ATP-stimulated electrolyte and mucin secretion in the human intestinal goblet cell line HT29-C1.16E

1994 ◽  
Vol 137 (2) ◽  
Author(s):  
D. Merlin ◽  
C. Augeron ◽  
X.-Y. Tien ◽  
X. Guo ◽  
C.L. Laboisse ◽  
...  
Keyword(s):  
Cell Line ◽  
Goblet Cell ◽  
Mucin Secretion

Purinergic and cholinergic agonists induce exocytosis from the same granule pool in HT29-Cl.16E monolayers

1999 ◽  
Vol 276 (4) ◽  
pp. C907-C914 ◽  
Author(s):  
C. A. Bertrand ◽  
C. L. Laboisse ◽  
U. Hopfer
Keyword(s):  
Cell Line ◽  
Goblet Cell ◽  
Recovery Period ◽  
Impedance Method ◽  
Mucin Secretion ◽  
Cholinergic Agonists ◽  
Cholinergic Agonist ◽  
Epithelial Monolayers

Several secretagogues induce mucin secretion in epithelial monolayers, as determined by measuring released granule contents. To assess whether different agonists act on the same granule pool, capacitance changes in intact monolayers of the goblet cell line HT29-Cl.16E were measured by a novel impedance method. Apical ATP (purinergic agonist) and basolateral carbachol (cholinergic agonist) induce rapid exocytosis with maximal capacitance changes within 3 min. The maximal levels of exocytosis that can be induced by optimal concentrations of either agonist are the same and produce a 30–40% increase in total monolayer capacitance. When ATP and carbachol are applied simultaneously, the magnitude of exocytosis is unchanged from the single-secretagogue level. The recovery of capacitance to baseline (endocytosis) is significantly faster after ATP stimulation than after carbachol stimulation. When ATP and carbachol are applied sequentially at doses that give maximal exocytosis, the magnitude of the capacitance increase produced by the second secretagogue is less than or equal to that of the capacitance decrease during the recovery period. Together, these data suggest that purinergic and cholinergic agonists act on the same granule pool.

Bradykinin modulates mucin secretion but not synthesis from an intestinal goblet cell line

1994 ◽  
Vol 42 (3-4) ◽  
pp. 141-145 ◽  
Author(s):  
C. Michael Stanley ◽  
Thomas E. Phillips
Keyword(s):  
Cell Line ◽  
Goblet Cell ◽  
Mucin Secretion

scholarly journals P2u purinoceptor regulation of mucin secretion in SPOC1 cells, a goblet cell line from the airways

1996 ◽  
Vol 316 (3) ◽  
pp. 943-951 ◽  
Author(s):  
Lubna H. ABDULLAH ◽  
Shannon W. DAVIS ◽  
Lauranell BURCH ◽  
Mitsuo YAMAUCHI ◽  
Scott H. RANDELL ◽  
...  
Keyword(s):  
Molecular Mass ◽  
Cell Line ◽  
Goblet Cell ◽  
Gradient Centrifugation ◽  
Buoyant Density ◽  
Dose Effect ◽  
Mucin Secretion ◽  
Major Band ◽  
Regulatory Pathways ◽  
Cscl Density Gradient

The SPOC1 cell, a novel goblet cell line derived from rat trachea, was tested for its ability to exhibit regulated mucin secretion in response to purinergic (P2) agonists. High-molecular-mass glycoconjugates (HMMGs) purified by CsCl-density-gradient centrifugation had a buoyant density of 1.45 g/ml. The purified HMMG material exhibited a single major band with an apparent molecular mass of greater than 1000 kDa in SDS/ polyacrylamide gels stained with silver or blotted and stained with soya-bean agglutinin. [3H]HMMG was resistant to proteoglycan-degrading enzymes, but was susceptible to neuraminidase. The HMMG was approx. 91% carbohydrate by weight, and the glycosides were O-linked. The HMMG amino acid composition was enriched in Ser and Thr (sum 27%). Thus SPOC1-cell HMMG possess the characteristics of mucin. Mucin secretion by SPOC1 cells, grown on permeable supports and perfused luminally, was stimulated by ATP, UTP and adenosine 5′-[γ-thio]triphosphate (100 μM) 4–5-fold over a baseline of 4 ng/min. The three dose–effect relations were nearly identical (K0.5 ~4 μM). SPOC1 cells grown on plastic and rat tracheal epithelial primary cells responded similarly to ATP and/or UTP. SPOC1 cells failed to respond to other purinergic agonists, either luminally or serosally, and consequently seem to possess an apical membrane P2u purinoceptor. SPOC1-cell total RNA was probed for P2u purinoceptor mRNA. Using conserved primers for both reverse transcriptase and PCR, a single band of the predicted size was observed, which had a nucleotide base sequence identical with the rat P2u purinoceptor mRNA. Thus SPOC1 cells secrete mucin under the control of a P2u purinoceptor; they should prove useful in dissecting the associated cellular regulatory pathways.

scholarly journals Subchronic exposure to acrylamide affects colon mucin secretion in juvenile wistar rats

2016 ◽  
Vol 68 (3) ◽  
pp. 641-649 ◽  
Author(s):  
Ivana Koledin ◽  
Renata Kovac ◽  
Vesna Rajkovic ◽  
Milica Matavulj
Keyword(s):  
Adverse Effect ◽  
Goblet Cell ◽  
Wistar Rats ◽  
Goblet Cells ◽  
Mucin Secretion ◽  
Human Diet ◽  
Mucin Production ◽  
High Carbohydrate ◽  
Antibody Staining ◽  
Male Wistar Rats

Acrylamide (AA) is an important industrial chemical worldwide. AA also forms naturally in many high-carbohydrate foods (bread, French fries, coffee, etc.) when they are heated. Since AA is ubiquitous in the human diet, and more than one-third of the calories we take in each day come from foods with detectable levels of acrylamide, the aim of this study was to determine the effect of subchronic AA treatment on colon goblet cell mucin secretion. Male Wistar rats were gavaged with AA for 5 days a week for 21 days. The animals were divided into three groups that were gavaged with different AA concentrations (0, 25, 50 mg/kg/day). Colon samples were processed for histochemical (PAS-AB, HID-AB) and immunohistochemical (anti-rat MUC2 antibody) staining to visualize mucins in the goblet cells. AA treatment showed an alteration in mucin production and secretion in that the amount of all investigated mucin types dropped. More prominent changes were detected in the upper crypt part where a decreased number of goblet cell was observed. AA treatment elicited a significant reduction in neutral mucins, while acidic mucins showed linearly decreasing trend with respect to AA doses. Also, a linear reduction of MUC2 mucins was noticed. Sulfomucins were absent in the colon lower crypt in all experimental groups, while in the upper crypt both sulfo- and sialomucins were significantly decreased. The results of our study point to changes in the synthesis, differentiation and distribution of mucins after AA treatment, which can have adverse effect on colorectal health.

scholarly journals Recombinant E1-Deleted Adenoviral Vectors Induce Apoptosis in a Rat Airway Epithelial Mucous Goblet Cell Line

1999 ◽  
Vol 81 (1) ◽  
pp. 73-78
Author(s):  
Shinji Teramoto ◽  
Takeshi Matsuse ◽  
Hirotoshi Matsui ◽  
Eijiro Ohga ◽  
Takeo Ishii ◽  
...  
Keyword(s):  
Cell Line ◽  
Goblet Cell ◽  
Adenoviral Vectors ◽  
Airway Epithelial

Protein kinase C-epsilon is the likely mediator of mucin exocytosis in human colonic cell lines

1997 ◽  
Vol 272 (1) ◽  
pp. G31-G37 ◽  
Author(s):  
D. H. Hong ◽  
J. F. Forstner ◽  
G. G. Forstner
Keyword(s):  
Protein Kinase C ◽  
Cell Line ◽  
Cell Lines ◽  
Mucin Secretion ◽  
T84 Cells ◽  
Kinase C ◽  
Pkc Epsilon ◽  
Pkc Alpha ◽  
Colonic Cells ◽  
Ht 29

The phorbol ester, phorbol 12-myristate 13-acetate (PMA), induces mucin secretion in the colonic tumor cell line T84 in a Ca(2+)-independent manner. To determine whether a specific protein kinase C (PKC) isoform is involved in colonic cells, we compared PMA-dependent mucin secretion by three human colonic tumor cell lines (T84, HT-29/A1, and LS 180) with the expression of PKC isoforms alpha, beta, delta, epsilon, and zeta, previously identified in human colon (L. A. Davidson, Y. H. Jiang, J. D. Derr, H. Aukema, J. R. Lupton, and R. S. Chapkin. Arch. Biochem. Biophys. 312:547-553, 1994). In each cell line PMA (10(-7) M) caused mucin secretion within 30 min. PMA-dependent mucin secretion was three to four times greater from HT-29/A1 and T84 cells than from LS 180 cells. All three-cell lines contained mRNA for PKC-alpha, PKC-epsilon, and PKC-zeta but not PKC-beta or -delta. Each cell line also expressed PKC-alpha, -epsilon, and -zeta protein. PKC-epsilon expression (mRNA and protein) was three to four times greater in HT-29/A1 and T84 cells than in LS 180 cells, correlating with PMA-responsive mucin secretion, whereas all cell lines contained similar levels of PKC-alpha mRNA and protein. When cells were stimulated by PMA, only PKC-epsilon was translocated from cytosol to membrane fractions early enough to stimulate mucin secretion. Because PKC-epsilon is also a Ca(2+)-independent isoform, it is likely to mediate mucin exocytosis in colonic cells.

scholarly journals Synthesis and secretion of mucin by the human colonic tumour cell line LS180

1994 ◽  
Vol 302 (1) ◽  
pp. 111-118 ◽  
Author(s):  
D J McCool ◽  
J F Forstner ◽  
G G Forstner
Keyword(s):  
Cell Line ◽  
Carcinoma Cell ◽  
Tumour Cell Line ◽  
Mucin Secretion ◽  
Vicia Villosa ◽  
Golgi Transport ◽  
Sds Page ◽  
Colonic Tumour ◽  
Isolectin B4 ◽  
Time Point

Pulse-chase labelling experiments were performed using the mucin-producing colonic carcinoma cell line LS180. Cells were pulsed with [3H]threonine or [3H]glucosamine and chased in complete media without radiolabel for various lengths of time. From cell and media extracts obtained at each time point, mucin proteins were immunoprecipitated with specific anti-mucin antibodies and analysed by SDS/PAGE and fluorography. At short labelling times with [3H]threonine, without chase, a monomeric thiol-reduction-resistant mucin precursor of apparent molecular mass > 670 kDa was identified. The precursor, in contrast to oligomeric species, was not labelled by [3H]glucosamine but exhibited binding to Vicia villosa isolectin B4, suggesting the presence of some core GalNAc residues. Treatment with tunicamycin to inhibit N-glycosylation had no effect on the apparent mass of the precursor. Identity of the mucin antigen with MUC2 mucin was established by immunoprecipitation with antibodies specific for a MUC2 tandem repeat and C-terminal regions. With increasing chase time the precursor was replaced by thiol-reduction-sensitive mucin oligomers that reached peak intracellular radiolabelling with [3H]threonine by 2 h of chase, and then declined. Only oligomeric mucin was secreted into the medium. Secretion of [3H]threonine-labelled mucin was detectable after 2 h of chase and increased as the cytoplasmic mucin label declined. Monensin inhibited [3H]glucosamine incorporation, sialylation and baseline (non-regulated) mucin secretion without affecting initial [3H]threonine incorporation or oligomerization. Oligomerization and Golgi transport are therefore essential early steps in MUC2 mucin secretion. Oligomerization may follow some core O-glycosylation with GalNAc, but precedes elongation of oligosaccharide chains.

Calcium signaling in human airway goblet cells following purinergic activation

2007 ◽  
Vol 292 (1) ◽  
pp. L92-L98 ◽  
Author(s):  
Andrea H. Rossi ◽  
Wendy C. Salmon ◽  
Michael Chua ◽  
C. William Davis
Keyword(s):  
Goblet Cell ◽  
Primary Cultures ◽  
Goblet Cells ◽  
Pulmonary Diseases ◽  
Mucin Secretion ◽  
Peak Response ◽  
General Importance ◽  
Intracellular Ca ◽  
High Doses ◽  
Purinergic Stimulation

Despite the general importance of Ca2+ signaling in signal transduction, and of goblet cell mucin hypersecretion in inflammatory pulmonary diseases, measurement of airway goblet cell intracellular Ca2+ (Ca[Formula: see text]) has not been reported. In this article, we describe the results of experiments measuring Ca[Formula: see text] in primary cultures of human bronchial goblet cells after stimulation with the purinergic agonist adenosine 5′- O-(3-thiotriphosphate) (ATPγS) and phorbol 12-myristate 13-acetate (PMA). Ca2+ signaling in human goblet cells after purinergic stimulation follows the classic paradigm of a Ca[Formula: see text] transient from a basal activity of 110 nM to a peak response of 260.1 ± 41.2 nM within 2 min, followed by a long superbasal plateau (155.3 ± 0.2 nM) between 10 and 15 min. The rise in Ca[Formula: see text] appears to result from a mobilization of intracellular stores, because the transient was nearly abolished by inhibition of PLC with the phosphatidylinositol-specific PLC inhibitor U-73122, and it was not affected significantly by removal of extracellular Ca2+. Loading goblet cells with BAPTA inhibited the ATPγS-induced Ca2+ transient by 86.0 ± 13.1%, relative to control. Finally, in contrast to the massive effects of high doses of PMA (300 nM) on mucin secretion from goblet cells, phorbol ester stimulated a small (27.1 ± 7% of the ATPγS control peak), brief rise in Ca[Formula: see text]. This diminutive signal likely denotes a local Ca2+ gradient, which may be associated with the mucin granule exocytotic process.

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