Ultrastructure of a cephalic sensory organ in larvae of the gastropod Phestilla sibogae (Aeolidacea, Nudibrachia)

The cephalic sensory organ is a superficial sensory receptor located between the velar lobes at the level of the shell aperture. Three cell types make up this sensory area: (1) six flask-shaped cells bearing numerous cilia; (2) adjacent supporting or accessory cells which have numerous, often branched, microvilli; and (3) vacuolated cells which occupy the center of the area. The flask-shaped cells appear to be the sensory units. These cells have a deep invaginated lumen, with cilia arising from the cell surface in the lumen oriented either toward the base of the lumen or toward the epidermal surface. These cilia, some of which extend slightly above the body surface, are presumed to be non-motile, as they lack (dynein?) arms on the axonemal A tubules and lack striated rootlets. The six flask cells are in intimate contact with the underlying cerebral ganglia and axons from each cell pass into ganglionic tissue. The supporting cells may be sensory, but no direct connection with the nervous system was seen. The function of the central vacuolated cells is not known. This cephalic organ may be a derivative of the original apical tuft of the trochophore stage.     

Formation and maturation of olfactory cilia monitored by odorant receptor-specific antibodies

The responsiveness of olfactory sensory neurons (OSNs) is based on odorant receptors (ORs) residing in the membrane of chemosensory cilia. It is still elusive as to when and how olfactory cilia are equipped with OR proteins rendering them responsive to odorants. To monitor the appearance of OR proteins in sensory compartments of OSNs, the olfactory epithelium of mice at various stages of prenatal development (lasting 19 days from conception) was investigated using immunohistochemical approaches and antibodies specific for different OR subtypes. These experiments uncovered that OR proteins accumulated in dendritic knobs of OSNs before the initiation of ciliogenesis (embryonic stage E12). As the first cilia were formed (E13), immunostaining in the knobs diminished. Cilia extended uprightly into the nasal cavity and were immunoreactive along the entire length, and particularly intense labeling was observed in expanded tips of cilia. During this phase of development (up to E18), the number of cilia per knob continuously increased. In the course of perinatal stages, longer cilia began to bend off and lie flat on the epithelial surface. The multiple cilia of a knob extended in length, and eventually the ciliary meshwork reached the characteristic complex pattern. In all stages, OR immunostaining was visible along the entire cilium. Thus, OR-specific antibodies allowed, for the first time, monitoring at the level of light microscopy the generation, outgrowth, and maturation of cilia in OSNs.     

Developmental expression of the calcium‐activated chloride channels TMEM16A and TMEM16B in the mouse olfactory epithelium

Calcium‐activated chloride channels are involved in several physiological processes including olfactory perception. TMEM16A and TMEM16B, members of the transmembrane protein 16 family (TMEM16), are responsible for calcium‐activated chloride currents in several cells. Both are present in the olfactory epithelium of adult mice, but little is known about their expression during embryonic development. Using immunohistochemistry we studied their expression in the mouse olfactory epithelium at various stages of prenatal development from embryonic day (E) 12.5 to E18.5 as well as in postnatal mice. At E12.5, TMEM16A immunoreactivity was present at the apical surface of the entire olfactory epithelium, but from E16.5 became restricted to a region near the transition zone with the respiratory epithelium, where localized at the apical part of supporting cells and in their microvilli. In contrast, TMEM16B immunoreactivity was present at E14.5 at the apical surface of the entire olfactory epithelium, increased in subsequent days, and localized to the cilia of mature olfactory sensory neurons. These data suggest different functional roles for TMEM16A and TMEM16B in the developing as well as in the postnatal olfactory epithelium. The presence of TMEM16A at the apical part and in microvilli of supporting cells is consistent with a role in the regulation of the chloride ionic composition of the mucus covering the apical surface of the olfactory epithelium, whereas the localization of TMEM16B to the cilia of mature olfactory sensory neurons is consistent with a role in olfactory signal transduction. © 2013 Wiley Periodicals, Inc. Develop Neurobiol 74: 657–675, 2014     

Presence of a Ciliary Patch in Preoral Epithelium of Sea Urchin Plutei

Removal of the hyaline layer from sea urchin embryos at the pluteus stage discloses a densely ciliated region in the preoral area of the ectodermal epithelium. In four-armed plutei, this ciliary path is located between the anterolateral arms and in eight-armed plutei it becomes surrounded by preoral and anterolateral arms. The area of the patch and the number of cilia increase with age. This patch is covered by cilia of unusual morphology and orientation. There are more than two cilia per cell which are coiled together several times around a small cone at the apical end of the cell. These coiled cilia run parallel to the surface of the cell but do not extend beyond the hyaline layer. The ciliary axoneme consists of a "9+2" microtubular structure, but no outer or inner dynein arms are observed. Although the cells with coiled cilia are present in a cluster constituting a part of the epithelium, they have axons that project from their basal (inner) ends. The structural characteistics of the ciliary patch suggest that it possesses a sensory function.     

Temporal and spatial distribution of ciliogenesis in the tracheobronchial airways of mice

Little is known about ciliogenesis as it proceeds through the entire airway tree, from the trachea to the terminal bronchioles, especially during the postnatal period. The purpose of this study was to define the spatial and temporal (prenatal and postnatal) pattern of normal cilia development in the mouse. Three airway generations representing the entire airway tree were examined: trachea, lobar bronchi, and terminal bronchiole. Ciliated cells in lung lobe whole mounts were labeled with a fluorescent dye for confocal microscopy, and ciliated cell surface density was measured for each airway generation and age. The same samples were examined by scanning electron microscopy to verify the appearance of ciliated cells among the differentiating epithelium of the airways. Ciliated cells were first detected in the trachea and lobar bronchi at 16 days gestational age (DGA) and in the terminal bronchioles at 18 DGA. Ciliated cell surface density increased with prenatal and postnatal age at all airway levels. However, the ciliated cell surface density of the trachea and lobar bronchi was always greater compared with the terminal bronchiole. In conclusion, the study revealed that in developing tracheobronchial airways of the mouse: 1) Ciliogenesis differs temporally and spatially by airway generation; 2) Ciliated cell surface density increases with age in all airway generations, but density decreases in a proximal to distal direction; and 3) A significant portion of ciliogenesis continues after birth. This study provides a healthy basis for investigations of neonatal pulmonary disease or pollutant toxicity affecting cilia and its functions.     

A novel population of neuronal cells expressing the olfactory marker protein (OMP) in the anterior/dorsal region of the nasal cavity

The olfactory marker protein (OMP) is expressed in mature chemosensory neurons in the nasal neuroepithelium. Here, we report the identification of a novel population of OMP-expressing neurons located bilaterally in the anterior/dorsal region of each nasal cavity at the septum. These cells are clearly separated from the regio olfactoria, harboring the olfactory sensory neurons. During mouse development, the arrangement of the anterior OMP-cells undergoes considerable change. They appear at about stage E13 and are localized in the nasal epithelium during early stages; by epithelial budding, ganglion-shaped clusters are formed in the mesenchyme during the perinatal phase, and a filiform layer directly underneath the nasal epithelium is established in adults. The anterior OMP-cells extend long axonal processes which form bundles and project towards the brain. The data suggest that the newly discovered group of OMP-cells in the anterior region of the nasal cavity may serve a distinct sensory function.     

Appearance of B and H blood group antigens in the developing cochlear hair cells

Summary The presence of human blood-group antigens was analyzed in the rat cochlea during its postnatal development, using anti-A, anti-B and anti-H antibodies. At no stage was reactivity with anti-A antibody observed. With the anti-H antibody, a strong reactivity was observed from 1 to 9 days after birth within hair cells and some other surface epithelial cells of the cochlear duct. After postnatal day 9, only a faint reactivity persisted in a few non-sensory cells. With the anti-B antibody, only hair cells were selectively labeled. At early stages (postnatal day 1 and 3), the reactivity was intense and observed both around the cell surface and within the supranuclear region of cytoplasm. Later on, the reactivity decreased; it was limited at postnatal day 9 to a reactive spot below the cuticular plate. Results are compared with a preliminary finding describing the first appearance of B and H antigens in the organ of Corti at a prenatal stage, and with data concerning other sensory and neural structures. The appearance and progressive disappearance of B and H antigens on sensory and non-sensory cells can be correlated with significant events in the development of the cochlea. The transient expression of B and H antigens in cochlear sensory cells may correspond to developmental changes in their surface glycoconjugates.     

Surface specializations of the olfactory epithelium of rainbow trout, Salmo gairdneri

Receptors for olfactory stimulus molecules appear to be located at the surface of olfactory receptor cells. The ultrastructure of the distal region of rainbow trout (Salmo gairdneri) olfactory epithelium was examined by transmission electron microscopy. On the sensory olfactory epithelium, which occurs in the depressions of secondary folds of the lamellae of the rosettes, five cell types were present. Type I cells have a knob-like apical projection which is unique in this species because it frequently contains cilia axonemes within its cytoplasm in addition to being surrounded by cilia. Type II cells bear many cilia oriented unidirectionally on a wide, flat surface. Type III cells have microvilli on a constricted apical surface and centrioles in the subapical cytoplasm. Type IV cells contain a rod-like apical projection filled with a bundle of filaments, and type V cells are supporting cells. Cilia on the sensory epithelium contain the 9 + 2 microtubule fiber pattern. Dynein arms are clearly present on the outer doublet fibers, which suggests that the cilia in the olfactory region are motile. Their presence in olfactory cilia of vertebrates has been controversial. The cilia membrane in this species is unusual in often showing outfoldings, within which are included small, irregular vesicles or channels. In addition, cilia on type II cells frequently contain dense-staining bodies closely apposed to the membranes, along with a densely stained crown at the cilia tip. Previous biochemical evidence indicates that odorant receptors are associated with the cilia.     

Ultrastructural investigation of the nuchal organs ofPygospio elegans (Polychaeta). I. Larval nuchal organs

The structural differentiation of the nuchal organs during the post-embryonic development ofPygospio elegans is described. The sensory organs are composed of two cell types: ciliated cells and bipolar primary sensory cells, constituting the nuchal ganglion, which is associated with both the sensory epithelium and the brain. Since the sensory neurons are largely integrated into posterolateral parts of the cerebral ganglion, the nuchal organs are primary presegmental structures. The microvilli of the ciliated cells form a cover over the cuticle with a presumed protective function. An extracellular space extends between cuticle and sensory epithelium. The distal dendrites of the sensory cells terminate in sensory bulbs, bearing one modified sensory cilium each that projects into the olfactory chamber, embedded within the secretion of the ciliated cells. During development, the nuchal organs increase in size. This is accompanied by a shift in position, an expansion of the sensory area, and secretory activity of the ciliated cells. The nuchal ganglion differentiates into three nuchal centres forming three distinct sensory areas around the ciliated region. Each nuchal complex reveals two short nuchal nerves comprising the sensory axons, which enter the posterior circumesophageal connective. The sensory cells lying in the brain exhibit neurosecretory activity; the sensory cilia enlarge their surface area by dilating and branching. Nuchal organs accomplish the basic structural adaptions of chemoreceptors and show structural analogies to arthropod olfactory sensilla; thus, there is every reason to suppose chemoreceptor function.     

Scanning electron microscopy of the channel catfish olfactory lamellae

The olfactory lamellae of the channel catfish (Ictalurus punctatus) are composed of sensory and indifferent (non-sensory) epithelia organized into two distinct regions on both surfaces of each lamella. The smaller sensory region located adjacent to the midline raphe has fewer cilia per unit surface area than the indifferent epithelium and contains the olfactory neurons whose ciliated dendritic terminals occur at the epithelial surface. The indifferent epithelium, comprising the greater surface area of the olfactory lamella, is covered with a dense mat of non-sensory cilia. Fractured carbon dioxide critical point dried lamellar tissue revealed the underlying cellular structure. The lamellae are composed of two layers of epithelium enclosing a thin stromal layer. Olfactory receptors were observed in the fractured tissue only within the sensory epithelium.     

Meprin mRNA in rat intestine during normal and glucocorticoid-induced maturation: divergent patterns of expression of alpha and beta subunits

Meprin is a zinc-metalloendopeptidase expressed in intestinal epithelial cells. In rat jejunum collected from postnatal day 4 (P4) through P25 meprins alpha mRNA exhibited uniform levels for the first three postnatal weeks and then declined, whereas meprin beta mRNA showed a biphasic pattern with high levels in the first postnatal week followed by low levels from P7 through P19 and then a marked rise at P22 and P25. Dexamethasone treatment beginning at P10 had no significant effect on levels of meprins a mRNA, whereas this treatment caused a precocious increase in expression of meprin beta mRNA. These divergent patterns of expression of meprins alpha and beta mRNA suggest distinct roles for the two subunits during the suckling and weaning phases of rodent intestinal development.     

Putative sensory structures in marine bryozoans

Abstract. SEM studies of 21 species of marine bryozoans demonstrated that the abfrontal side of the tentacles bears a row of mono- or multiciliated cells, which are presumably sensory. In stenolaemates, the abfrontal cells, as well as the cells at the tentacle tips and the laterofrontal cells, are monociliated. In the 17 gymnolaemate species studied, each tentacle tip bears at least 3 multiciliated cells, each with a tuft of 5–7 stiff cilia of various lengths. On the abfrontal tentacle surface, mono- and multiciliated cells alternate, but all species studied have multiciliated cells at the base and the tip of each tentacle. In live animals, single cilia perform occasional flicks, whereas the tufts of 7–15 cilia on the multiciliated cells are immotile. Length and number of abfrontal cilia vary between species. Two types of multiciliated, putative sensory organs were found on the introvert of some gymnolaemates. One has an apical knob surrounded by a ring of cilia; the other has an apical tuft of cilia. The ultrastructure of the sensory cells of tentacles and introvert was studied in Rhamphostomella ovata . Our observations on both fixed and living material all suggest that these cells are primitive mechanoreceptors. The few species lacking ciliary structures on the introvert have long proximal ciliary tufts on the abfrontal tentacle surface.     

Fine structure of the cephalic sensory organ in veliger larvae of Littorina littorea, (L.) (Mesogastropoda,Littorinidae)

The cephalic sensory organ (CSO) in planktonic veliger larvae of Littorina littorea is situated dorsally between the velar lobes at the level of the shell aperture. It consists of ciliated primary sensory cells, adjacent accessory cells and supporting epithelial cells. Cell bodies of the ciliated cells originate in the cerebral commissure and their dendrites pass to the epidermis. The flask-shaped sensory cells are characterized by a deep invaginated lumen with modified cilia arising from the cell surface in the lumen. These cilia are presumed to be non-motile because they lack striated rootlets and show a modified microtubular pattern (6 + 2, 7 + 2 and 8 + 2). The adjacent accessory cells never possess an invaginated lumen; occasionally cilia and branched microvilli arise from the apical surface. These cells may be sensory, but there is no obvious direct connection with the nervous system. The supporting epithelial cells are part of the epidermis and flank the apical necks of the sensory and accessory cells. Morphological evidence suggests that the CSO may function in chemoreception related to substrate selection at settlement, feeding or other behaviour.     

Deletion of IFT20 in the mouse kidney causes misorientation of the mitotic spindle and cystic kidney disease

Primary cilia project from the surface of most vertebrate cells and are thought to be sensory organelles. Defects in primary cilia lead to cystic kidney disease, although the ciliary mechanisms that promote and maintain normal renal function remain incompletely understood. In this work, we generated a floxed allele of the ciliary assembly gene Ift20. Deleting this gene specifically in kidney collecting duct cells prevents cilia formation and promotes rapid postnatal cystic expansion of the kidney. Dividing collecting duct cells in early stages of cyst formation fail to properly orient their mitotic spindles along the tubule, whereas nondividing cells improperly position their centrosomes. At later stages, cells lacking cilia have increased canonical Wnt signaling and increased rates of proliferation. Thus, IFT20 functions to couple extracellular events to cell proliferation and differentiation.     

Functional nasal morphology of chimaerid fishes

Holocephalans (chimaeras) are a group of marine fishes comprising three families: the Callorhinchidae (callorhinchid fishes), the Rhinochimaeridae (rhinochimaerid fishes) and the Chimaeridae (chimaerid fishes). We have used X‐ray microcomputed tomography and magnetic resonance imaging to characterise in detail the nasal anatomy of three species of chimaerid fishes: Chimaera monstrosa, C. phantasma and Hydrolagus colliei. We have shown that the nasal chamber of these three species is linked to the external environment by an incurrent channel and to the oral cavity by an excurrent channel via an oral groove. A protrusion of variable morphology is present on the medial wall of the incurrent channel in all three species, but is absent in members of the two other holocephalan families that we inspected. A third nasal channel, the lateral channel, functionally connects the incurrent nostril to the oral cavity, by‐passing the nasal chamber. From anatomical reconstructions, we have proposed a model for the circulation of water, and therefore the transport of odorant, in the chimaerid nasal region. In this model, water could flow through the nasal region via the nasal chamber or the lateral channel. In either case, the direction of flow could be reversed. Circulation through the entire nasal region is likely to be driven primarily by the respiratory pump. We have identified several anatomical features that may segregate, distribute, facilitate and regulate flow in the nasal region and have considered the consequences of flow reversal. The non‐sensory cilia lining the olfactory sensory channels appear to be mucus‐propelling, suggesting that these cilia have a common protective role in cartilaginous fishes (sharks, rays and chimaeras). The nasal region of chimaerid fishes shows at least two adaptations to a benthic lifestyle, and suggests good olfactory sensitivity, with secondary folding enhancing the hypothetical flat sensory surface area by up to 70%. J. Morphol. 274:987–1009, 2013. © 2013 Wiley Periodicals, Inc.     

Development of the hypothalamic luteinizing hormone-releasing hormone-containing neuron system in the rat: in vivo and in transplantation studies

The development of the hypothalamic LHRH-containing neuron system was immunohistochemically investigated in vivo and in tissue transplantation using rat embryos aged from 12.5 to 17.5 days of gestation. The sera used were generated against rat gonadotropic hormone-releasing hormone-associated peptide (28-56) (rGAP) and LHRH. Immunoreaction for rGAP was first found in cells migrated from and in the vomeronasal organ on Days 13.5 and 14.5 of gestation. Immunoreactive cells seem to ascend along the terminal nerves, reaching the medial surface of the forebrain vesicles. Subsequently the cells occurred in the septum and further into their final position in the septopreoptic-diagonal band area on Days 16.5-17.5 of gestation; during this traverse the cells become secretory neurons after changes in morphology and in behavior. Intraventricular transplantation revealed that nasal epithelia of Day 12.5 embryos raised only a few cells immunoreactive both for LHRH and rGAP, but a great number of immunoreactive cells and fibers in the presence of the medial basal hypothalamus (MBH). The fibers formed a median eminence-like structure together with dense capillary plexus that had grown in the cografted MBH. The same phenomenon was apparently observed in the grafts obtained from older embryos of gestation, but not in the combined grafts of the anterior septum and the nasal epithelium or the MBH. We conclude that hypothalamic LHRH neurons originate from the nasal placode and acquire secretory behavior in the presence of the MBH.     

Proteomic analysis of a membrane preparation from rat olfactory sensory cilia

The cilia of mammalian olfactory receptor neurons (ORNs) represent the sensory interface that is exposed to the air within the nasal cavity. The cilia are the site where odorants bind to specific receptors and initiate olfactory transduction that leads to excitation of the neuron. This process involves a multitude of ciliary proteins that mediate chemoelectrical transduction, amplification, and adaptation of the primary sensory signal. Many of these proteins were initially identified by their enzymatic activities using a membrane protein preparation from olfactory cilia. This so-called "calcium-shock" preparation is a versatile tool for the exploration of protein expression, enzyme kinetics, regulatory mechanisms, and ciliary development. To support such studies, we present a first proteomic analysis of this membrane preparation. We subjected the cilia preparation to liquid chromatography-electrospray ionisation (LC-ESI-MS/MS) tandem mass spectrometry and identified 268 proteins, of which 49% are membrane proteins. A detailed analysis of their cellular and subcellular localization showed that the cilia preparation obtained by calcium shock not only is highly enriched in ORN proteins but also contains a significant amount of nonciliary material. Although our proteomic study does not identify the entire set of ciliary and nonciliary proteins, it provides the first estimate of the purity of the calcium-shock preparation and provides valuable biochemical information for further research.     

Candidate chemosensory cells in the stomach mucosa of young postnatal mice during the phases of dietary changes

The complex physiology of the gastrointestinal (GI) tract must permanently be adjusted according to the composition of ingested food, which requires continuous monitoring by appropriate sensory systems. Sensing the dietary constituents is thought to be mediated by chemosensory cells residing in the mucosa of the GI tract. We have examined the appearance and differentiation of candidate chemosensory cells at distinct postnatal stages and visualized cells that express gustducin or TRPM5. Two critical stages have been considered: the suckling period when the neonates are nourished exclusively on milk and the weaning period when the diet gradually changes to solid food. At early postnatal stages, only a few gustducin- or TRPM5-expressing cells have been found; they display an immature morphology. At the time of weaning, numerous gustducin- or TRPM5-positive cells are present in the gastric mucosa and are isomorphic to adult candidate chemosensory cells. The typical accumulation of gustducin and TRPM5 cells at the border between the forestomach and corpus region and the characteristic tissue fold or “limiting ridge” have not been observed at early postnatal stages but are complete at the time of weaning. The appearance of candidate chemosensory cells at the strategic position occurs within the last few days before weaning but after the formation of the limiting ridge. Thus, both the topographic arrangement of the cells and the limiting ridge seem to be important features for the processing of solid food in the mouse stomach.     

Role of gastrin in the development of gastric mucosa,ECL cells and A-like cells in newborn and young rats

Histamine-producing ECL cells and ghrelin-producing A-like cells are endocrine/paracrine cell populations in the acid-producing part of the rat stomach. While the A-like cells operate independently of gastrin, the ECL cells respond to gastrin with mobilization of histamine and chromogranin A (CGA)-derived peptides, such as pancreastatin. Gastrin is often assumed to be the driving force behind the postnatal development of the gastric mucosa in general and the ECL cells in particular. We tested this assumption by examining the oxyntic mucosa (with ECL cells and A-like cells) in developing rats under the influence of YF476, a cholecystokinin-2 (CCK(2)) receptor antagonist. The drug was administered by weekly subcutaneous injections starting at birth. The body weight gain was not affected. Weaning occurred at days 15-22 in both YF476-treated and age-matched control rats. Circulating gastrin was low at birth and reached adult levels 2 weeks after birth. During and after weaning (but not before), YF476 greatly raised the serum gastrin concentration (because of abolished acid feedback inhibition of gastrin release). The weight of the stomach was unaffected by YF476 during the first 2-3 weeks after birth. From 4 to 5 weeks of age, the weight and thickness of the gastric mucosa were lower in YF476-treated rats than in controls. Pancreastatin-immunoreactive cells (i.e. all endocrine cells in the stomach) and ghrelin-immunoreactive cells (A-like cells) were few at birth and increased gradually in number until 6-8 weeks of age (control rats). At first, YF476 did not affect the development of the pancreastatin-immunoreactive cells, but a few weeks after weaning, the cells were fewer in the YF476 rats. The ECL-cell parameters (oxyntic mucosal histamine and pancreastatin concentrations, the histidine decarboxylase (HDC) activity, the HDC mRNA levels and serum pancreastatin concentration) increased slowly until weaning in both YF476-treated and control rats. From then on, there was a further increase in the ECL-cell parameters in control rats but not in YF476 rats. The postnatal development of the ghrelin cells (i.e. the A-like cells) and of the A-like cell parameters (the oxyntic mucosal ghrelin concentration and the serum ghrelin concentrations) was not affected by YF476 at any point.We conclude that gastrin affects neither the oxyntic mucosa nor the endocrine cells before weaning. After weaning, CCK(2) receptor blockade is associated with a somewhat impaired development of the oxyntic mucosa and the ECL cells. While gastrin stimulation is of crucial importance for the onset of acid secretion during weaning and for the activation of ECL-cell histamine formation and secretion, the mucosal and ECL-cell growth at this stage is only partly gastrin-dependent. In contrast, the development of the A-like cells is independent of gastrin at all stages.     

Superficial cell differentiation during embryonic and postnatal development of mouse urothelium

After drastic urothelial destruction around birth and around postnatal day 6, mouse urothelial renewal starts each time de novo. The differentiation of superficial cells during urothelial restoration was followed for the first time from embryonic day 15 to postnatal day 6 by the detection of differentiation markers: cytokeratins, uroplakins and apical membrane specialization. The differentiation markers of short-lived superficial cells were studied before and after urothelial destruction. Three distinctive types of superficial cells, typical for certain developmental period, were characterised: cells at low differentiation stage with microvilli and cilia, expressing CK7 and CK18, detected on embryonic day 15; cells at advanced differentiation stage with star-like arrangement of prominent membrane ridges, expressing CK7 and CK20, present between the two urothelial destruction events; highly differentiated cells with typically jagged apical surface, expressing CK7 and CK20, found twice during development. This cell type appears for the first time on embryonic day 18 as the terminal stage of embryonic differentiation. It was found again on postnatal day 6 as an initial stage of differentiation, leading toward terminally differentiated cells of the adult urothelium. Our work proves that apical membrane specialization is the most valuable differentiation marker of superficial cells.