Ion transport in primary cultures from human sweat gland coils studied with X‐ray microanalysis

Mörk, Ann‐Christin; Hongpaisan, Jarin; Roomans, Godfried M.

doi:10.1006/cbir.1995.1056

Cited by 16 publications

(12 citation statements)

References 12 publications

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“…The present findings show that the Cl and K concentrations in cells from primary cultures from sweat gland ducts were sensitive to stimulation by ATP, but not by UTP. The effect of ATP on the Cl and K concentration has previously been shown in primary cultures from human sweat gland coils (18) and in a sweat gland cell line (30,31). Our findings show that ATP, carbachol, the Ca2+ ionophore A23187, and CAMPhad the same potency for activation of the loss of Cl from primary cultures of duct cells.…”

Section: Discussionsupporting

confidence: 73%

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Regulation of Ion Content in Primary Cultures from Reabsorptive Ducts of Human Sweat Glands Studied by X-ray Microanalysis.

Hongpaisan

Roomans

1998

Cell Struct. Funct.

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Section: Discussionsupporting

confidence: 73%

“…In primary cultures from humansweat gland coils, at 5 min after stimulation, the effect of ATPon the decrease of Cl concentration was approximately 50% of that of carbachol and about the same as that of acetylcholine (18). In NCL-SG3cells, ATP is a very effective regulator of membrane permeability (30).…”

Section: Discussionmentioning

confidence: 95%

Regulation of Ion Content in Primary Cultures from Reabsorptive Ducts of Human Sweat Glands Studied by X-ray Microanalysis.

Hongpaisan

Roomans

1998

Cell Struct. Funct.

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“…The present results confirm that genistein induces the cAMP-dependent activation of DF508 CFTR and suggests further that it is able to increase the Cl -secretion achieved after incubation with 4PBA. It has previously been shown, by X-ray microanalysis, that loss of cellular chlorine (equivalent to Cl -efflux) occurs after stimulation with cAMP (analogues) in, for example, airway epithelial cells [16], tracheal gland cells [17] and sweat gland cells [18] containing the CFTR channel. This loss was absent in airway epithelial cells from CF patients [16], tracheal gland cells treated with antisense CFTR [17] and sweat gland cells and tracheal gland cells in the presence of chloride channel blockers [17±19].…”

Section: Discussionmentioning

confidence: 99%

Activation of DeltaF508 CFTR in a cystic fibrosis respiratory epithelial cell line by 4-phenylbutyrate, genistein and CPX

Andersson¹,

Roomans²

2000

Eur Respir J

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The cellular basis of cystic fibrosis (CF) is a defect in a cyclic adenosine monophosphate (cAMP)‐activated chloride channel (CF transmembrane conductance regulator) in epithelial cells that leads to decreased chloride ion transport and impaired water transport across the cell membrane. This study investigated whether it was possible to activate the defective chloride channel in cystic fibrosis respiratory epithelial cells with 4‐phenylbutyrate (4PBA), genistein and 8‐cyclopentyl‐1,3‐dipropylxanthine (CPX). The CF bronchial epithelial cell line CFBE41o‐, which expresses the ΔF508 mutation, was treated with these agents and loss of Cl‐, indicating Cl‐ efflux, measured by X‐ray microanalysis. 8‐bromo‐cAMP alone did not induce Cl‐ efflux in CFBE41o‐ cells, but after incubation with 4PBA a significant efflux of Cl‐ occurred. Stimulation of cells with a combination of genistein and cAMP also induced Cl‐ efflux, whereas a combination of pretreatment with 4PBA and a combined stimulation with genistein and cAMP induced an even larger Cl‐ efflux. Cl‐ efflux could also be stimulated by CPX, but this effect was not enhanced by 4PBA pretreatment. The ΔF508 mutation leads to impaired processing of the cystic fibrosis transmembrane conductance regulator. The increased efflux of chloride after 4‐phenylbutyrate treatment can be explained by the fact that 4‐phenylbutyrate allows the ΔF508 cystic fibrosis transmembrane conductance regulator to escape degradation and to be transported to the cell surface. Genistein and 8‐cyclopentyl‐1,3‐dipropylxanthine act by stimulating chloride ion efflux by increasing the probability of the cystic fibrosis transmembrane conductance regulator being open. The combination of 4‐phenylbutyrate and genistein may be useful in a potential pharmacological therapy for cystic fibrosis patients with the ΔF508 mutation.

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“…The cellcovered grids were preincubated with one of the Clchannel blockers followed by incubation with acetylcholine or cAMP in the presence of this Cl -channel blocker (table 1). The dose of the agents used in this study is close to the known maximum dose for this or similar cell types [14]. As controls, we used cells incubated with only SRS (for 3 or 5 min), 1% dimethylsulphoxide (DMSO) in SRS, 1% ethanol in SRS, 0.5 mM 9-AC, 50 µM NPPB or 0.2 mM DPC (for 5 or 7 min).…”

Section: Preparation Of Cultured Cells For X-ray Microanalysismentioning

confidence: 99%

Ion transport in cultured pig tracheal submucosal gland acinar cells studied by X-ray microanalysis

Zhang

Roomans²

1997

Eur Respir J

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The submucosal glands of the airway may contribute more to the airway fluid than the surface epithelium. The cellular mechanisms underlying the regulation of electrolyte and water transport in airway submucosal glands are, however, still poorly understood. Therefore, we attempted to establish a cell culture system to facilitate study of this regulation.Acinar cells were isolated by enzymatic disaggregation from pig tracheal submucosal tissue and cultured on a plastic substrate coated with human placental collagen. The fine structure of the cells in confluent culture was studied by conventional transmission electron microscopy. The elemental content in resting cells and stimulated cells grown on a permeable substrate was studied by X-ray microanalysis.The cultured cells retained structural characteristics (microvilli, secretory granules and desmosomes) of in situ epithelia. The total intracellular Cl and K concentrations significantly decreased after stimulation with the cholinergic agonist acetylcholine, the predominantly alpha-adrenergic agonist norepinephrine, or the beta-adrenergic agonist isoproterenol. Both ionomycin and 8-bromoadenosine 3':5'-cyclic monophosphate (8-bromo-cAMP) caused a marked decrease of the intracellular Cl, K and Na concentrations. Cl -secretion induced by acetylcholine was inhibited by Cl -channel blockers anthracene-9-carboxylic acid and 5-nitro-2-(3-phenylpropyl-amino)-benzoic acid (NPPB), but Cl -efflux induced by 8-bromocAMP was blocked only by NPPB. The intracellular Cl and Na content significantly increased and the cellular K content markedly decreased after treatment with ouabain.These results indicate that the cultured acinar cells maintained the structural and functional characteristics of in situ tissue and that this system is suitable for studying aspects of ion and water transport by the airway submucosal gland cells.

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Ion transport in primary cultures from human sweat gland coils studied with X‐ray microanalysis

Cited by 16 publications

References 12 publications

Regulation of Ion Content in Primary Cultures from Reabsorptive Ducts of Human Sweat Glands Studied by X-ray Microanalysis.

Regulation of Ion Content in Primary Cultures from Reabsorptive Ducts of Human Sweat Glands Studied by X-ray Microanalysis.

Activation of DeltaF508 CFTR in a cystic fibrosis respiratory epithelial cell line by 4-phenylbutyrate, genistein and CPX

Ion transport in cultured pig tracheal submucosal gland acinar cells studied by X-ray microanalysis

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