520 research outputs found

    Evidence of solitary chemosensory cells in a large mammal: the diffuse chemosensory system in Bos taurus airways

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    The diffuse chemosensory system (DCS) of the respiratory apparatus is composed of solitary chemosensory cells (SCCs) that resemble taste cells but are not organized in end organs. The discovery of the DCS may open up new approaches to respiratory diseases. However, available data on mammalian SCCs have so far been collected from rodents, the airways of which display some differences from those of large mammals. Here we investigated the presence of the DCS and of SCCs in cows and bulls (Bos taurus), in which the airway cytology is similar to that in humans, focusing our attention on detection in the airways of molecules involved in the transduction cascade of taste [i.e. alpha-gustducin and phospholipase C of the beta 2 subtype (PLC beta 2)]. The aim of the research was to extend our understanding of airway chemoreceptors and to compare the organization of the DCS in a large mammal with that in rodents. Using immunocytochemistry for alpha-gustducin, the taste buds of the tongue and arytenoid were visualized. In the trachea and bronchi, alpha-gustducin-immunoreactive SCCs were frequently found. Using immunocytochemistry for PLC beta 2, the staining pattern was generally similar to those seen for alpha-gustducin. Immunoblotting confirmed the expression of alpha-gustducin in the tongue and in all the airway regions tested. The study demonstrated the presence of SCCs in cows and bulls, suggesting that DCSs are present in many mammalian species. The description of areas with a high density of SCCs in bovine bronchi seems to indicate that the view of the DCS as made up of isolated cells totally devoid of ancillary elements is probably an oversimplification

    Neuronal intermediate filaments in the developing tongue of the frog Rana

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    The expression of several neuronal intermediate filament (NIF) proteins was investigated in the tongue of metamorphosing tadpoles (stage 38-45 of Gosner) and in adult individuals of the frog, Rana esculenta by means of immunohistochemistry. Results showed that nerve fibres at early stages of tongue development expressed peripherin (a NIF protein usually found in differentiating neurones) as well as the light and medium molecular weight NIF polypeptide subunits (NF-L and NF-M, respectively); in the adult frog, peripherin was still found in nerve fibres reaching the fungiform papilla together with NF-M, but NF-L immunoreactivity was absent therein. Clusters of epithelial cells expressing peripherin were found in the early developing tongue before differentiation of taste organs, and NF-L and NF-H immunoreactivities were present in basal (Merkel) cells of the adult frog taste disc. Results indicate that neurones innervating the adult frog's taste disc maintain a certain plasticity in their cytoskeleton and that neuronal-like cells are present in the undifferentiated and differentiated tongue epithelium possibly playing a role in the developing and mature taste organ

    Eukaryotic vs. prokaryotic chemosensory systems.

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    In the last decades, microbiologists demonstrated that microorganisms possess chemosensory capabilities and communicate with each other via chemical signals. In parallel, it was demonstrated that solitary eukaryotic chemosensory cells are diffusely located on the mucosae of digestive and respiratory apparatuses. It is now evident that on the mucosal surfaces of vertebrates, two chemoreceptorial systems (i.e. eukaryotic and prokaryotic) coexist in a common microenvironment. To date, it is not known if the two chemosensory systems reciprocally interact and compete for detection of chemical cues. This appears to be a fruitful field of study and future researches must consider that the mucosal epithelia possess more chemosensory capabilities than previously supposed

    Timing of neuronal intermediate filament proteins expression in the mouse

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    Several types of intermediate filament proteins are expressed in developing and mature neurons; they cooperate with other cytoskeletal components to sustain neuronal function from early neurogenesis onward. In this work the timing of expression of nestin, peripherin, internexin, and the neuronal intermediate filament triplet [polypeptide subunits of low (NF-L), medium (NF-M), and high (NF-H) molecular weight] was investigated in the developing fetal and postnatal mouse vomeronasal organ (VNO) by means of immunohistochemistry. The results show that the sequence of expression of intermediate filament proteins is internexin, nestin, and NF-M in the developing vomeronasal sensory epithelium; internexin, peripherin, and NF-M in the developing vomeronasal nerve; and nestin, internexin and peripherin, NF-L, and NF-M in the nerve supply to accessory structures of the VNO. At sexual maturity (2 months) NF-M is only expressed in vomeronasal neurons and NF-M, NF-L and peripherin are expressed in extrinsic nerves supplying VNO structures. The differential distribution of intermediate filament proteins in the vomeronasal sensory epithelium and nerve is discussed in terms of the cell types present therein. It is concluded that several intermediate filament proteins are sequentially expressed during intrauterine development of the VNO neural structures in a different pattern according to the different components of the VNO

    Glucose transporters are expressed in taste receptor cells.

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    In the intestine, changes of sugar concentration generated in the lumen during digestion induce adaptive responses of glucose transporters in the epithelium. A close matching between the intestinal expression of glucose transporters and the composition and amount of the diet has been provided by several experiments. Functional evidence has demonstrated that the regulation of glucose transporters into enterocytes is induced by the sensing of sugar of the enteroendocrine cells through activation of sweet taste receptors (T1R2 and T1R3) and their associated elements of G-protein-linked signaling pathways (e.g. α-gustducin, phospholipase C β type 2 and transient receptor potential channel M5), which are signaling molecules also involved in the perception of sweet substances in the taste receptor cells (TRCs) of the tongue. Considering this phenotypical similarity between the intestinal cells and TRCs, we evaluated whether the TRCs themselves possess proteins of the glucose transport mechanism. Therefore, we investigated the expression of the typical intestinal glucose transporters (i.e. GLUT2, GLUT5 and SGLT1) in rat circumvallate papillae, using immunohistochemistry, double-labeling immunofluorescence, immunoelectron microscopy and reverse transcriptase-polymerase chain reaction analysis. The results showed that GLUT2, GLUT5 and SGLT1 are expressed in TRCs; their immunoreactivity was also observed in cells that displayed staining for α-gustducin and T1R3 receptor. The immunoelectron microscopic results confirmed that GLUT2, GLUT5 and SGLT1 were predominantly expressed in cells with ultrastructural characteristics of chemoreceptor cells. The presence of glucose transporters in TRCs adds a further link between chemosensory information and cellular responses to sweet stimuli that may have important roles in glucose homeostasis, contributing to a better understanding of the pathways implicated in glucose metabolism

    Neurochemistry of the gustatory subgemmal plexus

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    Nerve fibers present in the basal plexus of the vallate papilla of the rat tongue were analyzed using cytochemical, immunocytochemical and ultrastructural methods to investigate whether the subgemmal plexus is subdivided into neurochemical compartments and to provide a clear definition of the reciprocal spatial relationships between nitrergic, peptidergic and acetylesterase positive structures. Several neuronal fibers were detected under the chemoreceptorial epithelium. Some of these fibers were in contact with the taste buds and in some cases neuronal projections were also present between the buds or inside them; some others fibers were present below this layer but in a more peripheral area. Antibodies against CGRP, SP and CCK stained fibers just below the chemoreceptorial epithelium, whereas fibers more distally located were immunolabeled by anti VIP, NOS-1 and NF-200 antibodies. Some double staining experiments were conducted using confocal microscopy. Other sections were processed cytochemically for AChE and subsequently for NADPH-d in colocalization experiments. All the data obtained using these techniques confirmed the results obtained with single immunostaining, as did the ultrastructural results. In conclusion, the present work demonstrates that the subgemmal plexus is a bilayered structure, suggesting that the complex relationship between the two layers plays a pivotal role in taste and in the control of processes ancillary to taste, such as control of vascular or secretory mechanisms

    Epithelial membrane transporters expression in the developing to adult mouse vomeronasal organ and olfactory mucosa.

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    In order to contribute clarifying mechanisms operating in nose chemosensory epithelia and their developmental patterns, we analysed the expression of different epithelial membrane transporters as well as the Clara cell secretory protein, CC26 in the olfactory, vomeronasal organ (VNO), and respiratory epithelia of embryonic (E13 to E19) and postnatal (P8 to P60) mice by means of immunohistochemistry and reverse transcriptase-polymerase chain reaction. Results showed that CC26, cAMP-activated chloride channel (CFTR), and the water channel protein aquaporin 2, 3, 4, and 5 (AQP2, AQP3, AQP4, AQP5) are expressed in developing to adult chemosensory epithelia with differential timing; moreover, their pattern of expression is not identical in VNO and olfactory epithelia as well as the corresponding associated glands; co-localization experiments using olfactory marker protein showed that CFTR, CC26 and AQP4 are not expressed in olfactory neurones. CFTR is expressed in sustentacular cells of the VNO and olfactory epithelium as well as blood vessels of the underlying mucosa, and VNO (but not Bowman's) glands; a similar pattern (excluding blood vessels) is present for AQP2; AQP4 is found in the two chemosensory epithelia and in Bowman's glands. AQP3 is expressed in the olfactory epithelium and the associated Bowman's glands, but not in the VNO chemosensory epithelium and glands. AQP5 is expressed in the olfactory epithelium and both Bowman's and VNO glands. These results indicate that water/ions handling as well as antioxidant mechanisms operating at the surface and/or inside the nose chemosensory epithelia start developing in utero and are maintained up to sexual maturity. © 2011 Wiley Periodicals, Inc. Develop Neurobiol, 2011

    The diffuse chemosensory system: exploring the iceberg toward the definition of functional roles.

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    The diffuse chemosensory system (DCS) is an anatomical structure composed of solitary chemosensory cells (SCCs, also called solitary chemoreceptor cells), which have analogies with taste cells but are not aggregated in buds. The concept of DCS has been advanced, after the discovery that cells similar to gustatory elements are present in several organs. The elements forming the DCS share common morphological and biochemical characteristics with the taste cells located in taste buds of the oro-pharyngeal cavity but they are localized in internal organs. In particular, they may express molecules of the chemoreceptorial cascade (e.g. trans-membrane taste receptors, the G-protein alpha-gustducin, PLCbeta2, TRPM5). This article will focus on the mammalian DCS in apparatuses of endodermic origin (i.e. digestive and respiratory systems), which is composed of an enormous number of sensory elements and presents a multiplicity of morphological aspects. Recent research has provided an adequate description of these elements, but the functional role for the DCS in these apparatuses is unknown. The initial findings led to the definition of a DCS structured like an iceberg, with a mysterious "submerged" portion localized in the distal part of endodermic apparatuses. Recent work has focussed on the discovery of this submerged portion, which now appears less puzzling. However, the functional roles of the different cytotypes belonging to the DCS are not well known. Recent studies linked chemosensation of the intraluminal content to local control of absorptive and secretory (exocrine and endocrine) processes. Control of the microbial population and detection of irritants seem to be other possible functions of the DCS. In the light of these new findings, the DCS might be thought to be involved in a wide range of diseases of both the respiratory (e.g. asthma, chronic obstructive pulmonary disease, cystic fibrosis) and digestive apparatuses (absorptive or secretive diseases, dysmicrobism), as well as in systemic diseases (e.g. obesity, diabetes). A description of the functional roles of the DCS might be a first step toward the discovery of therapeutic approaches which target chemosensory mechanisms

    The ultrastructure of nasal mucosa in children with asthma.

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    Nasal brushing has sometimes been used to characterize some affections of the respiratory tract, but seldom employed in chronic diseases such as asthma, for the possible presence of cellular inflammation in the small specimens used for electron microscopy. The present study evaluated the ultrastructure of epithelial cells obtained by nasal brushing in 11 allergic children with asthma, before and after staying in an environment free of allergens, usually implicated in the genesis of inflammatory events. The ultrastructural alterations of the nasal mucosa have been graded on the basis of their severity. Grade I lesions were characterized by well-differentiated mucous and ciliated cells. The ciliated cells appeared usually well preserved but decreased in number. In grade II lesions, most of the epithelial surface was covered by mucous cells. A further phenotype composed of poorly differentiated ciliated or mucous cells was detected. Grade III lesions showed aspects of depletion of the ciliated and mucous cells. The epithelium was largely composed of undifferentiated cells. Furthermore, the comparison of specimens at 2 different times of sampling did not differ. The data demonstrate that in allergic children with asthma, the nasal mucosa showed ultrastructural changes, which appeared to be unmodifiable during a prolonged stay in an environment free of allergens. Moreover, the nasal epithelium may provide a convenient sampling site, allowing grade of mucosal damage, with the benefit that the brush method is minimally invasive and avoids complications related to bronchoscopic examination
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