Pattern Distribution of Connexins in the Ortho- and Parakeratinized Epithelium of the Lingual Mucosa in Birds.

Connexins are important proteins involved in cell-to-cell communication and cytodifferentiation during renewal and cornification of the multilayered epithelia. So far, there is a lack of reports on this subject in birds' structurally different ortho- and parakeratinized epithelium of the tongue. The study aims to describe the distribution and expression profiles of the α-connexins (Cx40 and 43) and ß-connexins (Cx26, 30, and 31) in those epithelia in duck, goose, and domestic turkey. Research revealed the presence of the mentioned connexins and the occurrence of interspecies differences. Connexins form gap junctions in the cell membrane or are in the cytoplasm of keratinocytes. Differences in connexin expression were noted between the basal and intermediate layers, which may determine the proliferation of keratinocytes. Cx40, 43, and Cx30 in the gap junction of the keratinocytes of the intermediate layer are related to the synchronization of the cornification process. Because of the exfoliation of cornified plaques, a lack of connexins was observed in the cornified layer of orthokeratinized epithelium. However, in parakeratinized epithelium, connexins were present in the cell membrane of keratinocytes and thus maintained cellular integrity in gradually desquamating cells. The current studies will be useful in further comparative analyses of normal and pathological epithelia of the oral cavity in birds.

Alpha-Keratin, Keratin-Associated Proteins and Transglutaminase 1 Are Present in the Ortho- and Parakeratinized Epithelium of the Avian Tongue.

The lingual mucosa in birds is covered with two specific types of multilayered epithelia, i.e., the para- and orthokeratinized epithelium, that differ structurally and functionally. Comprehensive information on proteins synthesized in keratinocyte during their cytodifferentiation in subsequent layers of multilayered epithelia in birds concerns only the epidermis and are missing the epithelia of the lingual mucosa. The aim of the present study was to perform an immunohistochemical (IHC) and molecular analysis (WB) of bird-specific alpha-keratin, keratin-associated proteins (KAPs), namely filaggrin and loricrin, as well as transglutaminase 1 in the para- and orthokeratinized epithelium covering the tongue in the domestic duck, goose, and turkey. The results reveal the presence of alpha-keratin and KAPs in both epithelia, which is a sign of the cornification process. In contrast to the epidermis, the main KAPs involved in the cornification process of the lingual epithelia in birds is loricrin. Stronger expression with KAPs and transglutaminase 1 in the orthokeratinized epithelium than in the parakeratinized epithelium may determine the formation of a more efficient protective mechanical barrier. The presence of alpha-keratin, KAPs, and transglutaminase 1 epitopes characteristic of epidermal cornification in both types of the lingual epithelia may prove that they are of ectodermal origin.

Three-dimensional characteristic of fungiform papillae and its taste buds in European bison (Bison bonasus), cattle (Bos taurus), and Bison bonasus hybrid.

BACKGROUND: Our recent macro- and scanning electron microscopic study of tongue conducted on domesticated cattle, wild living European bison, and Bison bonasus hybrid revealed various spatial arrangement and number of gustatory and mechanical papillae between parental species and their hybrid. Furthermore, scanning electron microscopy analysis of gustatory papillae indicated the variable distribution of fungiform papillae (Fu) over the surface of the tongue, which could be significant in differentiated taste perception during feeding in studied wild living and domesticated husbandry ruminants. To specify the detailed microstructure of Fu papillae with connective tissue cores (CTC) and intraepithelial taste buds system, the first time the three-dimensional computer-aided analysis of serial histoslides resulted in the rendering of 3D reconstructions of Fu papillae. RESULTS: The comparative analysis of 3D models Fu papillae conducted in six areas of lingual mucosa of each tongue revealed information about, microstructural diversity of Fu papillae in studied ruminants. The estimation of number and density of Fu papillae on tongues, rate of protrusion of papillae over mucosa, and a number of taste buds per papilla allowed to state the ventral surface of the lingual apex and posterolateral surfaces of the lingual torus as regions important in taste perception, as in the preselection of taken food, as well in the analysis of food during rumination, respectively. On the 3D models were observed three structural types of CTC of different distribution on the tongue in studied species. The quantitative data of the number of taste buds on Fu papillae have regional functional differences in the taste system important in feeding and veterinary practice. Moreover, our analysis determined specific features in examined hybrid and showed similarities of some studied features with cattle, i.e., maternal species. CONCLUSIONS: The 3D reconstruction method used for the first time in the field of study of the lingual papillae and taste buds system can be considered as an innovative and effective tool in assessing of the microstructures of Fu papillae, and it could be suitable for further studies of taste system structures in normal and pathological condition.

Unique pattern of histogenesis of the parakeratinized epithelium on lingual prominence in the domestic goose embryos (Anser anser f. domestica).

A triangular lingual prominence (LP) is a characteristic part of the tongue in Anseriformes containing adipose tissue. The parakeratinized epithelium (PEp) covers the LP. Studies aimed to describe the histogenesis of PEp during the process of the intensive formation of the LP in domestic goose during embryonic period and to determine the structural readiness to perform a protective function. The study were conducted by using LM, SEM and TEM technique. The results revealed that on day 16th the undifferentiated epithelium of LP transformed into the typical avian multilayered epithelium. Contrary to pattern of histogenesis of parakeratinized epithelium on the lingual body, on the medial and lateral areas of the elongating and bulging LP were formed epithelial furrows. Which around 20th day, on lateral areas of LP deepened up to half of epithelium, whereas on the medial area began to fade. The ultrastructure of cells lying in furrows indicated progressive apoptosis-like degeneration. On the 25th day, shallow furrows were only present on lateral areas, where bulging of LP was continued. Whereas the epithelium on medial area started cornification by the accumulation of cytokeratin fibers. Lack of the periderm during the development of the PEp of the LP indicated its endodermal origin.

Functional morphology of the tongue in the domestic turkey (Meleagris gallopavo gallopavo var. domesticus).

Nowadays, microstructural and ultrastructural analysis of organs of the avian beak cavity points to new aspects of adaptation to food intake through the various feeding groups. These data should undoubtedly be considered in the time of mass production of compound feed in poultry, when many studies analyze the optimal size of food particles and their doses. Galliformes possess complex mechanisms of food collection and transport in the beak cavity. They collect food by pecking and transport food by using catch-and-throw and slide-and-glue mechanisms. The aim of current research is to conduct functional analysis of the tongue in poultry such as domestic turkey in context of type of food, method of food intake, and transport to the esophagus. The study involves observations of macroscopic and microscopic structures of the tongue mucosa by light microscopy and scanning electron microscopy techniques with histochemical analysis of lingual glands. The obtained results showed that the tongue in domestic turkey fills two-thirds of the beak cavity. The lingual structure responsible for pecking is a rigid plate called lingual nail that works similar to a shovel to collect food. The median groove presented on surface of the tongue indicated path of food transport. The conical papillae on border between the lingual body and root are responsible for the last stage of food transport, while the papillae on the sides of root stabilize the path of food transport. For the first time, the presence of 2 types of cornified mucosal epithelia, orthokeratinized and parakeratinized epithelium, was presented. The analysis of occurrence of complex tubular lingual glands indicates production of mucous secretions composed of neutral mucopolysaccharides, with addition of sialomucins and sulfomucins. Mucous secretions moisturize surface of the tongue, thus facilitating the transport of dry food. The presence of sulfur mucopolysaccharides responds to protective function. To sum up, the tongue in domestic turkey is adapted to collect fine or coarse ground feed in form of mash or pellets through pecking and its transport to the esophagus using the slide-and-glue and throw-and-catch mechanisms.

Alpha-keratin and corneous beta protein in the parakeratinized epithelium of the tongue in the domestic goose (Anser anser f. domestica).

The parakeratinized epithelium is a common epithelium in the oral cavity in birds and is characterized by the presence of cell nuclei in the cells of the cornified layer. This epithelium covers almost the entire dorsal surface of the tongue in the domestic goose apart of the lingual nail and conical papillae. So far no study has identified the molecular proteins alpha-keratin (IF-keratin) and/or corneous beta protein (CBP), which are responsible for keratinization or cornification processes in the parakeratinized epithelium of domestic geese. The study was performed using immunohistochemical (IHC) methods to identify alpha-keratin. The innovative method of Raman microspectroscopy was used to determine the presence of CBP and specify their percentage in epithelial layers of the parakeratinized epithelium. The results revealed that alpha-keratin is present in the whole parakeratinized epithelium. A strong staining reaction was detected in the basal and intermediate layers and a less strong staining reaction in the cornified layer. Raman microspectroscopy analysis confirmed the presence of alpha-keratin and demonstrated that its percentage decreases from the basal layer to the cornified layer. The Raman microspectroscopy technique revealed the occurrence of CBP in the parakeratinized epithelium and demonstrated that the percentage of this protein increases from the basal layer to the cornified layer. Performed analysis determines that parakeratinized epithelium undergoes cornification. However, the lower percentage of CBP in the cornified layer of parakeratinized epithelium than in orthokeratinized epithelium points to the fact that parakeratinized epithelium has a weaker protective function.

Embryonic development of parakeratinized epithelium of the tongue in the domestic duck (Anas platyrhynchos f. domestica): LM, SEM, and TEM observations.

The parakeratinized epithelium is a common and widespread type of keratinized epithelium in the oral cavity in adult birds. In contrast to orthokeratinized epithelium, which mostly covers mechanical papillae and the lingual nail, parakeratinized epithelium covers almost the entire dorsal surface of the tongue in birds. The characteristic feature of parakeratinized epithelium is the presence of nuclei in the keratinized layer. The present study aimed to investigate for the first time the micro- and ultrastructural changes of parakeratinized epithelium during embryonic development and to assess the readiness of the epithelium to serve protective functions during food transport to the esophagus. Three developmental stages were distinguished: embryonic, transformation, and pre-hatching stages. The embryonic stage lasts from the 9th to the 14th day of incubation and the epithelium is composed of undifferentiated epithelial cells. The transformation stage lasts from the 15th to the 22nd day of incubation and the epithelium undergoes transformation into stratified epithelium consisting of basal, intermediate, and superficial layers. The characteristic feature of this stage is formation of the periderm with osmophilic granules. The pre-hatching stage starts on the 23rd day, and the epithelium with a fully developed keratinized layer resembles that of the epithelium in adult animals. No periderm was observed on the epithelial surface. It was confirmed that at the time of hatching the parakeratinized epithelium is fully differentiated and ready to fulfill its function during food transport. The presence of periderm is a common feature characteristic for para- and orthokeratinized epithelium in the oral cavity of birds. However, the formation of the keratinized/cornified layer is different for these two types of keratinized epithelia.

Morphology and topography of internal reproductive organs in the female cat during prenatal and postnatal development: Scanning electron microscope and three-dimensional reconstruction study.

The study describes the morphology and topography of internal reproductive organs in the domestic cat from the early prenatal period to maturity, using macroscopic and scanning electron microscope (SEM) observations with three-dimensional (3D) reconstructions. Fifty-seven female cat fetuses aged between the 27th and 63rd day postconception (p.c.), two newborn cats, three juveniles (3-month-old) cats, and three mature (12-month-old) cats were used in the study. The age of fetuses was determined on the basis of the growth curve for the domestic cat. The rudiments of cat ovaries develop on the ventral surface of the mesonephroi and within 30 days p.c. move to the sides of the abdominal cavity, which is similar to the position of the ovaries in the adult cat. The mesonephroi regress at about the 50th day p.c., when the residual mesonephric ducts are still found in the lower part of the body of the uterus. The paramesonephric ducts develop on the lateral surface of the mesonephroi and by the 45th day p.c., differentiate into the uterine tubes and the uterus. The arrangement of the paramesonephric ducts in the abdominal cavity changes from the U- to the V-shaped system. The final topography of the uterine tubes is established between the 54th and 60th day p.c., as the uterine tubes become convoluted. Before the 54th day p.c., the uterine horns undergo rapid elongation and convolution, forming the W-shaped system. By the third month of postnatal life, the uterine horns become straight, as in the adult cat.

Spexin: A novel regulator of adipogenesis and fat tissue metabolism.

Spexin (SPX, NPQ) is a novel peptide involved in the regulation of energy metabolism. SPX inhibits food intake and reduces body weight. In obese humans, SPX is the most down-regulated gene in fat. Therefore, SPX might be involved in the regulation of lipid metabolism. Here, we study the effects of SPX on lipolysis, lipogenesis, glucose uptake, adipogenesis, cell proliferation and survival in isolated human adipocytes or murine 3T3-L1 cells. SPX and its receptors, GALR2 and GALR3, are present at mRNA and protein levels in murine 3T3-L1 cells and human adipocytes. SPX inhibits adipogenesis and down-regulates mRNA expression of proadipogenic genes such as Pparγ, C/ebpα, C/ebpß and Fabp4. SPX stimulates lipolysis by increasing the phosphorylation of hormone sensitive lipase (HSL). Simultaneously, SPX inhibits lipogenesis and glucose uptake in human adipocytes and murine 3T3-L1 cells. SPX has no effect on murine 3T3-L1 cell proliferation and viability. Moreover, our research showed that the SPX effect on adipocytes metabolism is mediated via GALR2 and GALR3 receptors. SPX is a novel regulator of lipid metabolism in murine 3T3-L1 and human adipocytes.

Ultrastructural study on the embryonic development of the orthokeratinized epithelium and its cornified layer (lingual nail) on the ventral surface of the lingual apex in the domestic duck (Anas platyrhynchos f. domestica).

The lingual nail as the cornified layer of the orthokeratinized epithelium in birds is responsible for the collection of solid food by pecking. The aim of the present study is to determine the manner of orthokeratinized epithelium development and assess the degree of readiness of the epithelium to fulfill its mechanical function at hatching. Three developmental phases are distinguished, i.e. embryonic, transformation and pre-hatching stage. In the embryonic stage lasting until day 13 of incubation the epithelium is composed of several layers of undifferentiated cells. During the transformation stage, from day 14 to 20 of incubation, the epithelium becomes differentiated to form three layers. A characteristic feature is the formation of osmophilic granules in the superficial layer, referred to as periderm granules. Until the pre-hatching stage the fibrous cytoskeleton of epithelial cells and an impermeable epithelial barrier are gradually developed. In the pre-hatching stage, a cornified lingual nail is formed, while the periderm is exfoliated. At hatching the orthokeratinized epithelium and lingual nail are fully developed and ready to perform feeding activities. The presence of periderm, similarly as in the epidermis, indicates the ectodermal derivation of the oral cavity epithelium. Moreover, occurrence of osmophilic granules may be considered as evidence for the phylogenetic affinity of birds and reptiles.

The development of lingual glands in the domestic duck (Anas platyrhynchos f. domestica): 3D-reconstruction, LM, and SEM study.

The major salivary glands of birds develop by branching or elongation of the epithelial cords. The development of the minor salivary glands in form of the lingual glands has never been described. Among birds, only Anatidae have three types of the lingual glands: rostral, caudo-lateral, and caudo-medial lingual glands. The study aims to characterize the manner and rate of the lingual glands development in the domestic duck and their topographical arrangement relative to the hyoid apparatus. The study reveals that all three types of the lingual glands develop by branching. We describe five stages of the lingual glands development in the domestic ducks: prebud, initial bud, pseudoglandular, canalicular, and terminal bud stage. The pattern of the lingual glands development in birds is similar to that described for mammals, with the exception, that the terminal buds are formed at the same time as the lumen of the glands. Generally, the rostral lingual gland starts to branch earlier than the caudal lingual glands. The 3D-reconstruction shows the location and direction of lingual gland development relative to the entoglossal cartilage and basibranchial bone. Light microscopy and scanning electron microscopy allow to characterize the histogenesis of the embryonic epithelium into glandular epithelium. At a time of hatching only secretory units of caudal lingual glands resemble the secretory units of the adult domestic duck. The rostral and caudo-lateral lingual glands are arranged on the sides of the entoglossal cartilage and basibranchial bone and caudo-madial lingual glands are located over the basibranchial bone. We suggest that such an arrangement of the lingual glands in the domestic duck is important during food intake and responsible for reduction of friction and formation of food bites.

Localization of Alpha-Keratin and Beta-Keratin (Corneous Beta Protein) in the Epithelium on the Ventral Surface of the Lingual Apex and Its Lingual Nail in the Domestic Goose (Anser Anser f. domestica) by Using Immunohistochemistry and Raman Microspectroscopy Analysis.

The epithelium of the ventral surface of the apex of the tongue in most birds is specified by the presence of the special superficial layer called lingual nail. The aim of the present study is to determine the localization of the alpha-keratin and beta-keratin (corneous beta protein) in this special epithelium in the domestic goose by using immunohistochemistry staining and the Raman spectroscopy analysis. Due to lack of commercially available antibodies to detect beta-keratin (corneous beta protein), the Raman spectroscopy was used as a specific tool to detect and describe the secondary structure of proteins. The immunohistochemical (IHC) detections reveal the presence of alpha-keratin in all layers of the epithelium, but significant differences in the distribution of the alpha-keratin in the epithelial layers appear. The staining reaction is stronger from the basal layer to the upper zone of the intermediate layer. The unique result is weak staining for the alpha-keratin in the lingual nail. Applications of the Raman spectroscopy as a complementary method not only confirmed results of IHC staining for alpha-keratin, but showed that this technique could be used to demonstrate the presence of beta-keratin (corneous beta protein). Functionally, the localization of alpha-keratin in the epithelium of the ventral surface of the lingual apex provides a proper scaffold for epithelial cells and promotes structural integrity, whereas the presence of beta-keratin (corneous beta protein) in the lingual nail, described also as exoskeleton of the ventral surface of the apex, endures mechanical stress. Anat Rec, 300:1361-1368, 2017. © 2017 Wiley Periodicals, Inc.

Development of mechanical papillae of the tongue in the domestic goose (Anser anser f. domestica) during the embryonic period.

Three types of mechanical papillae, i.e., conical, filiform, and hair-like papillae, are present on the tongue in the domestic goose. Within conical papillae, we distinguish three categories: large and small conical papillae on the body and conical papillae on the lingual prominence. The arrangement of mechanical papillae on the tongue in Anseriformes is connected functionally with different feeding mechanisms such as grazing and filter-feeding. The present work aims to determine whether morphology of three types of mechanical papillae in goose at the time of hatching is the same as in an adult bird and if the tongue is prepared to fulfill feeding function. Our results revealed that the primordia of the large conical papillae start to develop during the differentiation stage. The primordia of the small conical papillae and conical papillae of the lingual papillae start to develop during the growth stage. At the end of the growth stage, only large conical papillae, three pairs of small conical papillae, and conical papillae of the lingual prominence have similar arrangement as in an adult bird. The shape and arrangement of the remaining small conical papillae probably will be changed after hatching. During embryonic period, the filiform papillae and hair-like papillae are not formed. The embryonic epithelium that covered the mechanical papillae undergoes transformation leading to the formation of multilayered epithelium. During prehatching stage, epithelium becomes orthokeratinized epithelium. In conclusion, the tongue of the domestic goose after hatching is well prepared only for grazing. The filtration of food from water is limited due to the lack of filiform papillae.

Morphofunctional study of the tongue in the domestic duck (Anas platyrhynchos f. domestica, Anatidae): LM and SEM study.

The domestic duck, as a representative of birds living in the water, is considered as a specialist filter-feeder. Behavioral observations of foraging revealed that these birds also use a terrestrial feeding mechanism such as grazing and pecking. This study examined the entirety of the lingual mucosa in relation to the structural adaptations required for this range of feeding activities. The structures on the lateral surfaces of the tongue, the conical and filiform papillae, constitute the food filtration apparatus. The process of pecking involves the spatula-shaped apex of the tongue and a specific horny plate-the lingual nail. In the grazing mechanism, large conical papillae and lamellae in the beak are required. Structures engaged in intra-oral transport include the median groove, lingual combs, the rostral border of the lingual prominence and distinct rows of conical papillae on the lingual prominence. Two types of keratinized epithelia, the ortho- and parakeratinized epithelium, as well as nonkeratinized epithelium cover individual areas of the tongue. The rostral and caudal lingual glands present in the lamina propria of the body, lingual prominence and root of the tongue produce mucus. The specific arrangement of Grandry and Herbst corpuscles form so-called bill-tongue organ monitoring food transportation. Our research confirm that the lingual mucosa in domestic duck is characterized by microstructural species-specific modifications of particular areas of the tongue, which is formed not only under the influence of the filtering mechanism, but also by terrestrial feeding mechanisms such as grazing or pecking.

LM and TEM study of the orthokeratinized and parakeratinized epithelium of the tongue in the domestic duck (Anas platyrhynchos f. domestica).

The previous histological studies of the lingual mucosa in birds characterized two types of keratinized epithelium, i.e. orthokeratinized and parakeratinized. These epithelia are composed of three layers: basal, intermediate and keratinized. The present study showed detailed ultrastructural features of cells in particular layers of two types of keratinized epithelia on the tongue in the domestic duck and defined structural differences. TEM observations showed a gradual reduction in cell organelles in the following layers, at increasing amounts of keratin fibers. The characteristic feature of the ortho- and parakeratinized epithelium is the presence of sub-layers in the intermediate layer, i.e. the upper and lower part, which results from the different shape of cell nuclei and dye affinity of the cytoplasm. The keratinized layer of ortho- and parakeratinized epithelium is built of two types of cells such as electron dark and light cells, which undergo exfoliation. The basic difference between the keratinized epithelia is the presence of flattened cell nuclei in the keratinized layer of the parakeratinized epithelium. The differentiating feature is also an arrangement of keratin fibers in the cell cytoplasm of the keratinized layer. The analysis of the thickness of the epithelium and the keratinized layer, indicated differences between keratinized epithelia, which result from two variants of performing protective functions, either through a thick keratinized layer or by a higher epithelium. Differences in the ultrastructure of the ortho- and parakeratinized epithelium are associated with mechanical functions of the epithelium resulting from different forces acting on the tongue during feeding activities.

Morphogenesis of the tongue mucosa in the domestic duck (Anas platyrhynchos f. domestica) during the late embryonic stages.

The tongue in domestic duck, as in other Anseriformes, is characterized by wide variety of shape and mechanical papillae and they fulfill different function during food collection. The present work aims to describe morphological features of the tongue as well as the pace formation of the mechanical papillae during embryonic period. The results may allow to answer whether the tongue in duck is ready to fulfill feeding function after hatching. The study revealed that the particular part of the tongue and the conical papillae of the body develop between 10th and 16th day of incubation, from the caudal part of the body into the rostral part of the tongue. The conical papillae of the lingual prominence in the first row are formed from 11th to 16th day and in the second row at the turn of the 15th and 16th day of incubation. These papillae developed symmetrically from the median part of the lingual prominence, to its edges. The lingual comb, which is used during transport of the food particles, is formed between 16th and 19th day. The present study indicated the morphological changes of the particular part of the tongue and the mechanical papillae were varied between the developmental stages. The morphology of the tongue in the domestic duck is fully developed before hatching and ready to collect food by pecking and grazing. However, filter-feeding mechanism is not still obvious.

The development of the tongue of the domestic goose from 9th to 25th day of incubation as seen by scanning electron microscopy.

The general development of the tongue in birds was described by Lillie (1908) in chicken. Bryk et al. (1992) also studied the tongue development in chicken and they observed development of the conical papillae of the body. Our study aims to describe the timing of the development of the tongue morphological features in the domestic goose by using SEM methods. The tongue of the domestic goose is characterized by the widest variety of shape of the particular part of the tongue and mechanical papillae. Results indicated that the formation of the apex, body, lingual prominence, and the root of the tongue take place between the 10th and 19th day of incubation. The tongue elongates rapidly between the 16th and 18th day of incubation. Simultaneously, the median groove appears on the body and the lingual prominence and elongates towards the rostral part of the tongue. The conical papillae of the tongue develop gradually. On the body, the conical papillae develop from the caudal part of the body to the rostral part and on the lingual prominence from the median part of the prominence to the lateral part. Hair-like papillae at the caudal surface of the body of the tongue remain primordial to the end of the incubation. Our studies on the morphogenesis of the tongue in the domestic goose revealed changes in shape of the particular part of the tongue and rapid pace of the formation of mechanical papillae. The tongue is completely develop before hatching and ready to collect food.

Functional morphology of the tongue in the domestic goose (Anser anser f. domestica).

Using LM and SEM methods, the study describes microstructures in particular areas of the tongue of the goose. A thick multilayered keratinized epithelium forms the "lingual nail" and covers small and giant conical papillae, whereby the first functions as an exoskeleton of the tongue apex, and the latter are arranged along the lingual and well-developed connective tissue cores, and together with the bill lamellae are involved in cutting. The row of conical papillae on the lingual prominence prevents regurgitation of transported food. In the area of the "lingual nail" and in the anterior part of the lingual prominence, Herbst corpuscles are accumulated, which allow to recognize food position. Filiform papillae, as widely distributed between the conical papillae of the body, are responsible for filtering. They can be explained as long keratinized processes of the epithelium and are devoid of connective tissue cores. During food transport, the flattened areas of the lingual body and the lingual prominence are protected by a parakeratinized epithelium, but the root is covered by a nonkeratinized epithelium. The presence of adipose tissue in the tongue probably reduces pressure during food passage, but also promotes mucus evacuation from the lingual glands, thus facilitating food transport. An entoglossal bone with a continuation as cartilage is the stable structural basis of the tongue system.

Functional morphology of the tongue in the nutcracker (Nucifraga caryocatactes).

The nutcracker Nucifraga caryocatactes belongs to a group of bird species that use their beak and tongue as tools for obtaining food, such as seeds from hard-to-reach cones or nuts from shells. The aim of the present study, carried out with a scanning electron microscope, was to define the morphological features of the tongue of the nutcracker, which seems to be adapted to its environment through specific methods of obtaining food. One of the characteristic features of the nutcracker's tongue is the unique structure of the anterior part of the tongue, which has two long and highly keratinized processes - a product of the renewable keratinized layer of the epithelium covering the ventral surface of the tongue. These dagger-like processes, which are a modified "lingual nail," take a major role in levering up and shelling seeds, which are transported over the short sulcus-shaped body of the tongue. A unique feature of the nutcracker's tongue is the groove separating the body from the root. Two rows of highly keratinized, mechanical, conical papillae are located at the junction of the body and the root. These papillae are mechanically protective elements for passing food particles in the form of seeds. Among lingual glands, only the posterior lingual glands on the root of the tongue have been observed. Their secretion agglutinates dry food before it is swallowed. Results of the present study indicate that the nutcracker's tongue is an efficient tool resembling a lever that is helpful in shelling seeds.