The Experts below are selected from a list of 312 Experts worldwide ranked by ideXlab platform
Jeffrey J Wine - One of the best experts on this subject based on the ideXlab platform.
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the normal trachea is cleaned by muc5b mucin bundles from the Submucosal Glands coated with the muc5ac mucin
Biochemical and Biophysical Research Communications, 2017Co-Authors: Anna Ermund, Lauren N Meiss, Ana M Rodriguezpineiro, Andrea Bahr, Harriet Nilsson, Sergio Trillomuyo, Caroline Ridley, David J Thornton, Jeffrey J WineAbstract:To understand the mucociliary clearance system, mucins were visualized by light, confocal and electron microscopy, and mucus was stained by Alcian blue and tracked by video microscopy on tracheal explants of newborn piglets. We observed long linear mucus bundles that appeared at the Submucosal gland openings and were transported cephalically. The mucus bundles were shown by mass spectrometry and immunostaining to have a core made of MUC5B mucin and were coated with MUC5AC mucin produced by surface goblet cells. The transport speed of the bundles was slower than the airway surface liquid flow. We suggest that the goblet cell MUC5AC mucin anchors the mucus bundles and thus controls their transport. Normal clearance of the respiratory tree of pigs and humans, both rich in Submucosal Glands, is performed by thick and long mucus bundles.
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secretion rates of human nasal Submucosal Glands from patients with chronic rhinosinusitis or cystic fibrosis
American Journal of Rhinology & Allergy, 2015Co-Authors: Peter H Hwang, Jin Hyeok Jeong, Jeffrey J WineAbstract:BACKGROUND: A majority of patients with cystic fibrosis (CF) have chronic rhinosinusitis (CRS) and/or nasal polyps, both of which may be secondary to reduced fluid secretion from nasal Submucosal Glands. OBJECTIVE: To determine whether decreased fluid secretion from nasal Submucosal Glands also occurs in patients without CF and with CRS. METHODS: Inferior turbinates of the nasal cavity were harvested from controls, subjects with CRS, and subjects with CF (n = 5-7 per group). The secretion rates of the nasal Submucosal Glands of the three groups in response to carbachol and forskolin were measured by using time lapse digital imaging of mucus bubbles from single Glands as they formed on the mucosal surface under oil. RESULTS: Carbachol-stimulated secretion rates were the following: controls, 1670 ± 381 pl·min(-1)·gland(-1); CRS, 965 ± 440 pl·min(-1)·gland(-1); and CF, 933 ± 588 pl·min(-1)·gland(-1) (p = 0.23, Kruskal-Wallis test). Forskolin-stimulated secretion rates were the following: controls, 229 ± 14 pl·min(-1)·gland(-1); CRS, 154 ± 48 pl·min(-1)·gland(-1); and CF, 22 ± 15 pl·min(-1)·gland(-1) (p = 0.008, Kruskal-Wallis test). The ratio of the average secretion rate induced by forskolin to that induced by carbachol was 13.7% in the controls, and 15.9% in CRS and 2.3% in CF groups. CONCLUSION: The only significant difference in this small study was decreased forskolin-stimulated secretion in subjects with CF relative to the other subjects. However, there was a trend toward reduced carbachol-stimulated secretion rates in subjects with CRS and with and without CF relative to controls. Additional studies are needed to determine if nasal Submucosal gland hyposecretion occurs in CRS either as a contributor to or as a consequence of CRS pathogenesis.
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Mucociliary clearance and Submucosal gland secretion in the ex vivo ferret trachea
American Journal of Physiology-lung Cellular and Molecular Physiology, 2014Co-Authors: Jin Hyeok Jeong, Peter H Hwang, Jeffrey J WineAbstract:In many species Submucosal Glands are an important source of tracheal mucus, but the extent to which mucociliary clearance (MCC) depends on gland secretion is unknown. To explore this relationship,...
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properties of substance p stimulated mucus secretion from porcine tracheal Submucosal Glands
American Journal of Physiology-lung Cellular and Molecular Physiology, 2011Co-Authors: Monal Khansaheb, Jae Young Choi, Yumi Yang, Mauri E Krouse, Jeffrey J WineAbstract:Human and pig airway Submucosal Glands secrete mucus in response to substance P (SubP), but in pig tracheal Glands the response to SubP is >10-fold greater than in humans and shares features with c...
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measurement of fluid secretion from intact airway Submucosal Glands
Methods of Molecular Biology, 2011Co-Authors: Jeffrey J Wine, Monal Khansaheb, Jae Young Choi, Mauri E Krouse, Juan P Ianowski, Jin V Wu, Toshiya Irokawa, John W Hanrahan, A W CuthbertAbstract:: Human airways are kept sterile by a mucosal innate defense system that includes mucus secretion. Mucus is secreted in healthy upper airways primarily by Submucosal Glands and consists of defense molecules mixed with mucins, electrolytes, and water and is also a major component of sputum. Mucus traps pathogens and mechanically removes them via mucociliary clearance while inhibiting their growth via molecular (e.g., lysozyme) and cellular (e.g., neutrophils, macrophages) defenses. Fluid secretion rates of single Glands in response to various mediators can be measured by trapping the primary gland mucus secretions in an oil layer, where they form spherical bubbles that can be optically measured at any desired interval to provide detailed temporal analysis of secretion rates. The composition and properties of the mucus (e.g., solids, viscosity, pH) can also be determined. These methods have now been applied to mice, ferrets, cats, pigs, sheep, and humans, with a main goal of comparing gland secretion in control and CFTR-deficient humans and animals.
Stephen T Ballard - One of the best experts on this subject based on the ideXlab platform.
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fluid secretion by Submucosal Glands of the tracheobronchial airways
Respiratory Physiology & Neurobiology, 2007Co-Authors: Stephen T Ballard, Domenico SpadaforaAbstract:Submucosal Glands of the tracheobronchial airways provide the important functions of secreting mucins, antimicrobial substances, and fluid. This review focuses on the ionic mechanism and regulation of gland fluid secretion and examines the possible role of gland dysfunction in the lethal disease cystic fibrosis (CF). The fluid component of gland secretion is driven by the active transepithelial secretion of both Cl− and HCO3− by serous cells. Gland fluid secretion is neurally regulated with acetylcholine, substance P, and vasoactive intestinal peptide (VIP) playing prominent roles. The cystic fibrosis transmembrane conductance regulator (CFTR) is present in the apical membrane of gland serous cells and mediates the VIP-induced component of liquid secretion whereas the muscarinic component of liquid secretion appears to be at least partially CFTR-independent. Loss of CFTR function, which occurs in CF disease, reduces the capacity of Glands to secrete fluid but not mucins. The possible links between the loss of fluid secretion capability and the complex airway pathology of CF are discussed.
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liquid secretion properties of airway Submucosal Glands
The Journal of Physiology, 2004Co-Authors: Stephen T Ballard, S K InglisAbstract:The tracheobronchial Submucosal Glands secrete liquid that is important for hydrating airway surfaces, supporting mucociliary transport, and serving as a fluid matrix for numerous secreted macromolecules including the gel-forming mucins. This review details the essential structural elements of airway Glands and summarizes what is currently known regarding the ion transport processes responsible for producing the liquid component of gland secretion. Liquid secretion most likely arises from serous cells and is principally under neural control with muscarinic agonists, substance P, and vasoactive intestinal peptide (VIP) functioning as effective secretogogues. Liquid secretion is driven by the active transepithelial secretion of both Cl− and HCO3− and at least a portion of this process is mediated by the cystic fibrosis transmembrane conductance regulator (CFTR), which is highly expressed in Glands. The potential role of Submucosal Glands in cystic fibrosis lung disease is discussed.
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CFTR involvement in chloride, bicarbonate, and liquid secretion by airway Submucosal Glands.
American Journal of Physiology-lung Cellular and Molecular Physiology, 1999Co-Authors: Stephen T Ballard, Laura Trout, Zsuzsa Bebok, Eric J. Sorscher, Angela D. CrewsAbstract:Previous studies demonstrated that ACh-induced liquid secretion by porcine bronchi is driven by active Cl− and H CO 3 − secretion. The present study was undertaken to determine whether this process was localized to Submucosal Glands and mediated by the cystic fibrosis transmembrane conductance regulator (CFTR). When excised, cannulated, and treated with ACh, porcine bronchi secreted 15.6 ± 0.6 μl ⋅ cm−2 ⋅ h−1. Removal of the surface epithelium did not significantly affect the rate of secretion, indicating that the source of the liquid was the Submucosal Glands. Pretreatment with diphenylamine-2-carboxylate, a relatively nonselective Cl−-channel blocker, significantly reduced liquid secretion by 86%, whereas pretreatment with DIDS, which inhibits a variety of Cl− channels but not CFTR, had no effect. When bronchi were pretreated with glibenclamide or 5-nitro-2-(3-phenylpropylamino)benzoic acid (both inhibitors of CFTR), the rate of ACh-induced liquid secretion was significantly reduced by 39 and 91%, respe...
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in situ visualization of bronchial Submucosal Glands and their secretory response to acetylcholine
American Journal of Physiology-lung Cellular and Molecular Physiology, 1997Co-Authors: S K Inglis, M R Corboz, A E Taylor, Stephen T BallardAbstract:Airway Submucosal Glands secrete both macromolecules and liquid, yet the mechanisms by which these substances are secreted are not well understood. In this study, a video microscope was used to directly visualize the Submucosal Glands in isolated porcine distal bronchi and to observe their responses to acetylcholine (ACh), a glandular secretagogue. Submucosal Glands were classified as either "antral," "linear," or "convoluted" Glands based on the morphology of their terminal collecting ducts. Because antral duct Glands were most easily visualized, the response to ACh was studied in detail in this gland type. Within 5-10 s after addition of 10 microM ACh, the cross-sectional area of the gland duct openings to the airway surface increased severalfold but returned to pre-ACh dimensions within 1 min. Between 30 s and 10 min after ACh addition, spherical particles (1-10 microm) entered the antral ducts from distal acini and exited through the duct openings to the airway surface. Some of the particles were reta...
John F Engelhardt - One of the best experts on this subject based on the ideXlab platform.
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lysozyme secretion by Submucosal Glands protects the airway from bacterial infection
American Journal of Respiratory Cell and Molecular Biology, 2005Co-Authors: Rana Dajani, Yulong Zhang, Peter J Taft, Sue M Travis, Timothy D Starner, Ansgar Olsen, Joseph Zabner, Michael J Welsh, John F EngelhardtAbstract:Submucosal Glands are abundant (∼ 1 gland/mm2) secretory structures in the tracheobronchial airways of the human lung. Because Submucosal Glands express antibacterial proteins, it has been proposed that they contribute to lung defense. However, this concept is challenged by the fact that mice do not have Submucosal Glands in their bronchial airways, yet are quite resistant to bacterial lung infection. The contribution of airway Submucosal Glands to host defense is also debated as a pathophysiologic component of cystic fibrosis lung disease. Here, we asked whether Submucosal Glands protect airways against bacterial infection. By comparing tracheal xenograft airways with and without Glands, we found that the presence of Glands enhanced bacterial killing in vivo and by airway secretions in vitro. Moreover, immunodepletion studies suggested that lysozyme is a major antibacterial component secreted by Submucosal Glands. These studies provide evidence that Submucosal Glands are a major source of antibacterials ...
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muc5b and muc7 are differentially expressed in mucous and serous cells of Submucosal Glands in human bronchial airways
American Journal of Respiratory Cell and Molecular Biology, 1998Co-Authors: Prerna Sharma, Lorita Dudus, Peter Aadal Nielsen, Henrik Clausen, James R Yankaskas, Michael A Hollingsworth, John F EngelhardtAbstract:Mucins are high molecular-weight glycoproteins involved in the protection and lubrication of respiratory, gastrointestinal, and reproductive tracts. Hypersecretory diseases such as cystic fibrosis (CF), chronic bronchitis, and asthma result in dysregulated levels of mucin production stemming from increased abundance of mucin-secreting cell types in the surface airway epithelium and Submucosal Glands. The isolation of at least nine mucin genes has prompted studies to characterize the cellular expression patterns of these mucins in normal and diseased tissues. In the present study, in situ hybridization and immunocytochemical methods were used to determine the cellular distribution of MUC5B and MUC7 expression in CF and non-CF human bronchus. Our findings indicate that MUC5B and MUC7 have expression patterns in human bronchial airways that are limited exclusively to Submucosal Glands. Specifically, MUC5B expression was confined to all mucous tubules, whereas MUC7 expression was seen in a subset of lysozyme ...
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Developmental expression patterns of CFTR in ferret tracheal surface airway and Submucosal gland epithelia.
American Journal of Respiratory Cell and Molecular Biology, 1996Co-Authors: Anil Sehgal, Asaf Presente, John F EngelhardtAbstract:Submucosal Glands are a major site of cystic fibrosis transmembrane conductance regulator (CFTR) expression in the human airway and may play an important role in the pathogenesis of cystic fibrosis. In humans, strategies for gene targeting to Submucosal Glands will likely be dependent on in utero gene transfer to Submucosal gland progenitors because of the inaccessibility of these regions from the fully developed airway. However, little is known about the ontogeny of CFTR gene expression in developing Submucosal Glands and the potential functional role(s) CFTR may have during gland development. To this end, we describe the partial cloning of the ferret CFTR cDNA which was used for in situ mRNA localization studies in developing ferret trachea. The ferret animal model is attractive for studies pertaining to the development of tracheal Submucosal Glands because postnatal tracheal development in this species mirrors in utero gland development in humans. Sequence analysis of the first nucleotide binding domai...
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adenovirus mediated gene transfer to human bronchial Submucosal Glands using xenografts
American Journal of Physiology-lung Cellular and Molecular Physiology, 1995Co-Authors: J M Pilewski, John F Engelhardt, Joseph E Bavaria, Larry R Kaiser, James M Wilson, Steven M AlbeldaAbstract:The cystic fibrosis (CF) transmembrane conductance regulator has been localized to both Submucosal Glands and surface epithelium, suggesting that both glandular and surface epithelium may be important targets for gene therapy. To determine the distribution and efficiency of recombinant adenovirus-mediated gene transfer to human airway Submucosal Glands, an in vivo model was developed by heterotopically transplanting human bronchial segments from both normal and CF lung tissue into severe combined immunodeficient mice. A serotype 5 E1-deleted recombinant adenovirus containing a lacZ reporter gene driven by the cytomegalovirus promoter (H5.010CMVlacZ) was used to infect the xenografts. Transgene expression was correlated with three factors: 1) viral titer, 2) penetration of microspheres, and 3) dwell time of the viral instillate. At viral titers ranging from 10(8) to 10(11) plaque forming units/ml, expression of the lacZ gene was observed in a subpopulation of epithelial cells within approximately 40% of su...
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adenovirus mediated gene transfer to human brochial Submucosal Glands using xenografts
American Journal of Physiology-lung Cellular and Molecular Physiology, 1995Co-Authors: J M Pilewski, John F Engelhardt, Joseph E Bavaria, Larry R Kaiser, James M Wilson, Steven M AlbeldaAbstract:The cystic fibrosis (CF) transmembrane conductance regulator has been localized to both Submucosal Glands and surface epithelium, suggesting that both glandular and surface epithelium may be important targets for gene therapy. To determine the distribution and efficiency of recombinant adenovirus-mediated gene transfer to human airway Submucosal Glands, an in vivo model was developed by heterotopically transplanting human bronchial segments from both normal and CF lung tissue into severe combined immunodeficient mice. A serotype 5 E1-deleted recombinant adenovirus containing a lacZ reporter gene driven by the cytomegalovirus promoter (H5.010CMVlacZ) was used to infect the xenografts. Transgene expression was correlated with three factors: 1) viral titer, 2) penetration of microspheres, and 3) dwell time of the viral instillate. At viral titers ranging from 10 8 to 10 11 plaque forming units/ml, expression of the lacZ gene was observed in a subpopulation of epithelial cells within ∼ 40% of Submucosal Glands, with more efficient gene transfer to the ducts than the secretory tubules. Within individual Glands, gene transfer varied from < 1% to ∼ 20% of Submucosal cells, including duct, mucous, and serous cells. Adenovirus-sized fluorescent microspheres were found to penetrate only some of the Submucosal Glands, suggesting that the gene transfer efficiency to Submucosal tubules is due to limited viral particle penetration rather than tropism. Gene transfer to surface epithelial cells was inefficient. However, the percentage of transduced surface epithelial cells increased from < 1% to 5-10% as the dwell time of viral solution was increased from 5 min to 1 h, indicating that the time allowed for virus binding and entry is important for gene transfer efficiency. This report presents the first definitive demonstration of adenovirus-mediated transgene delivery into human airway Submucosal Glands
Juan P Ianowski - One of the best experts on this subject based on the ideXlab platform.
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airway Submucosal Glands from cystic fibrosis swine suffer from abnormal ion transport across the serous acini collecting duct and ciliated duct
American Journal of Physiology-lung Cellular and Molecular Physiology, 2020Co-Authors: Xiaojie Luan, Santosh Jagadeeshan, Nikolay Grishchenko, Noman Hassan, Paula Gioino, Alan M Shipley, Terry E Machen, Juan P IanowskiAbstract:: The human airway is protected by an efficient innate defence mechanism that requires healthy secretion of airway surface liquid (ASL) to clear pathogens from the lungs. Most of the ASL in the upper airway is secreted by Submucosal Glands. In cystic fibrosis (CF), the function of airway Submucosal Glands is abnormal, and these abnormalities are attributed to anomalies in ion transport across the epithelia lining the different sections of the Glands that function coordinately to produce the ASL. However, the ion transport properties of most of the anatomical regions of the gland have never been measured, and there is controversy regarding which segments express CFTR. This makes it difficult to determine the glandular abnormalities that may contribute to CF lung disease. Using a non-invasive, extracellular self-referencing ion selective electrode technique, we characterized ion transport properties in all four segments of Submucosal Glands from wild-type and CFTR-/- swine. In wild-type airways the serous acini, mucus tubules, and collecting ducts secrete Cl- and Na+ into the lumen in response to carbachol and forskolin stimulation. The ciliated duct also transports Cl- and Na+ but in the opposite direction, i.e. reabsorption from the ASL, which may contribute to lowering Na+ and Cl- activities in the secreted fluid. In CFTR-/- airways the serous acini, collecting ducts, and ciliated ducts fail to transport ions after forskolin stimulation, resulting in the production of smaller volumes of ASL with normal Cl-, Na+ and K+ concentration.
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interleukin 8 stimulates cftr mediated mucus secretion by Submucosal Glands in swine 869 10
The FASEB Journal, 2014Co-Authors: Haylee Hansvall, Juan P IanowskiAbstract:Cystic Fibrosis (CF) is an autosomal recessive condition caused by mutations in the gene encoding the Cystic Fibrosis Transmembrane conductance Regulator (CFTR) anion channel. As a result epithelial fluid and ion transport is compromised. Many organs are affected in CF patients, but most of the mortality and morbidity arise from airway disease characterized by chronic infection and inflammation. However, the events leading from CFTR mutation to airway disease are not fully understood. Recent data suggest that abnormal bacteria clearance due to abnormal mucus secretion is the pathogenic event that triggers airway disease in CF. Thus we studied the response of airway epithelia to proinflammatory cytokines, such as Interleukin-8 (IL-8), that are released in response to bacterial infection and are elevated in CF airways. Secretion assays and electrophysiological techniques were used to measure the response of Submucosal Glands, the principal mucus source, to IL-8. The mucus secretion rate increased from 0.052...
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measurement of fluid secretion from intact airway Submucosal Glands
Methods of Molecular Biology, 2011Co-Authors: Jeffrey J Wine, Monal Khansaheb, Jae Young Choi, Mauri E Krouse, Juan P Ianowski, Jin V Wu, Toshiya Irokawa, John W Hanrahan, A W CuthbertAbstract:: Human airways are kept sterile by a mucosal innate defense system that includes mucus secretion. Mucus is secreted in healthy upper airways primarily by Submucosal Glands and consists of defense molecules mixed with mucins, electrolytes, and water and is also a major component of sputum. Mucus traps pathogens and mechanically removes them via mucociliary clearance while inhibiting their growth via molecular (e.g., lysozyme) and cellular (e.g., neutrophils, macrophages) defenses. Fluid secretion rates of single Glands in response to various mediators can be measured by trapping the primary gland mucus secretions in an oil layer, where they form spherical bubbles that can be optically measured at any desired interval to provide detailed temporal analysis of secretion rates. The composition and properties of the mucus (e.g., solids, viscosity, pH) can also be determined. These methods have now been applied to mice, ferrets, cats, pigs, sheep, and humans, with a main goal of comparing gland secretion in control and CFTR-deficient humans and animals.
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substance p stimulates cftr dependent fluid secretion by mouse tracheal Submucosal Glands
Pflügers Archiv: European Journal of Physiology, 2008Co-Authors: Jae Young Choi, Jeffrey J Wine, Juan P Ianowski, John W HanrahanAbstract:The mucosa of the proximal airways defends itself and the lower airways from inhaled irritants such as capsaicinoids, allergens, and infections by several mechanisms. Sensory nerves monitor the luminal microenvironment and release the tachykinin substance P (SP) to stimulate mucus secretion. Here, we have studied the role of the cystic fibrosis transmembrane conductance regulator (CFTR) in SP stimulation by comparing mouse airway Submucosal gland responses in wild-type (WT) and CFTR−/− mice. Capsaicinoids (chili pepper oil) increased fluid secretion by Glands from WT mice five-fold, and this response was abolished by exposing the basolateral aspect of the tracheas to L-732,138 (10 μmol/l), a specific antagonist of the neurokinin-1 receptor. Secretion was also stimulated 25-fold by basolateral application of SP, and this response was strongly inhibited by the CFTR inhibitor CFTRinh172. In contrast, Submucosal Glands from CFTR knockout mice failed to secrete when stimulated by SP (1 μmol/l), although those from wild-type control littermates were responsive. SP stimulation of wild-type Glands was also abolished by clotrimazole (25 μmol/l), a blocker of Ca2+-activated K+ channels. These results indicate that SP mediates local responses to capsaicinoids through a mechanism involving coordinated activation of CFTR and K+ channels. To our knowledge, this is the first study in which CFTR-dependent responses to substance P have been directly demonstrated. Since CFTR regulation is qualitatively similar in human and mouse Glands, loss of this local regulation in CF may contribute to reduced innate defenses in CF airways.
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mucus secretion by single tracheal Submucosal Glands from normal and cystic fibrosis transmembrane conductance regulator knockout mice
The Journal of Physiology, 2007Co-Authors: Juan P Ianowski, Jae Young Choi, Jeffrey J Wine, John W HanrahanAbstract:Submucosal Glands line the cartilaginous airways and produce most of the antimicrobial mucus that keeps the airways sterile. The Glands are defective in cystic fibrosis (CF), but how this impacts airway health remains uncertain. Although most CF mouse strains exhibit mild airway defects, those with the C57Bl/6 genetic background have increased airway pathology and susceptibility to Pseudomonas. Thus, they offer the possibility of studying whether, and if so how, abnormal Submucosal gland function contributes to CF airway disease. We used optical methods to study fluid secretion by individual Glands in tracheas from normal, wild-type (WT) mice and from cystic fibrosis transmembrane conductance regulator (CFTR) knockout mice (Cftrm1UNC/Cftrm1UNC; CF mice). Glands from WT mice qualitatively resembled those in humans by responding to carbachol and vasoactive intestinal peptide (VIP), although the relative rates of VIP- and forskolin-stimulated secretion were much lower in mice than in large mammals. The pharmacology of mouse gland secretion was also similar to that in humans; adding bumetanide or replacement of HCO3− by Hepes reduced the carbachol response by ∼50%, and this inhibition increased to 80% when both manoeuvres were performed simultaneously. It is important to note that Glands from CFTR knockout mice responded to carbachol but did not secrete when exposed to VIP or forskolin, as has been shown previously for Glands from CF patients. Tracheal Glands from WT and CF mice both had robust secretory responses to electrical field stimulation that were blocked by tetrodotoxin. It is interesting that local irritation of the mucosa using chili pepper oil elicited secretion from WT Glands but did not stimulate Glands from CF mice. These results clarify the mechanisms of murine Submucosal gland secretion and reveal a novel defect in local regulation of Glands lacking CFTR which may also compromise airway defence in CF patients.
Jae Young Choi - One of the best experts on this subject based on the ideXlab platform.
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vitamin a deficiency induces fluid hyposecretion from the airway Submucosal Glands of mice
Journal of Nutrition, 2012Co-Authors: Jooheon Yoon, Jae Young ChoiAbstract:: Vitamin A deficiency (VAD) alters the phenotype of airway epithelium and attenuates the epithelial defense system, and many studies have reported the association of VAD with respiratory disease. In this study, we investigated changes in Submucosal Glands (SMG) in a mouse model of VAD. C57BL/6 mice were fed a vitamin A-devoid diet and the others were fed a control diet (1.2 mg retinol/kg). The areas of serous and mucous cells of SMG were measured in 4-, 8-, and 20-wk-old male mice. The volume and lysozyme concentration of glandular secretions were also measured. The 2 groups did not differ in body weight or general morbidity at 3-10 wk of age, although serum retinol concentrations were greater in the control mice than in the VAD mice after 4 wk. Upon histological evaluation, we found that the areal ratio of serous cells:total SMG cells was significantly lower after 8 wk in the VAD mice compared with the control mice, although the total area of SMG did not differ between groups throughout the 20-wk experiment. The number of secretory bubbles did not differ between the groups, but total secretion volume was reduced by 35% in 8-wk-old VAD mice compared with controls. Furthermore, the concentration of lysozyme in secretions from 8-wk-old VAD mice was also less than in controls, compounding the effect of diminished secretion volume. In this study, we found serous cell hypotrophy/hypoplasia and dysfunction in VAD mice, which may contribute to the susceptibility to airway infection linked to VAD.
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properties of substance p stimulated mucus secretion from porcine tracheal Submucosal Glands
American Journal of Physiology-lung Cellular and Molecular Physiology, 2011Co-Authors: Monal Khansaheb, Jae Young Choi, Yumi Yang, Mauri E Krouse, Jeffrey J WineAbstract:Human and pig airway Submucosal Glands secrete mucus in response to substance P (SubP), but in pig tracheal Glands the response to SubP is >10-fold greater than in humans and shares features with c...
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measurement of fluid secretion from intact airway Submucosal Glands
Methods of Molecular Biology, 2011Co-Authors: Jeffrey J Wine, Monal Khansaheb, Jae Young Choi, Mauri E Krouse, Juan P Ianowski, Jin V Wu, Toshiya Irokawa, John W Hanrahan, A W CuthbertAbstract:: Human airways are kept sterile by a mucosal innate defense system that includes mucus secretion. Mucus is secreted in healthy upper airways primarily by Submucosal Glands and consists of defense molecules mixed with mucins, electrolytes, and water and is also a major component of sputum. Mucus traps pathogens and mechanically removes them via mucociliary clearance while inhibiting their growth via molecular (e.g., lysozyme) and cellular (e.g., neutrophils, macrophages) defenses. Fluid secretion rates of single Glands in response to various mediators can be measured by trapping the primary gland mucus secretions in an oil layer, where they form spherical bubbles that can be optically measured at any desired interval to provide detailed temporal analysis of secretion rates. The composition and properties of the mucus (e.g., solids, viscosity, pH) can also be determined. These methods have now been applied to mice, ferrets, cats, pigs, sheep, and humans, with a main goal of comparing gland secretion in control and CFTR-deficient humans and animals.
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substance p stimulates cftr dependent fluid secretion by mouse tracheal Submucosal Glands
Pflügers Archiv: European Journal of Physiology, 2008Co-Authors: Jae Young Choi, Jeffrey J Wine, Juan P Ianowski, John W HanrahanAbstract:The mucosa of the proximal airways defends itself and the lower airways from inhaled irritants such as capsaicinoids, allergens, and infections by several mechanisms. Sensory nerves monitor the luminal microenvironment and release the tachykinin substance P (SP) to stimulate mucus secretion. Here, we have studied the role of the cystic fibrosis transmembrane conductance regulator (CFTR) in SP stimulation by comparing mouse airway Submucosal gland responses in wild-type (WT) and CFTR−/− mice. Capsaicinoids (chili pepper oil) increased fluid secretion by Glands from WT mice five-fold, and this response was abolished by exposing the basolateral aspect of the tracheas to L-732,138 (10 μmol/l), a specific antagonist of the neurokinin-1 receptor. Secretion was also stimulated 25-fold by basolateral application of SP, and this response was strongly inhibited by the CFTR inhibitor CFTRinh172. In contrast, Submucosal Glands from CFTR knockout mice failed to secrete when stimulated by SP (1 μmol/l), although those from wild-type control littermates were responsive. SP stimulation of wild-type Glands was also abolished by clotrimazole (25 μmol/l), a blocker of Ca2+-activated K+ channels. These results indicate that SP mediates local responses to capsaicinoids through a mechanism involving coordinated activation of CFTR and K+ channels. To our knowledge, this is the first study in which CFTR-dependent responses to substance P have been directly demonstrated. Since CFTR regulation is qualitatively similar in human and mouse Glands, loss of this local regulation in CF may contribute to reduced innate defenses in CF airways.
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mucus secretion by single tracheal Submucosal Glands from normal and cystic fibrosis transmembrane conductance regulator knockout mice
The Journal of Physiology, 2007Co-Authors: Juan P Ianowski, Jae Young Choi, Jeffrey J Wine, John W HanrahanAbstract:Submucosal Glands line the cartilaginous airways and produce most of the antimicrobial mucus that keeps the airways sterile. The Glands are defective in cystic fibrosis (CF), but how this impacts airway health remains uncertain. Although most CF mouse strains exhibit mild airway defects, those with the C57Bl/6 genetic background have increased airway pathology and susceptibility to Pseudomonas. Thus, they offer the possibility of studying whether, and if so how, abnormal Submucosal gland function contributes to CF airway disease. We used optical methods to study fluid secretion by individual Glands in tracheas from normal, wild-type (WT) mice and from cystic fibrosis transmembrane conductance regulator (CFTR) knockout mice (Cftrm1UNC/Cftrm1UNC; CF mice). Glands from WT mice qualitatively resembled those in humans by responding to carbachol and vasoactive intestinal peptide (VIP), although the relative rates of VIP- and forskolin-stimulated secretion were much lower in mice than in large mammals. The pharmacology of mouse gland secretion was also similar to that in humans; adding bumetanide or replacement of HCO3− by Hepes reduced the carbachol response by ∼50%, and this inhibition increased to 80% when both manoeuvres were performed simultaneously. It is important to note that Glands from CFTR knockout mice responded to carbachol but did not secrete when exposed to VIP or forskolin, as has been shown previously for Glands from CF patients. Tracheal Glands from WT and CF mice both had robust secretory responses to electrical field stimulation that were blocked by tetrodotoxin. It is interesting that local irritation of the mucosa using chili pepper oil elicited secretion from WT Glands but did not stimulate Glands from CF mice. These results clarify the mechanisms of murine Submucosal gland secretion and reveal a novel defect in local regulation of Glands lacking CFTR which may also compromise airway defence in CF patients.
