Search PubMed⌕ Search

Biomedical subjects

P S Pedersen

Publications and source records attributed to P S Pedersen.

At least 19 recordsLinked to original sources
PubMed2000

The sarco/endoplasmic reticulum calcium-ATPase 2b is an endoplasmic reticulum stress-inducible protein.

The sarco/endoplasmic reticulum calcium-ATPase (SERCA) translocates Ca(2+) from the cytosol to the lumen of the endoplasmic reticulum. This Ca(2+) storage is important for cellular processes such as calcium signaling and endoplasmic reticulum (ER)-associated posttranslational protein modifications. We investigated the expression of the SERCA2 and SERCA3 isozymes in PC12 cells exposed to agents interfering with different aspects of the posttranslational protein processing within the ER, thereby activating the ER stress-induced unfolded protein response (UPR). All agents increased the SERCA2b mRNA level 3-4-fold, in parallel with increasing mRNA levels for the ER stress marker proteins BiP/GRP78 and CHOP/GADD153. In contrast, SERCA3 mRNA levels did not change. SERCA2b mRNA stability was not changed, indicating that the mechanism of its up-regulation was transcriptional, in accordance with the presence of ER stress response elements in the promoter region of the SERCA2 gene. SERCA2b was also increased at the protein level upon ER stress treatments. Induction of ER stress by tunicamycin, dithiothreitol, or l-azetidine 2-carboxylic acid did not result in depletion of ER calcium, showing that such depletion was not necessary for up-regulation of SERCA2b expression or UPR activation in general. We conclude that the SERCA2b expression can be controlled by the UPR pathway independently of ER Ca(2+) depletion.

Animals↗
PubMed1999

Ion transport in epithelial spheroids derived from human airway cells.

In the present study, we describe a novel three-dimensional airway epithelial explant preparation and demonstrate its use for ion transport studies by electrophysiological technique. Suspension cultures of sheets of epithelial cells released by protease treatment from cystic fibrosis (CF) and non-CF nasal polyps developed free-floating, monolayered epithelial spheres, with the apical, ciliated cell membrane facing the bath and the basolateral cell membrane pointing toward a fluid-filled lumen. Microelectrode impalement of both non-CF and CF spheroids revealed lumen-positive transepithelial electrical potential differences (PDs) that were inhibited by amiloride, indicating that the spheroids were inflated due to amiloride-sensitive Na+ absorption followed by water. Transformation to a Cl- secretory state was achieved by addition of ATP to the bath, leading to the development of a diphenylamine-2-carboxylate-sensitive PD. A cAMP-induced increase in PD was seen in non-CF spheroids only. In response to hydrocortisone treatment, Na+ transport reflected by amiloride-sensitive PD increased and more so in CF than in non-CF spheres. We concluded that this preparation is a useful model for the airway surface epithelium and is suitable for studies of transport mechanisms and regulation.

Absorption↗
PubMed1999

Fluid absorption related to ion transport in human airway epithelial spheroids.

Airway epithelium explants from cystic fibrosis (CF) patients and non-CF subjects formed monolayered spheres, with the apical ciliated cell membrane facing the bath and the basolateral cell membrane pointing toward a fluid-filled lumen. With the use of two microelectrodes, transepithelial potential difference and changes in potential difference in response to passage of current pulses were recorded, and epithelial resistance and the equivalent short-circuit current were calculated. Non-CF control potential difference and short-circuit current values were significantly lower than the CF values, and amiloride inhibited both values. Fluid transport rates were calculated from repeated measurements of spheroid diameters. The results showed that 1) non-CF and CF spheroids absorbed fluid at identical rates (4.4 microl x cm(-2) x h(-1)), 2) amiloride inhibited fluid absorption to a lower residual level in non-CF than in CF spheroids, 3) Cl(-)-channel inhibitors increased fluid absorption in amiloride-treated non-CF spheroids to a level equal to that of amiloride-treated CF spheroids, 4) hydrochlorothiazide reduced the amiloride-insensitive fluid absorption in both non-CF and CF spheroids, and 5) osmotic water permeabilities were equal in non-CF and CF spheroids ( approximately 27 x 10(-7) cm x s(-1) x atm(-1)).

Amiloride↗
PubMed1992

Chloride and potassium conductances of cultured human sweat ducts.

The purpose of this study was to characterize the ion conductances, in particular those for Cl- and K+, of human sweat duct cells grown in primary culture. Sweat duct cells from healthy individuals were grown to confluence on a dialysis membrane, which was then mounted in a mini-Ussing chamber and transepithelial and intracellular potentials were measured under open-circuit conditions. Under control conditions the epithelia developed mucosa-negative transepithelial potentials, Vte, of about -10 mV. The apical membrane potential, Va, was -25 mV to -30 mV (n = 97) in most cells, but several cells had a higher potential of about -55 mV (n = 29). Mucosal amiloride (10 mumol/l) hyperpolarized Va from -31 +/- 1 mV to a new sustained level of -46 +/- 2 mV (n = 36). These changes were accompanied by increase in the fractional resistance of the apical membrane, fRa, and decreases of Vte and the equivalent short-circuit current, Isc. In amiloride-treated tissues an increase in mucosal K+ concentration (5 mmol/l to 25 mmol/l) depolarized Va by 5 +/- 1 mV (n = 8), while the same step on the serosal side depolarized Va by 20 +/- 2 mV (n = 8). A Cl- channel blocker 3',5-dichlorodiphenylamine-2-carboxylate DCl-DPC; 10 mumol/l) depolarized Va by 5 +/- 1 mV (n = 6), an effect that was lost after amiloride application. The blocker had no effect from the serosal side. Reduction of mucosal Cl- (from 120 to 30 or 10 mmol/l) depolarized Va by 9-11 mV (n = 35), an effect that was often followed by a secondary hyperpolarization of 10-30 mV (n = 27).(ABSTRACT TRUNCATED AT 250 WORDS)

Amiloride↗
PubMed1992

Influence of extracellular bicarbonate on the short-circuit current and intracellular free calcium of human cultured sweat duct cells.

Transepithelial short-circuit current (Iscc) and intracellular free Ca2+ (Ca2+i) was studied in monolayers of cultured human sweat duct cells (CSDCs) in the presence or absence of HCO3- (and CO2) in the bathing solutions. Addition of HCO3- (and CO2) increased the control Iscc by more than 50%. The effect of HCO3- (and CO2) on Iscc was confined to the serosal bath. The HCO3- (and CO2) effect was also studied during stimulation with the cholinergic agonist methacholine (MCh), which in CSDC induces a complex response consisting of an initial Iscc and Ca2+i spike, which is independent of extracellular Ca2+, followed by regular Iscc and Ca2+i oscillations, which are absent during Ca(2+)-free bathing conditions. The sustained Iscc and Ca2+i oscillations, but not the initial Iscc and Ca2+i spike were abolished by the removal of extracellular HCO3- (and CO2). It is concluded that the Ca2+ influx and the Iscc in CSDCs are critically influenced by the presence of extracellular HCO3- (and CO2) in the bathing solutions.

Amiloride↗
PubMed1991

Separate agonist-specific oscillatory mechanisms in cultured human sweat duct cells.

1. Cultured sweat duct cells (CSDCs) were grown to confluency on a permeable support, and the pharmacological ion transport regulation was assayed by transepithelial voltage clamp techniques. 2. Exposure of the serosal membrane of CSDCs to methacholine (MCh), lysylbradykinin (LBK) or histamine produced an oscillating short-circuit current (Iscc) response, which could be divided in an initial transient phase and a sustained oscillating phase, the latter of which was totally dependent on external Ca2+. 3. The Iscc responses evoked by LBK and histamine were, in contrast to the cholinergic response, characterized by a marked desensitization and short duration of the subsequent phase of Iscc oscillations. 4. Prolonged Iscc oscillations, reflecting continuous Ca2+ influx, were seen following MCh stimulation, and in response to LBK or histamine stimulation, when cells had been pre-treated with MCh. This pre-treatment effect of MCh was independent of continuous muscarinic receptor occupation, and it was unrelated to nicotinic receptor occupation. 5. It is suggested that MCh stimulation selectively initiates an influx of Ca2+ to an intracellular pool, from where Ca2+ can be discharged repetitively. In contrast, LBK and histamine only activate discharge of Ca2+ from such an intracellular pool, resulting in a limited response, given no prior stimulation by MCh of the Ca2+ influx mechanism.

Calcium↗
PubMed1990

Cholinergic-induced oscillating transepithelial short-circuit current in cultured human sweat duct cells.

Human sweat duct cells in primary culture were investigated by voltage-clamp technique. Stimulation with the muscarinic agonist, metacholine (MCh), produced an abrupt transient rise followed by sustained regular oscillations in the transepithelial short-circuit current (Iscc), which in these cells is carried by a mucosal amiloride-sensitive Na+ influx, secondary to a Ca2(+)-activated, voltage-dependent, large K+ shunt across the serosal membrane. The time of latency, the initial transient phase, and the sustained oscillating phase of the MCh-induced Iscc response were demonstrated to be differently affected by changes in temperature, agonist concentration and external Ca2+ supply. From these results a model is proposed for the MCh-induced signal transduction in cultured sweat duct cells, involving a primary intracellular oscillatory Ca2+ mobilization, activated by IPP, sustained by a temperature-regulated external Ca2+ supply, and counter-regulated by cytosolic Ca2+.

Calcium↗
PubMed1990

Chloride permeability regulation via a cyclic AMP pathway in cultured human sweat duct cells.

1. Isolated coiled reabsorptive sweat ducts from normal subjects and patients with cystic fibrosis (CF) were cultured in vitro. Cells were harvested and plated onto permeable supports to form confluent cell sheets. The Ussing chamber technique was used to study pharmacological regulation of the transepithelial ion transport in these membranes. 2. Addition of a stable cyclic AMP analogue, 8-Br-cyclic AMP, to normal cell cultures resulted in a decrease of the transepithelial potential difference (PD). 3. Forskolin exposure resulted in a similar PD decrease, which was augmented by the phosphodiesterase inhibitor, isobutylmethylxanthine (IBMX). 4. Exposure to isoprenaline, prostaglandin E2 (PGE2), and phenylephrine resulted in a response mimicking the forskolin-induced response, that was also amplified by IBMX. 5. Pre-incubation with cholera toxin abolished the isoprenaline response and reduced the control resistance. 6. Propranolol abolished the responses induced by isoprenaline and phenylephrine, whereas phentolamine had no effect. PGE2-induced responses were inert to both types of blockers. 7. Indomethazine addition to an unstimulated membrane resulted in a weak PD increase, i.e. a response opposite to that induced by isoprenaline. 8. IBMX addition to an unstimulated membrane resulted in a weak isoprenaline-like response. When the cells were pre-treated with indomethazine this IBMX response was absent. 9. Unidirectional Cl- isotope flux studies demonstrated a large increase of net Cl- reabsorption in response to isoprenaline and PGE2. 10. Mannitol isotope flux studies revealed that the paracellular permeability was unaffected by isoprenaline exposure. 11. Membranes derived from CF patients did not respond similarly to any of these agents. However, a weak spike, occasionally followed by a gradual increase of the short-circuit current (Iscc), was observed in both normal subjects and CF patients. 12. It is concluded that the primary effect on ion transport of factors increasing the cyclic AMP in normal cultured sweat duct cells is an activation of a transcellular Cl- permeability. This effect was missing in cells derived from CF patients.

1-Methyl-3-isobutylxanthine↗
PubMed1990

Cation transport by sweat ducts in primary culture. Ionic mechanism of cholinergically evoked current oscillations.

1. The coiled reabsorptive segment of human sweat ducts was cultured in vitro. Cells were then harvested and plated onto a dialysis membrane which was glued over a hole in a small disc. Cultures were maintained in a low serum, hormone-supplemented medium that allowed the cells to grow to confluency. The disc was then placed as a partition between two compartments of a miniature Ussing chamber. The chamber was mounted on the stage of an inverted microscope and intracellular potentials were recorded under transepithelial open-circuit or voltage clamp conditions. All values are given as means +/- S.E.M. and n refers to the number of preparations or duct cells. 2. Under control conditions, the cultured epithelia developed mucosa-negative transepithelial potentials (Vt) ranging from -2.5 to -38 mV (-13.5 +/- 1.5 mV, n = 36). The basolateral membrane potential (Vb) was -39.4 +/- 0.7 mV (n = 50 cells), and the apical membrane potential (Va) was linearly correlated with Vt:Va = 1.0 Vt -39.3 mV (r = -0.78, n = 50). 3. The epithelium generated inwardly directed short-circuit currents (Isc) of 12-95 microA cm-2 (45 +/- 4 microA cm-2, n = 36) with a steady-state intracellular potential. Vc = -31.1 +/- 0.6 mV and a fractional resistance of the apical membrane, fR = 0.59 +/- 0.01 (n = 115 cells). 4. The Na+ channel blocker amiloride (mucosal bath, 10 microM) abolished Isc -0.8 +/- 0.6 microA cm-2), the cells hyperpolarized to -61.0 +/- 1.2 mV, and fR increased to 0.85 +/- 0.01 (n = 44). These effects were fully reversible. 5. During initial stimulation with the cholinergic agonist, methacholine (serosa, 5 or 10 microM), the short-circuit current increased to 80 +/- 10 microA cm-2, the cells hyperpolarized to -55.8 +/- 1.2 mV, and fR increased to 0.82 +/- 0.01 (n = 35). 6. In short-circuited preparations stimulated with methacholine an increase in mucosal potassium concentration ([K+]m) from 5 to 25 mM had no significant effect, while a similar increase in the serosal K+ concentration ([K+]s) produced a change in Vc of 44 +/- 3 mV per log10[K+]s (n = 9). In non-stimulated preparations this change was only 16 +/- 2 mV per log10[K+]s (n = 13). After blocking the apical Na+ channels with amiloride the slope was 24 +/- 5 mV per log10[K+]s in unstimulated preparations.(ABSTRACT TRUNCATED AT 400 WORDS)

Amiloride↗
PubMed1989

Formation of inositol polyphosphates in cultured human sweat duct cells in response to cholinergic stimulation.

Inositol phosphate formation in response to cholinergic stimulation was studied in cultured human sweat duct cells, prelabelled with myo-[2-3H]inositol. Formation of inositol mono-, bis-, tris- and tetrakisphosphates was increased after 15 min stimulation by 30 microM carbachol. Formation of inositol 1,3,4-trisphosphate and inositol tetrakisphosphate was significantly increased within 1 min at carbachol concentrations between 10 microM and 100 microM. No detectable increase in inositol 1,4,5-trisphosphate formation was observed at 15 s or 1 min, but an increase was observed after 15 min at a carbachol concentration of 30-100 microM. The data are consistent with an involvement of inositol polyphosphates in the biphasic response of ion transport, to cholinergic stimulation in these cells (see Pederson, P.S. (1986) 6th Professional Conference "Broken Arrow 1986". Genetic and Eptihelial Dysfunction in Cystic Fibrosis (Riordan, J.R. and Buchwalds, M., eds.), Alan Liss, New York and Pedersen, P.S. (1987) Med. Sci. Res. 15, 769-770) and suggest a different pattern of metabolism from exocrine acinar cells.

Calcium↗
PubMed1989

Human sweat duct cells in primary culture. Basic bioelectric properties of cultures derived from normals and patients with cystic fibrosis.

Human sweat duct cells from the coiled reabsorptive segment have been cultured successfully, free from fibroblasts, in a low serum, hormone-supplemented medium, Ham's F12. The cultured cells exhibited a typical epithelial cobblestone pattern, and microvilli-covered luminal cells were seen joined together with typical junctional complexes. In cultures derived from normals and patients with cystic fibrosis (CF), growth and morphologic characteristics were indistinguishable. When grown on a membranous support, and mounted in an Ussing chamber, vectorial electroconductive ion-transport could be identified. The epithelial preparations produced active mucosa to serosa-directed sodium flux via amiloride-sensitive, apical sodium channels and ouabain-sensitive sodium pumps located in the basolateral membrane, which also contained a potassium shunt. These findings are consistent with a polarized epithelium with properties similar to the intact organ. High transepithelial resistance and increased amiloride sensitivity were typical for cells derived from CF, indicating that principal normal as well as pathologic properties of the sweat duct are preserved in culture.

Adolescent↗
PubMed1988

Isolation of a human gene with protein sequence similarity to human and murine int-1 and the Drosophila segment polarity mutant wingless.

An expressed gene sequence which was identified by the isolation of a methylation free CpG island from human chromosome 7 has been cloned from a human lung cDNA library. The deduced protein sequence contains 360 amino acids and has several features of a secreted protein; it is cysteine rich with a signal peptide sequence and two potential asn-linked glycosylation sites. The protein sequence shows marked similarity with human and murine int-1 and their Drosophila homolog wingless (Dint-1). This human int-1 related protein, int-1 and Dint-1 have diverse patterns of expression, but the inferred structural similarities suggest that some of the functional characteristics of these proteins may be shared.

Amino Acid Sequence↗
PubMed1985

Desamino-D-arginine vasopressin in childhood nocturnal enuresis.

The effect of desamino-D-arginine vasopressin was investigated in a double-blind study of 37 children more than 9 years old with nocturnal enuresis resistant to conventional therapy. A significant reduction of wet nights was observed but as soon as the medication was stopped the children reverted to earlier bedwetting habits.

Adolescent↗
PubMed1984

Methylprednisolone resistance of cystic fibrosis lymphocytes.

In the current study, the in vitro viability and resistance to methylprednisolone (MP) was investigated in lymphocytes from patients with cystic fibrosis (CF) and normal controls. The addition of MP to phytohemagglutinin-stimulated lymphocyte cultures inhibits the proliferative response in a dose-dependent way. The dose response curves to MP were similar in CF cultures and controls (P greater than 0.05). Neither preincubation nor pulse exposure to MP revealed any difference between normal and CF lymphocytes. Skin fibroblasts from patients with CF have been reported to show increased resistance to the toxic effect of dexamethasone (1-3, 5-6). This study was undertaken to investigate the suppressive effect of glucocorticoids on another cell culture system using MP on phytohemagglutinin-stimulated lymphocytes.

Adolescent↗