RESUMO
This is the first study to describe the subtypes, number and distribution of mast cells (MC) in cat tongue by histochemical and immunohistochemical methods. Six male adult felines' tongue tissue samples consist of the study's material. Samples were fixed in 10% formaldehyde. MC number and distribution in the feline tongue were assessed using toluidine blue. Also, sections taken from blocks were stained in alcian blue/safranin O (AB/SO) combined dyes to determine the MC subtypes. The Streptavidin biotin complex method using anti-chymase and anti-tryptase primary antibodies was used for immunohistochemistry. Metachromatic MCs were mainly observed in the lamina propria close to the multilayered keratinized stratified squamous epithelium. The high number of MCs in this region may be because the dorsal surface of the tongue plays an essential role in the defence system of tongue tissue and, thus, of the body as a whole. Additionally, the number of MCs stained with AB (+) (1.7 ± 0.08) in the feline tongue was statistically higher than those with SO (+) (0.18 ± 0.02). This might be interpreted as an indication that MC heterogeneity may be due not only to their staining properties but also to their localization. It is also conceivable that the high histamine content may be a factor in this. Tryptase-positive MCs were found in the loose connective tissue around blood vessels, between the glands, as solitary cells, or in groups of several cells. Chymase-positive MCs were observed more individually rather than in groups. Moreover, chymase-positive MCs were detected to be located in the filiform papillae subepithelial and in the blood vessels' immediate vicinity. Animals often lick themselves to clean themselves and promote healing. For this reason, it is very important to protect the tongue, which is in direct contact with the external environment, against foreign agents. Considering both the functional and protective properties of the tongue, we concluded that MCs may play a role in oral cavity immunity and protective effect.
Assuntos
Imuno-Histoquímica , Mastócitos , Língua , Animais , Gatos , Língua/citologia , Masculino , Imuno-Histoquímica/veterinária , Triptases/análise , Triptases/metabolismo , Quimases/metabolismo , Quimases/análiseRESUMO
Cattle egret (Bubulcus ibis) is a cosmopolitan heron species, with least concern conservation status. There are limited literatures on the anatomy of this bird, especially in relation to its sensory organs, hence we here investigated the gross morphological and histomorphometric features of its tongue. The tongues of twelve healthy juvenile cattle egrets were examined in situ for morphological appearance and gross morphometric measurements were determined ex situ. Routine histology was conducted on the tongue tissue with parameters such as epithelial and lamina propia heights, lingual muscle and entoglossal cartilage heights evaluated. Grossly, the tongue was divided into three parts name; apex, body and the root. It was arrow shaped, conforming to the shape of the beak, with a laryngeal mound bounded caudally by the pharyngeal papillae at its root. A massive entoglossal cartilage formed the core of the cranial apex, ventral body portion, and caudal aspect of the root. Histologically, the lingual mucosa possessed keratinized squamous epithelium in all its divisions, with spinous conical papillae being characteristic of the cranial apical mucosa. The body lingual mucosa possessed foliate papillae on the dorsal aspects, while filiform papillae were prominent in the ventral portions. The lingual root uniquely possessed numerous glandular ducts in its lamina propia as well as localized adipocytes. Overall, the regression analysis data showed that the body weight can be conveniently predicted from tongue parameters. This study has thus provided additional knowledge on the anatomy of the birds and the generated data could prove useful in comparative regional anatomy.
Assuntos
Aves , Língua , Aves/anatomia & histologia , Língua/anatomia & histologia , Língua/citologia , Análise de Regressão , Peso Corporal , Mucosa/citologia , AnimaisRESUMO
Little is known about the molecular mechanisms underlying drug-induced taste disorders, which can cause malnutrition and reduce quality of life. One of taste disorders is known adverse effects of bisphosphonates, which are administered as anti-osteoporotic drugs. Therefore, the present study evaluated the effects of risedronate (a bisphosphonate) on taste bud cells. Expression analyses revealed that farnesyl diphosphate synthase (FDPS, a key enzyme in the mevalonate pathway) was present in a subset of mouse taste bud and tongue epithelial cells, especially type III sour-sensitive taste cells. Other mevalonate pathway-associated molecules were also detected in mouse taste buds. Behavioral analyses revealed that mice administered risedronate exhibited a significantly enhanced aversion to HCl but not for other basic taste solutions, whereas the taste nerve responses were not affected by risedronate. Additionally, the taste buds of mice administered risedronate exhibited significantly lower mRNA expression of desmoglein-2, an integral component of desmosomes. Taken together, these findings suggest that risedronate may interact directly with FDPS to inhibit the mevalonate pathway in taste bud and tongue epithelial cells, thereby affecting the expression of desmoglein-2 related with epithelial barrier function, which may lead to alterations in behavioral responses to HCl via somatosensory nerves.
Assuntos
Difosfonatos , Células Epiteliais , Geraniltranstransferase , Animais , Camundongos , Difosfonatos/farmacologia , Células Epiteliais/enzimologia , Geraniltranstransferase/genética , Qualidade de Vida , Distúrbios do Paladar , Papilas Gustativas/citologia , Língua/citologia , Ácido Risedrônico/farmacologiaRESUMO
Taste bud cells are renewed throughout life in a process requiring innervation. Recently, we reported that R-spondin substitutes for neuronal input for taste cell regeneration. R-spondin amplifies WNT signaling by interacting with stem-cell-expressed E3 ubiquitin ligases RNF43/ZNRF3 (negative regulators of WNT signaling) and G-protein-coupled receptors LGR4/5/6 (positive regulators of WNT signaling). Therefore, we hypothesized that RNF43/ZNRF3 may serve as a brake, controlled by gustatory neuron-produced R-spondin, for regulating taste tissue homeostasis. Here, we show that mice deficient for Rnf43/Znrf3 in KRT5-expressing epithelial stem/progenitor cells (RZ dKO) exhibited taste cell hyperplasia; in stark contrast, epithelial tissue on the tongue degenerated. WNT signaling blockade substantially reversed all these effects in RZ dKO mice. Furthermore, innervation becomes dispensable for taste cell renewal in RZ dKO mice. We thus demonstrate important but distinct functions of RNF43/ZNRF3 in regulating taste versus lingual epithelial tissue homeostasis.
Assuntos
Epitélio/metabolismo , Língua/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Animais , Benzenoacetamidas/farmacologia , Nervo Glossofaríngeo/cirurgia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Piridinas/farmacologia , Células-Tronco/citologia , Células-Tronco/metabolismo , Paladar/fisiologia , Papilas Gustativas/metabolismo , Língua/citologia , Ubiquitina-Proteína Ligases/deficiência , Ubiquitina-Proteína Ligases/genética , Via de Sinalização Wnt/efeitos dos fármacosRESUMO
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.
Assuntos
Embrião não Mamífero/citologia , Epitélio/embriologia , Queratinas/metabolismo , Organogênese , Língua/citologia , Animais , Embrião não Mamífero/metabolismo , Epitélio/metabolismo , Gansos , Língua/metabolismoRESUMO
Leptin is known to selectively suppress neural and taste cell responses to sweet compounds. The sweet suppressive effect of leptin is mediated by the leptin receptor Ob-Rb, and the ATP-gated K+ (KATP ) channel expressed in some sweet-sensitive, taste receptor family 1 member 3 (T1R3)-positive taste cells. However, the intracellular transduction pathway connecting Ob-Rb to KATP channel remains unknown. Here we report that phosphoinositide 3-kinase (PI3K) mediates leptin's suppression of sweet responses in T1R3-positive taste cells. In in situ taste cell recording, systemically administrated leptin suppressed taste cell responses to sucrose in T1R3-positive taste cells. Such leptin's suppression of sucrose responses was impaired by co-administration of PI3K inhibitors (wortmannin or LY294002). In contrast, co-administration of signal transducer and activator of transcription 3 inhibitor (Stattic) or Src homology region 2 domain-containing phosphatase-2 inhibitor (SHP099) had no effect on leptin's suppression of sucrose responses, although signal transducer and activator of transcription 3 and Src homology region 2 domain-containing phosphatase-2 were expressed in T1R3-positive taste cells. In peeled tongue epithelium, phosphatidylinositol (3,4,5)-trisphosphate production and phosphorylation of AKT by leptin were immunohistochemically detected in some T1R3-positive taste cells but not in glutamate decarboxylase 67-positive taste cells. Leptin-induced phosphatidylinositol (3,4,5)-trisphosphate production was suppressed by LY294002. Thus, leptin suppresses sweet responses of T1R3-positive taste cells by activation of Ob-Rb-PI3K-KATP channel pathway.
Assuntos
Leptina/farmacologia , Fosfatidilinositol 3-Quinases/fisiologia , Receptores Acoplados a Proteínas G/efeitos dos fármacos , Edulcorantes/farmacologia , Papilas Gustativas/efeitos dos fármacos , Paladar/efeitos dos fármacos , Animais , Feminino , Masculino , Camundongos , Camundongos Transgênicos , Fosfatidilinositóis/metabolismo , Inibidores de Proteínas Quinases/farmacologia , Fator de Transcrição STAT3/genética , Transdução de Sinais/efeitos dos fármacos , Língua/citologia , Língua/efeitos dos fármacosRESUMO
Objective To investigate the expression and clinical significance of phosphatase and tension homolog deleted on chromosome ten (PTEN) and stimulator of interferon genes (STING) in human tongue squamous cell carcinoma (TSCC). Methods The expression of PTEN and STING protein in 65 pairs of TSCC and paracancerous tissues was detected by immunohistochemical EnVision method, and the relationships between PTEN, STING and clinicopathological parameters, overall survival (OS) and prognosis were analyzed by statistical methods. Results Compared with the adjacent tissues, the expression of PTEN in TSCC significantly decreased, and the expression of STING significantly increased. PTEN was negatively correlated with STING. In TSCC, the expression of PTEN and STING were correlated with pathological grade, TNM stage and lymph node metastasis. There were no significant correlations between the expression intensity of PTEN, STING and the gender and age of patients. The low expression of PTEN and the high expression of STING in TSCC tissues were significantly associated with poor prognosis and significantly shortened overall survival of patients. Conclusion TSCC patients with low expression of PTEN and high expression of STING have poor prognosis and short survival time. Combined detection of PTEN and STING expression is helpful to evaluate the degree of tumor progression and patient prognosis.
Assuntos
Carcinoma de Células Escamosas , Proteínas de Membrana , PTEN Fosfo-Hidrolase , Neoplasias da Língua , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/genética , Humanos , Proteínas de Membrana/genética , PTEN Fosfo-Hidrolase/genética , Prognóstico , Língua/citologia , Neoplasias da Língua/diagnóstico , Neoplasias da Língua/genéticaRESUMO
Taste bud cells arise from local epithelial stem cells in the oral cavity and are continuously replaced by newborn cells throughout an animal's life. However, little is known about the molecular and cellular mechanisms of taste cell turnover. Recently, it has been demonstrated that SOX2, a transcription factor expressed in epithelial stem/progenitor cells of the oral cavity, regulates turnover of anterior tongue epithelium including gustatory and non-gustatory papillae. Yet, the role of SOX2 in regulating taste cell turnover in the posterior tongue is unclear. Prompted by the fact that there are regional differences in the cellular and molecular composition of taste buds and stem/progenitor cells in the anterior and posterior portions of tongue, which are derived from distinct embryonic origins, we set out to determine the role of SOX2 in epithelial tissue homeostasis in the posterior tongue. Here we report the differential requirement of SOX2 in the stem/progenitor cells for the normal turnover of lingual epithelial cells in the posterior tongue. Sox2 deletion in the stem/progenitor cells neither induced active caspase 3-mediated apoptotic cell death nor altered stem/progenitor cell population in the posterior tongue. Nevertheless, morphology and molecular feature of non-gustatory epithelial cells were impaired in the circumvallate papilla but not in the filiform papillae. Remarkably, taste buds became thinner, collapsed, and undetectable over time. Lineage tracing of Sox2-deleted stem/progenitor cells demonstrated an almost complete lack of newly generated basal precursor cells in the taste buds, suggesting mechanistically that Sox2 is involved in determining stem/progenitor cells to differentiate to gustatory lineage cells. Together, these results demonstrate that SOX2 plays key roles in regulating epithelial tissue homeostasis in the posterior tongue, similar but not identical to its function in the anterior tongue.
Assuntos
Epitélio/metabolismo , Fatores de Transcrição SOXB1/metabolismo , Papilas Gustativas/metabolismo , Língua , Animais , Diferenciação Celular , Homeostase , Hibridização In Situ , Camundongos , Células-Tronco/metabolismo , Língua/citologia , Língua/metabolismoRESUMO
BACKGROUND: The herbicide dichlorophenoxyacetic acid (2,4-D) is one of the most widely used crop spraying products in the world. Some pesticides induce the degranulation of mast cells and increase allergic responses. This is the first study to evaluate the damage to the oral mucosa after an experimental simulation of environmental inhalation exposure to the 2,4-D herbicide. The aim of this study was evaluate the possible oral damage caused by acute inhalation exposure to the herbicide 2,4-D. RESULTS: There was a difference between the exposure concentrations in relation to tissue congestion intensity (p = 0.002) and mast cell counts (p = 0.002), a difference in the evaluation of the interaction between the exposure concentrations and nebulization time in the dorsum epithelium thickness (p = 0.013), and a significant correlation between the epithelial thickness and the number of nucleoli organizing regions on the dorsum of the tongue (p = 0.048). CONCLUSIONS: Even after acute exposure, the herbicide 2,4-D had the potential to damage the oral epithelium, especially at higher doses.
Assuntos
Ácido 2,4-Diclorofenoxiacético/toxicidade , Herbicidas/toxicidade , Exposição por Inalação/efeitos adversos , Mucosa Bucal/patologia , Animais , Epitélio/patologia , Masculino , Mastócitos , Camundongos , Mucosa Bucal/citologia , Região Organizadora do Nucléolo , Língua/citologia , Língua/patologiaRESUMO
Sense of taste is central to evaluate food before digestion. Taste stem cells undergo constant differentiation throughout the life. However, the mechanism underlying the generation of taste receptor cells is still not clear. Here, we cultured taste organoids from either Lgr5+ or Lgr5-cells, and found the preferential generation of Car4+ and Gustducin + taste receptor cells in organoids derived from Lgr5+ cells in circumvallate, foliate or fungiform papillae. Taste organoids derived from Lgr5+ cells in circumvallate papillae of neonatal mice showed stronger capacity to generate taste receptor cells compared to the organoids from Lgr5+ cells of the adult circumvallate papillae. Massive transcriptional differences were found in multiple signaling pathways including taste transduction between organoids derived from circumvallate papillae of adult and neonatal mice. Inhibiting the Notch signaling pathway by LY411575 enhanced taste receptor cell generation in organoids from circumvallate papillae and modulated multiple signaling pathways. Thus, we concluded that receptor cell generation in taste organoids was age-related and regulated via multiple signaling pathways.
Assuntos
Envelhecimento/fisiologia , Organoides/citologia , Organoides/metabolismo , Transdução de Sinais , Papilas Gustativas/citologia , Alanina/análogos & derivados , Alanina/farmacologia , Animais , Animais Recém-Nascidos , Azepinas/farmacologia , Caderinas/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Organoides/efeitos dos fármacos , RNA-Seq , Receptores Acoplados a Proteínas G/metabolismo , Receptores Notch/metabolismo , Paladar , Língua/citologia , Transcrição Gênica/efeitos dos fármacosRESUMO
Mesenchymal stem cells (MSCs) possess the ability to differentiate into multiple cell lineages, and thus, confer great potential for use in regenerative medicine and biotechnology. In the present study, we attempted to isolate and characterize bovine tongue tissue epithelium-derived MSCs (boT-MSCs) and investigate the culture conditions required for long-term culturing of boT-MSCs. boT-MSCs were successfully isolated by the collagenase digestion method and their proliferative capacity was maintained for up to 20 or more passages. We observed a significant increase in the proliferation of boT-MSCs during the 20 consecutive passages under low-glucose Dulbecco's modified Eagle's medium culture condition among the three culture conditions. These boT-MSCs presented pluripotency markers (octamer-binding transcription factor 3/4 (Oct3/4) and sex determining region Y-box2 (Sox2)) and cell surface markers, which included CD13, CD29, CD44, CD73, CD90, CD105, CD166, and major histocompatibility complex (MHC) class I (MHC-I) but not CD11b, CD14, CD31, CD34, CD45, CD80, CD86, CD106, CD117, and MHC-II at third passage. Moreover, these boT-MSCs could differentiate into mesodermal (adipocyte, osteocyte, and chondrocyte) cell lineages. Thus, the present study suggests that the tongue of bovines could be used as a source of bovine MSCs.
Assuntos
Células Epiteliais/fisiologia , Células-Tronco Mesenquimais/fisiologia , Mucosa Bucal/citologia , Língua/citologia , Animais , Bovinos , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Cultura Primária de CélulasRESUMO
Extraocular muscles (EOMs) show resistance to muscle dystrophies and sarcopenia. It has been recently demonstrated that they are endowed with different types of myogenic cells, all of which present an outstanding regenerative potential. Neurotrophins are important modulators of myogenic regeneration and act promoting myoblast proliferation, enhancing myogenic fusion rates and protecting myotubes from inflammatory stimuli. Here, we adapted the pre-plate cell isolation technique to obtain myogenic progenitors from the rat EOMs, and quantified their in vitro expression of neurotrophins and their receptors by RT-qPCR and immunohistochemistry, respectively. The results were compared with the expression on progenitors isolated from buccinator, tongue and limb muscles. Our quantitative analysis of brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF) and neurotrophin-3 (NT-3) transcripts showed, for the first time, that EOMs-derived cells express more of these factors and that they expressed TrkA, but not TrkB and TrkC receptors. On the contrary, the immunofluorescence analysis demonstrated high expression of p75NTR on all myogenic progenitors, with the EOMs-derived cells showing higher expression. Taken together, these results suggest that the intrinsic trophic differences between EOMs-derived myogenic progenitors and their counterparts from other muscles could explain why those cells show higher proliferative and fusion rates, as well as better regenerative properties.
Assuntos
Músculo Esquelético/citologia , Fatores de Crescimento Neural/metabolismo , Músculos Oculomotores/metabolismo , Receptores de Fator de Crescimento Neural/metabolismo , Células-Tronco/metabolismo , Animais , Contagem de Células , Diferenciação Celular , Células Cultivadas , Desenvolvimento Muscular , Ratos Wistar , Língua/citologiaRESUMO
The tayra (Eira barbara) is a mammal belonging to the Mustelidae family that occurs in all Brazilian biomes. The present work aimed to describe the morphology of the tongue of these specimens highlighting their structures and particularities that will serve as a subsidy to elucidate the anatomy of the same and for comparative studies among other species of domestic and wild animals. Five adult male specimens of E. barbara were studied, which were fixed using 10% aqueous formaldehyde solution. The tongue was removed by opening the oral cavity and separating the maxillary/mandible bone complex. Being in possession of the material, photodocumentations and collection of the fragments were made for the proper preparation of histological slides and Scanning Electron Microscopy (SEM). The lingual papillae found in tayra were mechanical: filiform and conical; and gustative: fungiform and circumvallated. Histologically, the papillae are constituted by keratinized stratified epithelium and in the innermost region, it was composed of tissue connective dense unshaped followed by a layer of muscle bundles of skeletal striated. In the region of the root of the tongue of E. barbara, there were a set of small mixed salivary glands (serous and mucous) and the punctual presence of gustatory corpuscles at the level of epithelium. The morphological description of the E. barbara tongue revealed similarity to that described in literature for other domestic and wild mammals. However, the particularity of the absence of foliate papilla and the quantitative of four papillae circumvallate in the region of the root of the tongue of this species.
Assuntos
Mustelidae , Papilas Gustativas/ultraestrutura , Língua , Animais , Animais Selvagens/anatomia & histologia , Microscopia Eletrônica de Varredura/veterinária , Mustelidae/anatomia & histologia , Papilas Gustativas/citologia , Língua/anatomia & histologia , Língua/citologia , Língua/ultraestruturaRESUMO
Bridging the gap between two-dimensional cell cultures and complex in vivo tissues, three-dimensional cell culture models are of increasing interest in the fields of cell biology and pharmacology. However, present challenges hamper live cell imaging of three-dimensional cell cultures. These include (i) the stabilization of these structures under perfusion conditions, (ii) the recording of many z-planes at high spatio-temporal resolution, (iii) and the data analysis that ranges in complexity from whole specimens to single cells. Here, we addressed these issues for the time-lapse analysis of Ca2+ signaling in spheroids composed of human tongue-derived HTC-8 cells upon perfusion of gustatory substances. Live cell imaging setups for confocal and light sheet microscopy were developed that allow simple and robust spheroid stabilization and high-resolution microscopy with perfusion. Visualization of spheroids made of HTC-8 cells expressing the G-GECO fluorescent Ca2+ sensor revealed Ca2+ transients that showed similar kinetics but different amplitudes upon perfusion of bitter compounds Salicine and Saccharin. Dose-dependent responses to Saccharin required extracellular Ca2+. From the border towards the center of spheroids, compound-induced Ca2+ signals were progressively delayed and decreased in amplitude. Stimulation with ATP led to strong Ca2+ transients that were faster than those evoked by the bitter compounds and blockade of purinergic receptors with Suramin abutted the response to Saccharin, suggesting that ATP mediates a positive autocrine and paracrine feedback. Imaging of ATP-induced Ca2+ transients with light sheet microscopy allowed acquisition over a z-depth of 100 µm without losing spatial and temporal resolution. In summary, the presented approaches permit the study of fast cellular signaling in three-dimensional cultures upon compound perfusion.
Assuntos
Sinalização do Cálcio , Técnicas de Cultura de Células , Imageamento Tridimensional , Perfusão , Sacarina/farmacologia , Língua/citologia , Trifosfato de Adenosina/metabolismo , Cálcio/metabolismo , Sinalização do Cálcio/efeitos dos fármacos , Linhagem Celular , Difusão , Humanos , Rodaminas/metabolismo , Transdução de Sinais/efeitos dos fármacos , Esferoides Celulares/citologia , Esferoides Celulares/efeitos dos fármacosRESUMO
This study was conducted with the tongue samples of different life stages of hilsa, that is, adult Marine hilsa, adult Riverine hilsa, and Riverine juvenile hilsa, respectively. Three types of taste buds (Types I, II, and III based on their elevation from the epithelium at different levels) of the tongue, which may be to ensure full utilization of the gustatory ability of the fish were rocorded. Presence of specific taste buds indicate that the fish hilsa dwells in turbid waters with a possible preference toward diatom like planktonic food source. Enhanced expression of taste receptors (T1R1 and T1R3) and associated stimulatory G-proteins subunits on tongue also indicate occurrence of amino acid like substances that guided sensory cues for feeding by this fish. A firm regularity or stringency of the free surface of the epithelial cells may be attributed to compactly arranged microridges. These structures protect against physical abrasions potentially caused during food manoeuvring and swallowing. In our present observations, the surface architectures of the tongue of hilsa are discussed within the background of migratory adaptation of the species in the context of feeding and habitat preferences.
Assuntos
Peixes/anatomia & histologia , Papilas Gustativas/ultraestrutura , Língua/ultraestrutura , Animais , Estágios do Ciclo de Vida , Papilas Gustativas/citologia , Língua/citologiaRESUMO
During a foot-and-mouth disease (FMD) outbreak, transport and testing of potentially infectious samples, including epithelium from suspect lesions, presents a biosafety risk, particularly in FMD-free countries. Therefore, treatment to inactivate virus prior to transport is important. Tongue epithelium from cattle infected with FMD virus (FMDV) serotype O (O ALG/3/2014 - Lineage O/ME-SA/Ind-2001d) or A (A IRN/22/2015 - Lineage A/ASIA/G-VII) was incubated in RNAlater, RNA Shield or phosphate-buffered saline (pH 7.4) at room temperature for 2, 6, 24 or 48 h. After incubation, tissues were homogenised and tested by virus titration. Viral RNA in the homogenate was quantified by RT-qPCR, used for sequencing, and transfected into LFBKαVß6 cells to recover infectious virus. RNAlater reduced A IRN/22/2015 titres by 4 log10 after 24 h, and completely after 48 h incubation. While O ALG/3/2014 was detected by VI after 2, 6 and 24 h, titration yielded no infectious virus, likely as a result of freeze-thawing. RNA Shield was cytotoxic at high concentrations but was effective at inactivating both strains after 24 h. Regardless of reagent or inactivation period, RT-qPCR, VP1 sequencing, and transfection of RNA to recover infectious virus were possible. RNA Shield appears a better choice for FMDV inactivation in tissues, however 24 h incubation is recommended.
Assuntos
Epitélio/virologia , Vírus da Febre Aftosa/fisiologia , Manejo de Espécimes/métodos , Inativação de Vírus , Animais , Bovinos , Contenção de Riscos Biológicos , Vírus da Febre Aftosa/efeitos dos fármacos , Língua/citologia , Língua/virologia , Meios de TransporteRESUMO
In this study, we aimed to observe the localization and expression of peptide hormones-leptin, ghrelin and obestatin-in the sheep tongue by immunohistochemistry. For that purpose, tongues of ten adult sheep were used. Leptin expression of moderate intensity was observed in the basal and parabasal epithelial cells of the luminal epithelium, and leptin was strongly expressed in the taste buds of the circumvallate and fungiform papillae and in von Ebner's glands. Ghrelin was primarily expressed in some of the skeletal muscle cells and the smooth muscle cells of the middle layer of blood vessels. A strong expression was observed in the epithelial cells lining the base of the groove surrounding the circumvallate papillae. Obestatin expression was particularly strong in the epithelial cells of the salivary ducts. It was also stronger in the von Ebner's glands than in the seromucous glands. Leptin, ghrelin and obestatin were shown to be produced at varying levels in different cell types, including epithelial, stromal and skeletal muscle cells, as well as in ganglion neurons, neural plexuses and blood vessels in the sheep tongue. Cellular localization and expression of these peptide hormones have not been investigated in many species including sheep.
Assuntos
Hormônios Peptídicos/metabolismo , Língua , Animais , Vasos Sanguíneos/metabolismo , Células Epiteliais/metabolismo , Grelina/metabolismo , Imuno-Histoquímica , Leptina/metabolismo , Células Musculares/metabolismo , Neurônios/metabolismo , Receptores de Grelina/metabolismo , Ovinos , Papilas Gustativas/metabolismo , Língua/citologia , Língua/metabolismoRESUMO
Bumblebees and some other tiny animals feed on nectar by visiting flowers in their neighborhood. Some bee species appear to be highly specialized, their tongue being adapted to specific flowers. Bombus terrestris in contrast is able to feed on a wide variety of flowers and can thus be considered as a kind of universal nectar catcher. Since plant nectars show highly variable sugar content, Bombus terrestris have developed a capture mechanism that works for almost any fluid viscosity. Their tongues are decorated with very elongated papillae forming a hairy coating surrounding a rod-like main stalk. When settled on a flower, Bombus rapidly dip their tongue into the inflorescence to catch the highly sought-after nectar. To determine the physical mechanism at the origin of this outstanding ability, the capture dynamics was followed from videos recorded during viscous fluid ingestion. Surprisingly, the volume per lap and the lapping frequency are independent of the fluid viscosity over three orders of magnitude. To explain this observation, we designed a physical model of viscous dipping with structured rods. Predictions of the model compared to observations for bees showed that the nectar is not captured with the help of viscous drag, as proposed in the Landau-Levich-Derjaguin model, but thanks to the hairy structure that traps the viscous fluid, capillary forces drastically limiting the drainage. Our approach can be transposed to others nectar foragers such as bats and hummingbirds.
Assuntos
Abelhas , Líquidos Corporais/metabolismo , Flores/química , Néctar de Plantas/metabolismo , Língua/metabolismo , Animais , Comportamento Animal , Fenômenos Biomecânicos , Carboidratos/química , Modelos Biológicos , Língua/citologia , Gravação em VídeoRESUMO
Neuronatin (Nnat) is expressed in the pituitary, pancreas, and other tissues; however, the function of NNAT is still unclear. Recent studies have demonstrated that NNAT is localized in the sex-determining region Y-box 2-positive stem/progenitor cells in the developing rat pituitary primordium and is downregulated during differentiation into mature hormone-producing cells. Moreover, NNAT is widely localized in subcellular organelles, excluding the Golgi. Here, we further evaluated NNAT-positive cells and intracellular localization in embryonic and postnatal rat tissues such as the pancreas, tongue, whisker hair follicle, and testis. Immunohistochemistry revealed that NNAT was localized in undifferentiated cells (i.e., epithelial basal cells and basement cells in the papillae of the tongue and round and elongated spermatids of the testis) as well as in differentiated cells (insulin-positive cells and exocrine cells of the pancreas, taste receptor cells of the fungiform papilla, the inner root sheath of whisker hair follicles, and spermatozoa). In addition, NNAT exhibited novel intracellular localization in acrosomes in the spermatozoa. Because the endoplasmic reticulum (ER) is excluded from spermatozoa and sarco/ER Ca2+-ATPase isoform 2 (SERCA2) is absent from the inner root sheath, these findings suggested that NNAT localization in the ER and its interaction with SERCA2 are cell- or tissue-specific properties.
Assuntos
Folículo Piloso/metabolismo , Proteínas de Membrana/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Pâncreas/metabolismo , Testículo/metabolismo , Língua/metabolismo , Animais , Sinalização do Cálcio , Feminino , Glucose/metabolismo , Folículo Piloso/citologia , Espaço Intracelular/metabolismo , Masculino , Especificidade de Órgãos , Pâncreas/citologia , Gravidez , Ratos , Ratos Wistar , Testículo/citologia , Língua/citologiaRESUMO
The vertebrate tongue is a complex muscular organ situated in the oral cavity and involved in multiple functions including mastication, taste sensation, articulation and the maintenance of oral health. Although the gross embryological contributions to tongue formation have been known for many years, it is only relatively recently that the molecular pathways regulating these processes have begun to be discovered. In particular, there is now evidence that the Hedgehog, TGF-Beta, Wnt and Notch signaling pathways all play an important role in mediating appropriate signaling interactions between the epithelial, cranial neural crest and mesodermal cell populations that are required to form the tongue. In humans, a number of congenital abnormalities that affect gross morphology of the tongue have also been described, occurring in isolation or as part of a developmental syndrome, which can greatly impact on the health and well-being of affected individuals. These anomalies can range from an absence of tongue formation (aglossia) through to diminutive (microglossia), enlarged (macroglossia) or bifid tongue. Here, we present an overview of the gross anatomy and embryology of mammalian tongue development, focusing on the molecular processes underlying formation of the musculature and connective tissues within this organ. We also survey the clinical presentation of tongue anomalies seen in human populations, whilst considering their developmental and genetic etiology.