Light microscopy and scanning electron microscopy studies on the reduction of the tongue microstructures in the white stork (Ciconia ciconia,Aves)

The structure of the tongue in the white stork (Ciconia ciconia) is observed macroscopically and under light and scanning electron microscopy. Our observations of the tongue reveal a rare terminal reduction of the size of the tongue and microstructures of the lingual mucosa among the investigations of birds published so far. The short, triangular tongue with a pointed tip is approximately 2.5 cm long in the adult and is situated in the caudal part of the oral cavity close to the laryngeal prominence. On the dorsal surface of the tongue, no typical mucosa microstructures like lingual papillae, median groove or lingual prominence are observed. The main structure of the tongue is composed of rostral part of hyoid apparatus, that is, entoglossal cartilage connects with basihyoid. Very thin mucosa is composed of fibrous connective tissue covered with orthokeratinized epithelium. No lingual glands and muscles are observed in the lamina propria of mucosa. Even though the triangular shape of the tongue in the white stork is typical for birds, the inner structure of the reduced organ is composed only of flat cartilagineous entoglossum of hyoid apparatus. During feeding behaviour of the white stork, the food transportation in oral cavity called cranio‐inertial transport is undoubtedly affected by structural reduction of the tongue.     

Confocal evaluation of native and induced lectin binding contributes to discriminate between lingual gland glycocomponents in quail

A confocal analysis was performed on the quail (Coturnix coturnix japonica) lingual salivary glands where the carbohydrate chains were studied by lectin histochemistry. For this purpose, appropriate FITC- and TRITC-conjugates were used for double binding also accomplished with sialidase digestion. The glycosidic components of the quail lingual salivary glands were found to be heterogeneously distributed on the different secretory structures as well as on the single secretory elements of each adenomere. The rostral portion of the anterior lingual gland was found to only secrete neutral glycocomponents, characterized by terminal beta-galactose, N-acetylgalactosamine and fucose residues in contrast to the caudal portion that was shown to be extremely heterogeneous and to produce sialylated glycoconjugates characterized by the terminal sequences sialic acid-beta-galactose-N-acetylgalactosamine, sialic acid-beta-galactose-N-acetylglucosamine, and sialic acid-alpha-N-acetylgalactosamine partly codistributed within secretory adenomeres. The posterior lingual gland was observed to be the major contributor to the secretion of salivary mucins containing sialoglycoconjugates with terminal sialic acid residues linked to beta-galactose-N-acetylgalactosamine or alpha-N-acetylgalactosamine often located in distinct secretory elements.     

Connexin 43 Is Necessary for Salivary Gland Branching Morphogenesis and FGF10-induced ERK1/2 Phosphorylation

Cell-cell interaction via the gap junction regulates cell growth and differentiation, leading to formation of organs of appropriate size and quality. To determine the role of connexin43 in salivary gland development, we analyzed its expression in developing submandibular glands (SMGs). Connexin43 (Cx43) was found to be expressed in salivary gland epithelium. In ex vivo organ cultures of SMGs, addition of the gap junctional inhibitors 18α-glycyrrhetinic acid (18α-GA) and oleamide inhibited SMG branching morphogenesis, suggesting that gap junctional communication contributes to salivary gland development. In Cx43^−/− salivary glands, submandibular and sublingual gland size was reduced as compared with those from heterozygotes. The expression of Pdgfa, Pdgfb, Fgf7, and Fgf10, which induced branching of SMGs in Cx43^−/− samples, were not changed as compared with those from heterozygotes. Furthermore, the blocking peptide for the hemichannel and gap junction channel showed inhibition of terminal bud branching. FGF10 induced branching morphogenesis, while it did not rescue the Cx43^−/− phenotype, thus Cx43 may regulate FGF10 signaling during salivary gland development. FGF10 is expressed in salivary gland mesenchyme and regulates epithelial proliferation, and was shown to induce ERK1/2 phosphorylation in salivary epithelial cells, while ERK1/2 phosphorylation in HSY cells was dramatically inhibited by 18α-GA, a Cx43 peptide or siRNA. On the other hand, PDGF-AA and PDGF-BB separately induced ERK1/2 phosphorylation in primary cultured salivary mesenchymal cells regardless of the presence of 18α-GA. Together, our results suggest that Cx43 regulates FGF10-induced ERK1/2 phosphorylation in salivary epithelium but not in mesenchyme during the process of SMG branching morphogenesis.     

EGF-receptor regulates salivary gland branching morphogenesis by supporting proliferation and maturation of epithelial cells and survival of mesenchymal cells

Epidermal growth factor receptor (EGF-R) regulates epithelial morphogenesis during development and is important for the proper branching of the lung, mammary gland, and pancreas. We analyzed the salivary gland phenotype of EGF-R-deficient mice and showed impaired growth, branching, and maturation of the epithelium. Furthermore, treatment of wild-type E13 salivary glands with gefitinib, a small molecular inhibitor of EGF-R, led to apoptosis of the mesenchyme. Interestingly, MMP2 and plasminogen activators were upregulated upon inhibition of EGF-R signaling. To summarize, we show that EGF-R is a physiological regulator of salivary gland development and its main function is to support the proliferation and maturation of the epithelium and the survival of the mesenchyme.     

Salivary gland development

Salivary glands provide an excellent model for the study of epithelial-mesenchymal interactions and branching morphogenesis. This review will discuss the anatomy of different types of glands, in a range of different organisms. Then, concentrating on the mouse submandibular gland, the stages of salivary gland development will be reviewed and the relative role of the mesenchyme and the epithelium will be discussed. Finally, the genes thought to play a role in development of the glands from initiation to differentiation will be reviewed.     

The histological structure and histochemistry of the mucosa of the nasal conchae in geese,Anser anser

We investigated the histological structure and histochemistry of the nasal conchae of geese and compared these structures with those of other avian species. The rostral, middle and caudal conchae were dissected from the nasal cavity of eight geese, fixed in Carnoy’s solution and embedded in paraffin. The entrance of the rostral concha was lined by keratinized stratified squamous epithelium, which toward the middle concha was replaced by modified keratinized squamous epithelium, the deep layer of which opened into tubular glandular structures containing secretory epithelium on crypt-like invaginations. The lamina propria of the rostral concha contained numerous Grandry’s and Herbst corpuscles, which are pressure-sensitive receptors peculiar to waterfowl. The lamina propria of the middle concha contained solitary lymphoid follicles and lymphocyte infiltrations. The cartilaginous component of the middle concha was highly convoluted and resembled a spiral of two and a half scrolls, which were lined by pseudostratified columnar epithelium. We observed that unlike mammals, this epithelium contained mostly intraepithelial alveolar glands rather than goblet cells. The caudal concha was similar to the middle concha, but less convoluted. It was lined by olfactory epithelium and its lamina propria contained serous Bowman’s glands as well as olfactory nerve fibers. Histochemical examination demonstrated that while none of the conchae contained sulfated mucins, except for the cartilage, the intraepithelial glands of the rostral and middle conchae contained mostly carboxylated acidic mucin and some neutral mucin, and were thus of the mixed type. The outermost scroll of the spiral of the middle concha contained some periodate-Schiff stained mucins. Of the glands of the mucosa of the middle concha, the deep tubuloalveolar glands in the convex parts of the scrolls contained primarily acidic mucins, while the shallow intraepithelial alveolar glands in the concave parts of the scrolls contained primarily neutral mucins. Our findings indicate that the rostral and caudal conchae primarily have a sensory function and the middle concha participates in mucosal defense.     

Human salivary gland branching morphogenesis: morphological localization of claudins and its parallel relation with developmental stages revealed by expression of cytoskeleton and secretion markers

Development of salivary glands is a highly complex and dynamic process termed branching morphogenesis, where branched structures differentiate into mature glands. Tight junctions (TJ) are thought to play critical roles in physiological functions of tubular organs, contributing to cell polarity and preventing lateral movement of membrane proteins. Evidence demonstrated that claudins are directly involved in TJ formation and function. Using immunohistochemistry and immunofluorescence we have mapped the distribution of claudins-1, 2, 3, 4, 5, 7 and 11 and compared it with the expression of differentiation markers in human salivary glands obtained from foetuses ranging from weeks 4 to 24 of gestation. Expression of all claudins, except claudin-2 was detected in the various phases of human salivary gland development, up to fully mature salivary gland. The expression of all claudins increased according to the progression of salivary gland maturation evidenced by the classical markers-cytokeratin 14, cytokeratin low molecular weight, smooth muscle actin and human secretory component. Tight junction proteins-claudins appear to be important in the final shape and physiological functions of human salivary glands and are parallel related with markers of salivary gland differentiation.     

An ultrastructural analysis of the salivary system of the terrestrial mollusc, Limax maximus.

The bilateral salivary glands, ducts, and nerves of the giant garden slug Limax maximus control the secretion of saliva and its transport to the buccal mass. Each salivary nerve, which originates at the buccal ganglion, contains over 3000 axon profiles. The axons innervate the musculature of the duct and branch within the gland. The salivary duct is composed of several muscular layers surrounding an epithelial layer which lines the duct lumen. The morphology of the duct epithelium indicates that it may function in ion or water balance. The salivary gland contains four major types of secretory cells. The secretory products are released from vacuoles in the gland cells, and are presumably transported by cilia in the collecting ducts of the gland into the larger muscular ducts.     

Autonomic innervation of the salivary glands in cebid monkeys: a histochemical study.

The autonomic innervation of the major and minor salivary glands was studied in five species of cebid monkeys using acetylcholinesterase (AChE) and catecholamine histochemistry. Catecholamine-containing and AChE-positive nerve fibres were observed in the vessels and secretory endpieces of all glands, with no apparent predominance of one type over the other. In the intralobular ducts, however, the cholinergic innervation predominates. In the major salivary and minor sublingual glands the density of the nervous supply was higher, whereas in the secondary mandibular and posterior lingual glands it was less dense. The morphological patterns of salivary gland innervation found in Cebidae are compared with those of the related family Callitrichidae.     

A high resolution sem study of human minor salivary glands

By SEM we have investigated the human minor salivary glands using the NaOH method for the visualization of endpieces and myoepithelial cells, and the osmium maceration technique that reveals membranous intracellular structures. With the former method all minor glands, including the posterior deep (Ebner's) lingual glands, consist of tubules sometimes dilated into alveoli, while true acini of the kind observed in human major salivary glands, are absent. Tubules of the posterior deep lingual gland exhibit stellate myoepitelial cells that leave a substantial part of the secretory cells uncovered. The latter cells, at variance with serous cells of major glands, do not show basal folds. In contrast, tubules of the other minor glands, like the mucous ones of major glands, are covered almost completely by band-like myoepithelial cells. The osmium maceration method clearly demonstrates that posterior deep lingual glands are serous in character and that all the other minor glands, together with the predominant mucous cells, possess a variable number of seromucous cells that, despite variations among individuals, increase in order from palatine and posterior superficial lingual (Weber's), to minor sublingual, labial, anterior lingual (Blandin and Nuhn's), and buccal glands.     

Morphological, micromorphological and histochemical studies on the parotid salivary glands of the one-humped camel (Camelus dromedarius).

The morphology, blood and nerve supply of the parotid salivary glands of the one-humped camel were studied in detail. The intraglandular portion of the duct system was also examined. The histological and histochemical studies showed that the parotid salivary glands of the camel are of the tubuloacinar type and are serumocoid in nature. The secretory acini and tubules show themselves in 3 different forms according to the different phases of their secretory cycle. The duct system of the gland contains goblet cells between its lining epithelium. The intercalated ducts show ampullation followed by narrowing that help in mixing the secretion. Intraepithelial glands are found in the terminal part of the parotid duct.     

THE DEVELOPMENT OF SURFACE SPECIALIZATION IN THE SECRETORY EPITHELIUM OF THE AVIAN SALT GLAND IN RESPONSE TO OSMOTIC STRESS

Cell surface specialization, a characteristic common to most ion-transporting epithelia, was studied in the salt (nasal) gland of the domestic duck in relation to osmotic stress. Three days after hatching, experimental ducklings were given 1% NaCl to drink for 12 hr and freshwater for the remainder of each day. Control ducklings were maintained exclusively on freshwater. The fine structure of the secretory epithelium was examined on various days of the regimen. The nasal gland epithelium of the secretory lobule is composed of several types of cells. Peripheral cells, lying at the blind ends of the branched secretory tubules, are similar in both control and experimental animals at all stages of glandular development. These generative cells contain few mitochondria and have nearly smooth cell surfaces. Partially specialized secretory cells predominate in the secretory tubules of control animals and appear as transitional cells in the tubular epithelium of salt-stressed animals. These cells contain few mitochondria and bear short folds along their lateral cell surfaces. Fully specialized cells dominate the secretory epithelium of osmotically stressed ducklings. The lateral and basal surfaces of these cells are deeply folded, forming complex intra- and extracellular compartments. This vast increase in absorptive surface area is paralleled by an increase in the number of mitochondria that pack the basal compartments. The development of this fully specialized cell is correlated with the marked increase in (Na⁺-K⁺)-ATPase activity in the glands of osmotically stressed birds.     

Ultrastructure of human labial salivary glands. 3. Myoepithelium and ducts

Myoepithelial cells were present between the basal lamina and the acinar secretory cells of human labial salivary glands. In form and disposition, they resembled myoepithelial cells in the major salivary glands. Many of these cells possessed single cilia on their upper surfaces. Such cilia occasionally extended into invaginations of the overlying secretory cell. The intercalated ducts were variable in occurrence. Their epithelium ranged from columnar to squamous, and showed few signs of secretory activity. Few intralobular ducts possessed basal striations. While mitochondria were abundant in non-striated cells, they were randomly disposed in both basal and apical cytoplasm, and the basal plasmalemma showed only occasional infoldings. The paucity of true striated ducts in labial salivary glands may be responsible for the high concentration of sodium and chloride in unstimulated labial gland salivary secretions.     

Structural simplicity of theZonula Occludens in the electrolyte secreting epithelium of the avian salt gland

Summary The structure of thezonula occludens in the secretory epithelium of the salt gland of the domestic duck was determined by thin section and freeze-fracture electron microscopy. These glands secrete an effluent with a NaCl concentration four times that of plasma, and thus maintain a steep ionic gradient across their secretory epithelium. Freezefracture replicas from salt stressed ducks demonstrate that thezonula occludens is surprisingly shallow in depth (20–25 nm) and generally consists of two parallel junctional strands which are juxaposed along their entire length. In addition to the simplicity of the junction separating mucosal and serosal compartments, the ratio of junctional length to apical surface area is large since luminal surfaces of secretory cells are narrow and intermesh with one another. Thezonula occludens in nonsecreting fresh water-adapted birds is similar to the salt stressed group except that two sets of double strand junctions are seen in addition to junctions consisting of a single set. Based on previous ultrastructural, cytochemical and physiological studies in salt glands and in other epithelia, a model for salt secretion was suggested in which intercellular space Na⁺, generated by basolateral ouabain-sensitive Na⁺ pumps, reaches the lumen via a paracellular route (Ernst & Mills, 1977,J. Cell Biol. 75:74). The simplicity of the morphological appearance of thezonula occludens in the salt gland, which resembles that described for several epithelia known to be leaky to ions, is consistent with this hypothesis.     

Immunolocalization patterns of cytokeratins during salivary acinar cell development in mice

Embryonic development of the mouse salivary glands begins with epithelial thickening and continues with sequential changes from the pre-bud to terminal bud stages. After birth, morphogenesis proceeds, and the glands develop into a highly branched epithelial structure that terminates with saliva-producing acinar cells at the adult stage. Acinar cells derived from the epithelium are differentiated into serous, mucous, and seromucous types. During differentiation, cytokeratins, intermediate filaments found in most epithelial cells, play vital roles. Although the localization patterns and developmental roles of cytokeratins in different epithelial organs, including the mammary glands, circumvallate papilla, and sweat glands, have been well studied, their stage-specific localization and morphogenetic roles during salivary gland development have yet to be elucidated. Therefore, the aim of this study was to determine the stage and acinar cell type-specific localization pattern of cytokeratins 4, 5, 7, 8, 13, 14, 18, and 19 in the major salivary glands (submandibular, sublingual, and parotid glands) of the mouse at the E15.5, PN0, PN10, and adult stages. In addition, cell physiology, including cell proliferation, was examined during development via immunostaining for Ki67 to understand the cellular mechanisms that govern acinar cell differentiation during salivary gland morphogenesis. The distinct localization patterns of cytokeratins in conjunction with cell physiology will reveal the roles of epithelial cells in salivary gland formation during the differentiation of serous, mucous or seromucous salivary glands.     

Configuration of the medial and lateral segments of duck (Anas platyrhynchos) salt glands

Salt glands of the domestic duck Anas platyrhynchos differ from those of the herring gull Larus argentatus and other birds. In ducks, each salt gland consists of distinct medial and lateral segments. Centrally located drainage ducts that extend along the entire length of these medial and lateral segments collect hypertonic fluid secreted by an array of lobules. Each lobule is formed by a single mass of branched tubules in which the direction of capillary blood flow is opposite to that of the secreted fluid. This fluid drains from the medial segment through an external duct that opens into the nasal cavity at the base of the vestibular fold. A duct from the lateral segment loops and opens onto the surface of the nasal septum. The structure and function of the secretory cells is reviewed briefly within the context of our study of the configuration of duck nasal salt glands.