Apoptosis of the reduced enamel epithelium and its implications for bone resorption during tooth eruption

Bone remodeling, the selective deposition and resorption of bone, is an important cause of tooth eruption. During tooth eruption, reduced enamel epithelia of the enamel organ interact with follicle cells to recruit osteoclasts for bone remodeling. However, little is known about the relationship between cellular activity of reduced enamel epithelium and bone resorption during tooth eruption. The purpose of this study was to investigate the effect of apoptosis in reduced enamel epithelium on osteoclastogenesis and its implications for bone resorption. We have analyzed erupting mandibular molars in mice by TdT-mediated dUTP-biotin nick end labeling assay, tartrate-resistant acid phosphatase (TRAP) staining, and immunohistochemistry. TRAP-positive cells were detected in the osteoclasts near both the buccal and lingual sides of tooth socket at postnatal day 0 (PN0). They significantly increased until PN3 and decreased thereafter as the tooth erupted. Interestingly, apoptosis was barely detected in the reduced enamel epithelium at PN3 but clearly at PN7. A few apoptotic cells were also investigated within the dental follicle surrounding developing tooth at PN7 and PN10. We observed apoptotic osteoblast-lineage cells along the inner margin of alveolar bone facing the buccal cusp and at the base of the bony crypt at PN3 decreasing until PN10. In contrast, expression levels of bone sialoprotein increased at PN10 compared to levels at PN3. These results suggest that apoptosis of reduced enamel epithelium resulted in a reduction of osteoclast activity and of bone resorption mediated by dental follicle during tooth eruption.     

The role of epithelial remodelling in tooth eruption in larval zebrafish

Based on light and transmission electron-microscopic observations on erupting first-generation teeth in the zebrafish, Danio rerio, we propose a biphasic mechanism for tooth eruption: (1). formation of an epithelial crypt prior to eruption of the tooth, possibly as a result of constraints in the epithelium resulting from the growth of adjacent tooth germs, and (2). detachment of cellular interdigitations both within the pharyngeal epithelium, at the pharyngeal epithelium/enamel organ boundary, and between the outer and inner dental epithelium, resulting in the exposure of the tooth tip in the crypt, immediately after tooth ankylosis. Later, further detachment of interdigitations between the inner and the outer enamel epithelium unfolds the epithelium even more and leads to a more pronounced exposure of the tooth tip. The presence of small patches of non-collagenous matrix on the outer surface of the tooth close to where it merges with the attachment bone is interpreted as a device to prevent complete detachment of the enamel organ. The biphasic nature of the mechanism for tooth eruption is supported by observations on in vitro cultured heads. First-generation teeth develop normally and crypts are formed, as under in vivo conditions, but the teeth fail to erupt. Taken together, our observations suggest that epithelial remodelling plays a crucial role in eruption of the teeth in this model organism.     

INNER ENAMEL EPITHELIUM CELL PRODUCTION RATES

Crowning of all four rat incisors slows down their eruption rate, leading to a diminution of cell production by the inner enamel epithelium (IEE). The production rate by 100 progenitor cells is dominated as production rate %. The normal IEE in rats weighing 200 g produces daily 67 cells %. In the crowned incisor this rate drops to 15 cells % while during accelerated eruption the IEE produces daily 152 cells %. In all three conditions the ratio between eruption rate and IEE cell production rate remains unchanged and an eruption of 6.4 μm reflects a production of 1 cell % by the IEE. A method which relates labelled cell displacement along the IEE with its cell production rate is described and its applicability to other cell renewal systems discussed.     

Fate map of the dental mesenchyme: dynamic development of the dental papilla and follicle

At the bud stage of tooth development the neural crest derived mesenchyme condenses around the dental epithelium. As the tooth germ develops and proceeds to the cap stage, the epithelial cervical loops grow and appear to wrap around the condensed mesenchyme, enclosing the cells of the forming dental papilla. We have fate mapped the dental mesenchyme, using in vitro tissue culture combined with vital cell labelling and tissue grafting, and show that the dental mesenchyme is a much more dynamic population then previously suggested. At the bud stage the mesenchymal cells adjacent to the tip of the bud form both the dental papilla and dental follicle. At the early cap stage a small population of highly proliferative mesenchymal cells in close proximity to the inner dental epithelium and primary enamel knot provide the major contribution to the dental papilla. These cells are located between the cervical loops, within a region we have called the body of the enamel organ, and proliferate in concert with the epithelium to create the dental papilla. The condensed dental mesenchymal cells that are not located between the body of the enamel organ, and therefore are at a distance from the primary enamel knot, contribute to the dental follicle, and also the apical part of the papilla, where the roots will ultimately develop. Some cells in the presumptive dental papilla at the cap stage contribute to the follicle at the bell stage, indicating that the dental papilla and dental follicle are still not defined populations at this stage. These lineage-tracing experiments highlight the difficulty of targeting the papilla and presumptive odontoblasts at early stages of tooth development. We show that at the cap stage, cells destined to form the follicle are still competent to form dental papilla specific cell types, such as odontoblasts, and produce dentin, if placed in contact with the inner dental epithelium. Cell fate of the dental mesenchyme at this stage is therefore determined by the epithelium.     

The primary enamel knot determines the position of the first buccal cusp in developing mice molars

The enamel knot (EK), which is located in the center of bud and cap stage tooth germs, is a transitory cluster of non-dividing epithelial cells. The EK acts as a signaling center that provides positional information for tooth morphogenesis and regulates the growth of tooth cusps by inducing secondary EKs. The morphological, cellular, and molecular events leading to the relationship between the primary and secondary EKs have not been described clearly. This study investigated the relationship between the primary and secondary EKs in the maxillary and mandibular first molars of mice. The location of the primary EK and secondary EKs was investigated by chasing Fgf4 expression patterns in tooth germ at some intervals of in vitro culture, and the relationship between the primary EK and secondary EK was examined by tracing the primary EK cells in the E13.5 tooth germs which were frontally half sliced to expose the primary EK. After 48 hr, the primary EK cells in the sliced tooth germs were located on the buccal secondary EKs, which correspond to the future paracone in maxilla and protoconid in mandible. The Bmp4 expression in buccal part of the dental mesenchyme might be related with the lower growth in buccal epithelium than in lingual epithelium, and the Msx2 expressing area in epithelium was overlapped with the enamel cord (or septum) and cell dense area. The enamel cord might connect the primary EK with enamel navel to fix the location of the primary EK in the buccal side during the cap to bell stages. Overall, these results suggest that primary EK cells strictly contribute to form the paracone or protoconid, which are the main cusps of the tooth in the maxilla or mandible.     

Fluorescence histochemical observations on catecholamine-containing cell bodies in Auerbach's plexus

Summary The nature and distribution of cell contacts have been examined in the human enamel organ in bell stage. The lateral cell surfaces of secretory ameloblasts are linked at their distal (apical) and proximal (basal) parts by junctional complexes consisting of tight junctions, large intermediate junctions (zonulae adherentes), occasional gap junctions and one or more series of desmosomes. Scattered desmosomes and large gap junctions link epithelial cells of the external enamel epithelium, stellate reticulum, stratum intermedium and internal enamel epithelium including secretory ameloblasts. Furthermore the above-mentioned layers are also linked together by desmosomes and gap junctions.With increasing maturation of the enamel organ an increase in size and number of gap junctions is observed. Some possible implications of the role of the different junctions are considered. The gap junctions probably participate in cell differentiation in the normal morphogenesis of the teeth as well as in metabolic and ionic coupling of the cells of the enamel organ. By means of tight junctions, adjacent secretory ameloblasts cooperate to form a physical barrier which might prevent the diffusion of some types of molecules or substances (e.g. secretory material distally and acid mucopolysaccharides proximally) through the interspaces between the cells. Adhering junctions might assist in regulation of the mechanical properties of the enamel organ as a whole.This work was supported by grants from Statens almindelige Videnskabsfond, Copenhagen, and the Association for the Aid of the Crippled Children, New York.     

Aspects of cell proliferation kinetics of the inner dental epithelium during mouse molar and incisor morphogenesis: a reappraisal of the role of the enamel knot area.

First lower E-14 and E-16 mouse molars and E-13 lower incisors were cultured in vitro and either sequentially or continuously labelled with BrdU (5-bromo-2'-deoxyuridine). The behaviour of the non-cycling inner dental epithelial cells emerging from the enamel knot area of the molars was analysed by 3D (three dimensional) reconstructions of serial sections. These cells, as well as slow cycling cells underwent a coordinated temporo-spatial patterning leading to their patchy segregation at the tips of the forming cusps. In incisors (in vitro and in vivo), non-cycling cells were also present in the inner dental epithelium of the enamel knot area. However, these cells were not redistributed during incisor morphogenesis. These non-dividing inner dental epithelium cells of the enamel knot area which are either redistributed or not according to the tooth type specific morphogenesis might represent the organizers of morphogenetic units (OMU), the cusps.     

Histological and three dimensional organization of the odontogenic organ in the lower incisor of 100 gram rats.

A three dimensional reconstruction of the epithelial tissue at the apical end of the lower rat incisor was made from serial 1 mum thick cross sections. This tissue formed an elongated structure, called the odontogenic organ, which was composed of a bulbous and a "U"-shaped part. Both parts were joined to one another at the posterior aspect of the apical foramen. The bulbous part of the odontogenic organ was situated at the lingual side of the "U"-shaped part and protruded anteriorly over the pulp. It was formed by cells of the outer dental epithelium and stellate reticulum whose organization suggested that the bulbous part was important in the production of cells for renewal of all the epithelia of the incisor. The "U"-shaped part of the odontogenic organ was apparently derived from the bulbous part and delineated the pulp by forming the lateral, mesial and labial sidewalls around the apical foramen. It was composed of all the epithelial cell types recognizable as precursors to (a) cells of the enamel organ which form the enamel, and (b) Hertwig's epithelial root sheath, a part of the odontogenic organ which induces the formation of dentin on the lingual aspect of the incisor.     

Observations on the cervix uteri of the squirrel monkey

The squirrel monkey uterine cervix was studied macroscopically and microscopically in intact and ovariectomized monkeys. The effect in ovariectomized monkeys of estradiol dipropionate or progesterone of both given after estrogen priming was studied by PAS staining. The lower portion of the cervix was dilated to form a vestibule into which projected fibromuscular colliculi which arose from the isthmic end of the cervix. The stratified squamous epithelium of the vagina was continuous through the external os with a similar epithelium lining the vestibule and covering the external surfaces of the colliculi. The transitional zone between the stratified epithelium and columnar cells was variable. The colliculi were covered internally with mucosal folds of columnar epithelium continuous with those of the endocervical canal. Glycogen concentration in the smooth muscle did not fluctuate markedly, irrespective of the hormones used. Glycogen granules were more numerous in the stratified squamous epithelium. Malt-diastase-resistant material appeared to be more abundant in the columnar epithelium and glandular lumina when the monkeys received both hormones than when they received solely estrogen or progesterone.     

The development and differentiation of the parabronchial unit in quail (Coturnix coturnix)

The present study has been inspired by the conflicting data in the relevant literature concerning the embryogenesis of cell types of the parabronchial epithelium and the formation, discharge and distribution of trilaminar substance and lamellar bodies. Lung tissue from embryonic, newly hatched, immature and mature quail was subjected to standard processing for light and transmission electron microscopy. The parabronchial rudiments form shallow primitive atria on embryonic day 13. The precursors of granular cells differentiate with lamellar bodies in their cytoplasm. The residual population of non-granular epithelial cells is the common source for the differentiation of primitive squamous atrial and respiratory cells, the potential producers of trilaminar substance. The primitive squamous atrial cells sprout as branching infundibular canaliculi into the mesenchyme on embryonic day 14. The infundibular epithelium differentiates into the squamous respiratory cells that constitute with blood capillaries the blood-air barrier. Not until the time of hatching could the trilaminar substance be visualized being produced by squamous atrial and respiratory cells. In the late prehatching and early posthatching period the granular cells intensely escalate the production and discharge of lamellar bodies. The lamellar bodies form, together with sheets of trilaminar substance, mixed multilayered masses in atria. They disappear fast in the successive posthatching period. The formation of trilaminar substance in squamous atrial and respiratory cells is governed by the agranular endoplasmic reticulum, the cisternae of which take part in the formation of trilaminar units. The gas exchange tissue is predominantly represented by infundibula in immature quail. The posthatching growth of the gas exchange tissue of immature to mature quail occurs via intense multiplication of air and blood capillaries.     

The effect of rate of eruption on periodontal ligament glycosylaminoglycan content and enamel formation in the rat incisor

The rate of eruption of rat mandibular incisors was either increased by cutting one tooth out of occlusion or eliminated by means of pinning. The effects of such changes in eruption rate on the sulphated glycosylaminoglycan content of the periodontal ligaments was analysed. The length of the enamel secretory zone and the composition of the developing enamel matrix protein was also compared. Sulphated glycosylaminoglycan content of the periodontal ligament increased fourfold (P<0.001) during accelerated eruption but decreased to a corresponding extent (P<0.001) in the absence of eruption, when compared with controls. The length of the enamel secretory zone was also significantly reduced in the immobilised teeth, although the protein content was similar compared with controls. The results demonstrate the differential response to varied eruption rates of the periodontal ligament and enamel, particularly in respect of the extracellular matrix. The data are consistent with the view that the ground substance of the periodontal ligament plays a role in the generation of the eruptive force.     

Unicuspid and bicuspid tooth crown formation in squamates

The molecular and developmental factors that regulate tooth morphogenesis in nonmammalian species, such as snakes and lizards, have received relatively little attention compared to mammals. Here we describe the development of unicuspid and bicuspid teeth in squamate species. The simple, cone-shaped tooth crown of the bearded dragon and ball python is established at cap stage and fixed in shape by the differentiation of cells and the secretion of dental matrices. Enamel production, as demonstrated by amelogenin expression, occurs relatively earlier in squamate teeth than in mouse molars. We suggest that the early differentiation in squamate unicuspid teeth at cap stage correlates with a more rudimentary tooth crown shape. The leopard gecko can form a bicuspid tooth crown despite the early onset of differentiation. Cusp formation in the gecko does not occur by the folding of the inner enamel epithelium, as in the mouse molar, but by the differential secretion of enamel. Ameloblasts forming the enamel epithelial bulge, a central swelling of cells in the inner enamel epithelium, secrete amelogenin at cap stage, but cease to do so by bell stage. Meanwhile, other ameloblasts in the inner enamel epithelium continue to secrete enamel, forming cusp tips on either side of the bulge. Bulge cells specifically express the gene Bmp2, which we suggest serves as a pro-differentiation signal for cells of the gecko enamel organ. In this regard, the enamel epithelial bulge of the gecko may be more functionally analogous to the secondary enamel knot of mammals than the primary enamel knot.     

Expression patterns of the homeobox gene, Hox-8, in the mouse embryo suggest a role in specifying tooth initiation and shape.

We have studied the expression patterns of the newly isolated homeobox gene, Hox-8 by in situ hybridisation to sections of the developing heads of mouse embryos between E9 and E17.5, and compared them to Hox-7 expression patterns in adjacent sections. This paper concentrates on the interesting expression patterns of Hox-8 during initiation and development of the molar and incisor teeth. Hox-8 expression domains are present in the neural crest-derived mesenchyme beneath sites of future tooth formation, in a proximo-distal gradient. Tooth development is initiated in the oral epithelium which subsequently thickens in discrete sites and invaginates to form the dental lamina. Hox-8 expression in mouse oral epithelium is first evident at the sites of the dental placodes, suggesting a role in the specification of tooth position. Subsequently, in molar teeth, this patch of Hox-8 expressing epithelium becomes incorporated within the buccal aspect of the invaginating dental lamina to form part of the external enamel epithelium of the cap stage tooth germ. This locus of Hox-8 expression becomes continuous with new sites of Hox-8 expression in the enamel navel, septum, knot and internal enamel epithelium. The transitory enamel knot, septum and navel were postulated, long ago, to be involved in specifying tooth shape, causing the inflection of the first buccal cusp, but this theory has been largely ignored. Interestingly, in the conical incisor teeth, the enamel navel, septum and knot are absent, and Hox-8 has a symmetrical expression pattern. Our demonstration of the precise expression patterns of Hox-8 in the early dental placodes and their subsequent association with the enamel knot, septum and navel provide the first molecular clues to the basis of patterning in the dentition and the association of tooth position with tooth shape: an association all the more intriguing in view of the evolutionary robustness of the patterning mechanism, and the known role of homeobox genes in Drosophila pattern formation. At the bell stage of tooth development, Hox-8 expression switches tissue layers, being absent from the differentiating epithelial ameloblasts and turned on in the differentiating mesenchymal odontoblasts. Hox-7 is expressed in the mesenchyme of the dental papilla and follicle at all stages. This reciprocity of expression suggests an interactive role between Hox-7, Hox-8 and other genes in regulating epithelial mesenchymal interactions during dental differentiation.(ABSTRACT TRUNCATED AT 400 WORDS)     

Immunocytochemical and biochemical detection of EMMPRIN in the rat tooth germ: differentiation-dependent co-expression with MMPs and co-localization with caveolin-1 in membrane rafts of dental epithelial cells

In tooth development matrix metalloproteinases (MMPs) are under the control of several regulatory mechanisms including the upregulation of expression by inducers and downregulation by inhibitors. The aim of the present study was to monitor the occurrence and distribution pattern of the extracellular matrix metalloproteinase inducer (EMMPRIN), the metalloproteinases MMP-2 and MT1-MMP and caveolin-1 during the cap and bell stage of rat molar tooth germs by means of immunocytochemistry. Strong EMMPRIN immunoreactivity was detected on the cell membranes of ameloblasts and cells of the stratum intermedium in the bell stage of the enamel organ. Differentiating odontoblasts exhibited intense EMMPRIN immunoreactivity, especially at their distal ends. Caveolin-1 immunoreactivity was evident in cells of the internal enamel epithelium and in ameloblasts. Double immunofluorescence studies revealed a focal co-localization between caveolin-1 and EMMPRIN in ameloblastic cells. Finally, western blotting experiments demonstrated the expression of EMMPRIN and caveolin-1 in dental epithelial cells (HAT-7 cells). A substantial part of EMMPRIN was detected in the detergent-insoluble caveolin-1-containing low-density raft membrane fraction of HAT-7 cells suggesting a partial localization within lipid rafts. The differentiation-dependent co-expression of MMPs with EMMPRIN in the enamel organ and in odontoblasts indicates that EMMPRIN takes part in the induction of proteolytic enzymes in the rat tooth germ. The localization of EMMPRIN in membrane rafts provides a basis for further investigations on the role of caveolin-1 in EMMPRIN-mediated signal transduction cascades in ameloblasts.     

Structural features of forming and developing blood capillaries of the enamel organ of rat molar tooth germs observed by light and electron microscopy

The process of vascularization of the enamel organ, a unique epithelial structure, occurs when the tooth germ is fully developed, i.e., at the onset of dentinogenesis. Although the three-dimensional organization of the capillaries has been previously investigated, the structural features underlying the formation of the new capillaries remains poorly understood. Thus, in the hope of better understanding the mechanism of formation of the stellate reticulum capillaries, upper first molar tooth germs of newborn and 3-day-old rats were fixed in glutaraldehyde-formaldehyde and processed for light and electron microscopy. Our results showed that blood capillaries are initially in close proximity to the outer enamel epithelium. Between and intercalated with the capillaries are round/ovoid clusters of cells, some of which are vacuolated, closely apposed to the outer enamel epithelium. The outer enamel epithelium is not a continuous layer, but exhibits gaps between the cells. This suggests that the capillaries penetrate the enamel organ through these gaps, since no invagination of the epithelium was observed. The presence of a cluster of cells containing vacuoles suggests that vasculogenesis is taking place. Images showing loss of the basal lamina, proliferation of endothelial cells, presence of filopodia and lateral sprouting suggests that angiogenesis is also occurring. Thus, neoformation of capillaries of the molar enamel organ of rat seems to occur simultaneously by mechanisms of vasculogenesis and angiogenesis.     

The highly modified membrane of cornified cells in stratified squamous epithelia: A comparison of heterogenous deposits in keratinized and nonkeratinized epithelia

Summary The chemical nature of the thickened plasma membrane of cornified cells in stratified squamous epithelium was investigated in comparison with that in noncornified epithelium. Localizations of transglutaminase, molecular weight 92000 daltons, and detection of epidermal cysteine proteinase inhibitor were effected with a monoclonal antibody and a monospecific rabbit anti-inhibitor immunoglobulin, respectively, directed to the antigens. N-(7-dimethylamino-4-methylcoumarinyl) maleimide was used to demonstrate S-S cross-linking. In all keratinizing epithelia, the enzyme and inhibitor were deposited on membranes of granular cells. S-S bonds were formed in cornification with the appearance of electron-dense material by the inner leaflet. Both enzyme and inhibitors occurred on the corneal epithelium, but S-S linkage and the thickened plasma membrane did not form even at the last stage of maturation. On the other hand, the internal vaginal epithelium in the proestrous stage without keratinization contained the enzyme, but neither inhibitor nor S-S linkage. Both antigens and S-S bonds were detected when keratinization proceeded during estrus. The staining patterns in the epithelium near the vaginal introitus were identical to those in the skin. Cuboidal and simple epithelia exhibited none of those constituents. The findings indicated that heterogenous components contribute to modification of the plasma membrane of cornified cells, but S-S cross-linkages are associated exclusively with formation of the ultrastructurally unique membrane structure. In addition, findings suggested hormonal regulation in the chemical modification of the membrane in estrogen-sensitive internal vaginal epithelium.     

The postbranchial digestive tract of the ascidian, Polyandrocarpa misakiensis (Tunicata: Ascidiacea). 1. Oesophagus

The organization of the oesophagus in the budding styelid ascidian, Polyandrocarpa misakiensis, is described. The oesophagus consists of external and internal epithelium, and there are loose connective tissue, blood sinuses, and a muscular layer between them. The internal epithelium is simple columnar, except for the bottom of three folds. The external epithelium is simple squamous. The internal epithelium contains four cell types, i.e., ciliated mucous cells, band cells, endocrine cells, and undifferentiated cells. The ciliated mucous cells have apical cilia and microvilli, and two types of mucous vesicle. The band cells also have apical cilia and electron-dense granules in the apical cytoplasm. The endocrine cells are bottle-shaped, and have electron-dense granules both above and below the nucleus. The undifferentiated cells form pseudostratified epithelium at the bottom of each fold, and they have nuclei with prominent nucleoli. One type of coelomic cell, which has retractile cytoplasm, often migrates in the internal epithelium. Near the stomach, there are many darkly stained round cells clustered around the posterior end of the oesophagus. These two types of coelomic cells may be involved in the defense mechanism against the invasion of foreign organisms. The basic organization of the oesophagus of P. misakiensis is similar to those of other ascidians. However, the presence of three folds is a characteristic of a solitary species, rather than of a colonial species. Although ascidians are chordate invertebrates, the organization of their oesophagus is not very complex, which might reflect their life style.     

The seminal vesicle of the African catfish,Clarias gariepinus

Summary The seminal vesicle of the African catfish, Clarias gariepinus, consists of 36–44 fingerlike lobes built up of tubules in which a fluid is secreted containing acid polysaccharides, acid-, neutral- and basic proteins, and phospholipids. In this fluid sperm cells are stored. The seminal vesicle fluid immobilizes the sperm cells. After ejaculation, it prolongs the period of sperm activity. The seminal vesicle fluid is secreted by the epithelium lining the tubules. The tubules in the proximal part of the lobes are predominantly lined by a simple cylindrical and those of the distal part by a simple squamous epithelium. These epithelial cells contain enzymes involved in energy-liberating processes, the enzyme activites being proportional to the height of the cells. Interstitial cells between the tubules have enzyme-histochemical and ultrastructural features indicative of steroid biosynthesis. Similar characteristics are found in testicular interstitial cells. The most rostral seminal vesicle lobes and the most caudal testicular efferent tubules form a network of tubules that opens at the point where the paired parts of the sperm ducts fuse with each other. The tubules of most seminal vesicle lobes, however, form a complex system that fuses with the unpaired part of the sperm duct.     

The gingival epithelium of rodent molars with limited eruption

We have re-examined the arrangement of epithelia surrounding molar teeth with limited eruption in the rat, mouse and hamster. Our methods permitted enamel to be retained in paraffin and glycolmethacrylate sections, so providing optimal preservation of epithelial relationships. Three basic patterns of epithelial arrangement were observed on the various aspects of molars. Non-keratinized junctional epithelium covers the interdental septum and the base of all gingival crevices even in areas of gingival recession. In all areas other than the interdental septum, a fold of keratinized gingival epithelium lies beneath the lateral aspects of junctional epithelium. The buccal and lingual aspects of interdental septa show a zone of transition between these two basic types of epithelial arrangement. In intact gingiva the junctional epithelium can be seen to extend a considerable distance on to enamel surfaces, with the result that actual gingival crevice depths are even less than previously assumed. Artefacts resulting from loss of enamel during processing are discussed in the light of previous attempts to explain epithelial arrangements in rodent gingivae.     

Intermediate epithelium lining the mouse auditory tube

In the newborn mouse, the auditory tube is lined throughout the pharyngeal orifice to the tympanic orifice with ciliated columnar epithelium. In the adult mouse, the tube is divided into membranous and cartilaginous parts. The membranous part is covered by the ciliated columnar epithelium, while the cartilaginous part by varying types of epithelium ranging from ciliated columnar to stratified squamous type. It is suggested that the varying types of epithelium correspond to the 'intermediate epithelium', and that ciliated columnar epithelium transforms in part to stratified squamous epithelium by passing through the intermediate epithelium.