bundles of keratin filaments that reacted with PKK2 antibody, but not with PKK3. In contrast, taste-bud cells and spinous and granular layer cells of the dorsal surface possessed loose aggregated bundles of filaments that reacted with PKK3, but not with PKK2. These results suggest that the aggregation and distribution pattern of keratin filaments may reflect differences in the keratin subtypes that comprise these filaments.     

Cytokeratin expression in human tongue epithelium.

The epithelium of the human tongue shows diverse morphological variations from one site to another and even within the epithelium of the same papilla. This complexity has led to confusion regarding tongue epithelium as being orthokeratinized, parakeratinized, or nonkeratinized. Cytokeratins have been shown to characterize different epithelia. The present paper describes cytokeratin expression by adult tongue epithelia and relates their distribution to morphology. Six healthy human tongue specimens were obtained after plastic surgery and cytokeratin expression was investigated immunohistochemically, using a panel of 15 antibodies for cytoskeletal proteins, and biochemically using two-dimensional gel electrophoresis. The results showed that the ventral and lateral surfaces of the tongue are related to the nonkeratinizing stratified squamous epithelia, esophageal type, whereas the dorsal surface showed mixed expression of cytokeratins. In the tip of filiform and on the surface of fungiform papillae, cytokeratins of terminal differentiation are expressed as skin type; and in the rest of the papillae as well as in interpapillary areas, the epithelium expresses esophageal type cytokeratins. Certain simple epithelial cytokeratins were found in taste buds. Cytokeratin 19 was also detected in the basal cell layer of all esophageal type epithelia in the tongue. The present results provide basis for studies on the biological events in epithelial differentiation during development and in pathology.     

Ultrastructural organization of the hamster renal pelvis.

The renal pelvis of the hamster has been studied by light microscopy (epoxy resin sections), transmission electron microscopy, and morphometric analysis of electron micrographs. Three morphologically distinct epithelia line the pelvis, and each covers a different zone of the kidney. A thin epithelium covering the outer medulla (OM) consists of two cell types: (1) granular cells are most numerous and have apically positioned granules which stain intensely with toluidine blue, are membrane-bound, and contain a fine particulate matter that stains light grey to black in electron micrographs. (2) Basal cells do not have granules, are confined to the basal lamina region, and do not reach the mucosal epithelial surface. The inner medulla (IM) is covered by a pelvic epithelium morphologically similar to collecting duct epithelium of IM. Some cells in this portion of the pelvic epithelium (IM) stain intensely dark with toluidine blue, osmium tetroxide, lead, and uranyl acetate. Transitional epithelium, which separates cortex (C) from pelvic urine, has an asymmetric luminal plasma membrane and discoid vesicles, each of which is similar to those previously observed in mammalian ureter and urinary bladder epithelia. Based on morphological comparisons with other epithelia, the IM and OM pelvic epithelia would appear permeable to solutes and/or water, while the transitional epithelium covering the C appears relatively impermeable. It would also appear that the exchange of solutes and water between pelvic urine and OM would involve capillaries, primarily, since morphometric analysis showed that both fenestrated and continuous capillaries of the OM were extremely abundant (greater than 60% of OM pelvic surface area) just under the thin pelvic epithelium.     

Fine-structural observations on the gill filaments of the fresh-water crayfish, Astacus pallipes Lereboullet

The gills of the crayfish are described with special reference to the tissues of the branchial filaments. An account is given of a possible pathway of blood flow through the filaments to ensure maximum oxygenation at the epithelial surface. The branchial epithelium consists of a thin, subcuticular layer with an indented apical plasma membrane, and projecting cell bodies. The basal epithelial surface adjacent to the haemolymph possesses the deep membrane infoldings characteristic of resorptive epithelia. Ion and water movement across the epithelium is discussed.     

The influence of subepithelial connective tissues on epithelial proliferation in the adult mouse

Summary Subepithelial connective tissue is capable of modulating the pattern of histodifferentiation of stratified epithelia from adult animals, but it is not known whether the supporting connective tissue also influences epithelial proliferative activity. Epithelial and connective tissues of murine skin and oral mucosa, differing in their morphology and proliferative activity, were separated and heterotypically recombined prior to grafting to histocompatible hosts. After 3 or 8 weeks in situ, mitotic activity was determined following the administration of vinblastine sulfate. Although the mitotic activity in each of the epithelia could be modulated by some connective tissues, there was no distinct pattern of behavior. In combination with connective tissues from tongue or palate, the ear epidermis acquired a significantly increased mitotic activity. In contrast, when oral epithelia with high mitotic activity were recombined with dermal connective tissue, there was usually a significant reduction in proliferative activity. As there was no apparent association between mitotic activity and the induced changes in either organization or histodifferentiation, it is suggested that subepithelial connective tissue is capable of directly influencing the mitotic activity in the overlying epithelium.