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  • Adrenalectomy  (5)
  • Cell culture  (5)
  • Cl−/HCO 3 − exchange  (2)
  • 1
    Digitale Medien
    Digitale Medien
    Springer
    Pflügers Archiv 405 (1985), S. S167 
    ISSN: 1432-2013
    Schlagwort(e): Corneal endothelium ; Cell culture ; Intracellular potential ; Sodium-bicarbonate cotransport ; pH regulation ; Stilbenes
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Intracellular potential measurements on confluent monolayers of cultured bovine corneal endothelial cells were used to define passive ion transport processes in these cells. Previous studies [11, 12] have provided the experimental basis for a cellular model, is which bicarbonate entry across the basolateral membrane in indirectly driven by a Na+/H+-exchanger, which is inhibitable by amiloride (1 mmol/l). Na+ and HCO 3 − leave the cell via an electrogenic bicarbonate sodium cotransport, which is inhibitable by the disulfonic stilbene derivates SITS or DIDS. This model is also compatible with transepithelial work from other groups. In this paper, we briefly review the evidence we have obtained for this model.
    Materialart: Digitale Medien
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  • 2
    ISSN: 1432-2013
    Schlagwort(e): Tubular Fluid Reabsorption ; Water Permeability ; Adrenalectomy ; Adrenal Cortical Hormones ; Glucocorticoids ; Tubuläre Flüssigkeitsresorption ; Wasserpermeabilität ; Adrenalektomie ; Nebennierenrindenhormone ; Glucocorticoide
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary In order to analyse the mechanism of inhibition of water diuresis in adrenal insufficiency renal surface tubulus of male albino rats (group I: controls, group II: 8 days or more after bilateral adrenalectomy) have been perfused using the free flow microperfusion technique. From the percent osmotic equilibration of a hypotonic perfusate as measured cryoscopically osmotic water permeability (L p) has been calculated. Furthermore, percent osmotic equilibration has been estimated from transtubular net water flux measured with 14C-inulin. With both methods which constitute a maximum estimate (cryoscopic data) and a minimum estimate (inulin data) osmotic water permeability was found to be increased after adrenalectomy in distal convolutions. Hormone substitution with either cortisone (2.5 mg/100 g b.w. 24 hours i.m.) or dexamethasone (0.05 mg/100 g b.w. 24 hours i.m.) reestablished towards normal water permeability after 3 days of treatment. The data are compatible with the concept of a direct effect of glucocorticosteroids on water permeability of distal tubule membranes. In the proximal convolution osmotic water permeability was unaffected by bilateral adrenalectomy.
    Materialart: Digitale Medien
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  • 3
    ISSN: 1432-2013
    Schlagwort(e): Split-Oil Droplet Method ; Proximal and Distal Convolution ; Ultrastructure of Kidney ; Na- and Fluid Reabsorption ; Adrenalectomy ; Öltropfenmethode ; proximales und distales Tubuluskonvolut ; Ultrastruktur der Niere ; Natrium- und Flüssigkeitsresorption ; Adrenalektomie
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Micropuncture experiments have been carried out in proximal and distal convolutions of the rat kidney in order to test the reliability of the split-oil droplet method. 1. Light- and electronmicroscopic results: In castor oil filled segments a sufficient blockade is achieved, as judged from the compression of the brush border. In the test segments, interposed between two oil filled segments which are injected with ferritin containing saline, the tubular epithelium and especially the luminal brush border exhibit a normal histology. The tight junctions are intact and pinocytosis is indicated by ferritin resorption. Only in very few instances were oil droplets found to adhere to the brush border of the test segment, sometimes forming circumscript deposits protruding towards the cells. In such areas mechanical lesions of the cell membrane were only very rarely observed. In the proximal and distal convolutions of adrenalectomized rats the same experimental procedure did not lead to differences in the morphology of the epithelium. 2. Physiological data: a) Half time of volume reabsorption was unchanged when the split-oil droplet procedure was applied repeatedly to the same tubular segment. b) Intraluminal application of KCN (10−3–10−2 Mol/l) inhibited reabsorption of NaCl and fluid out of the test segment. c) Addition of the non-permeating polyethyleneglycol to the test solution prolongedt1/2 to infinity. d) In experimental diabetes insipidus half time of fluid reabsorption was greatly increased in the distal convolution, but could be normalized by intravenous injection of vasopressin. This was true even when the tubular epithelium was in contact with castor oil for more than 60 min prior to the administration of the hormone. From the results obtained in this and earlier studies it is concluded that contact with castor oil and tubular dilatation (to the extent that occurs during the procedure of the split-oil droplet method) does not damage tubular epithelium to a measureable degree as judged both by functional and morphological criteria. This holds also for adrenalectomized rats.
    Materialart: Digitale Medien
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  • 4
    Digitale Medien
    Digitale Medien
    Springer
    Pflügers Archiv 347 (1974), S. 117-123 
    ISSN: 1432-2013
    Schlagwort(e): Potassium Selective Liquid Ion-Exchange Microelectrode ; Intracellular Potassium Concentration ; Distal Tubule Cell Potassium ; Aldosterone ; Adrenalectomy
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Using a double-barreled K+-selective liquid ion-exchange microelectrode effective intracellular potassium concentration and peritubular membrane potential were measured simultaneously in single cells of the distal tubule of the rat kidney. A mean K-intracellular of 47.2±1.8 mMol/l was obtained in control rats. The effective K-intracellular decreased significantly to 39.1±1.4 mMol/l after adrenalectomy. Acute administration of aldosterone to adrenalectomized rats (15 μg/100 g B.W. twice a day for 1–2 days) was without effect on K-intracellular. Substitution with aldosterone over a period of 5 days normalized the effective intracellular potassium concentration to 45.2±0.9 mMol/l. The peritubular membrane potential increased. Aldosterone regulates potassium secretion by at least two mechanisms: In addition to the already demonstrated effect on passive potassium permeability of the luminal cell membrane, aldosterone stimulates the active uptake of potassium at the peritubular membrane, thereby regulating the effective intracellular potassium concentration. Both mechanisms increase the effective driving force for passive entry of potassium into the distal tubular lumen.
    Materialart: Digitale Medien
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  • 5
    Digitale Medien
    Digitale Medien
    Springer
    Pflügers Archiv 345 (1973), S. 159-178 
    ISSN: 1432-2013
    Schlagwort(e): Micropuncture ; Transepithelial Potential ; Peritubular Membrane Potential ; Partial Potassium Conductance ; Adrenalectomy ; Aldosterone
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Electrophysiological studies were performed in proximal and distal tubules of rat kidneys. To assess the relative potassium permeability the tubular lumen was perfused with solutions of varying K concentrations while the transepithelial PD were measured. In the distal tubule the transport number for potassium was 0.43 in control rats and 0.27 in adrenal insufficiency. Acute (5 μg/100 g B.W.) or chronic (30 μg/100 g B.W. daily for 3 days) aldosterone administration normalized the relative permeability. This effect of aldosterone could not be prevented by cycloheximide (100 μg/100 g B.W.). The peritubular membrane potential of the distal tubule decreased from 75 mV in control rats to 62 mV in adrenalectomized rats. Acute injection of aldosterone had no effect on peritubular membrane potential. In the proximal tubule qualitatively similar results were obtained as in the distal tubule. The impairment of potassium secretion in adrenal insufficiency is at least partly due to a reduced permeability of the luminal membrane of the distale tubule. Aldosterone acutely stimulates potassium secretion by increasing the luminal permeability of the distal tubule. In comparison to the effect of aldosterone on active transport components the increase in luminal potassium permeability by aldosterone may be either less sensitive to blockade by inhibitors of protein synthesis or is not dependent upon induction of newly synthetized proteins.
    Materialart: Digitale Medien
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  • 6
    Digitale Medien
    Digitale Medien
    Springer
    Pflügers Archiv 348 (1974), S. 155-165 
    ISSN: 1432-2013
    Schlagwort(e): Transepithelial Potential ; Sodium Conductance ; Adrenalectomy ; Aldosterone ; Cycloheximide
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary To asses passive permeability properties of distal, and proximal tubules of the rat kidney the tubular lumen was perfused with solutions of 1.5 and 150 mM Na/l while transtubular potential differences were recorded. Sodium transport numbers (T Na) were calculated.T Na in the distal tubule of adrenalectomized rats was acutely increased from 0.21 to 0.27 by aldosterone (5 μg/100 g B.W.). This effect of aldosterone could not be reduced by concomitant injection of cycloheximide (100 μg/100 g B.W.). Aldosterone was also effective in control rats. In the proximal tubule similar data were obtained. However, the aldosterone-induced increase of conductance was slightly reduced with cycloheximide. These measurements of transepithelial sodium conductance indicate that aldosterone, in addition to the already known stimulation of active sodium transport, increases overall permeability of the tubular wall to sodium. In the distal tubule this effect indicates an increase of the luminal membrane permeability whereas in the proximal tubule aldosterone may facilitate the diffusion of sodium through the intercellular shunt path and/or the luminal membrane. The passive components of transepithelial electrolyte transfer seem to be less sensitive to inhibition of protein synthesis than the active transport components.
    Materialart: Digitale Medien
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  • 7
    ISSN: 1432-2013
    Schlagwort(e): Ciliary muscle ; Cell culture ; Intracellular calcium ; Isolated ciliary muscle strips ; Contractility ; Acetylcholine ; Carbachol
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Electromechanical and pharmacomechanical coupling was investigated in human ciliary muscle by measuring the intracellular free calcium in single cultured ciliary muscle cells and the contractility in meridional ciliary muscle strips. The basal resting calcium concentration was 75±8.7 nmol/l, n=23. Application of acetylcholine (0.1 mmol/l) and carbachol (0.1 mmol/l) resulted in an initial [Ca2+]i peak followed by a recovery phase and a [Ca2+]i plateau. The initial [Ca2+]i peak was still observed in the absence of extracellular calcium and in the presence of verapamil (0.1 mmol/l). During its plateau [Ca2+]i was decreased by withdrawal of extracellular calcium or application of verapamil (0.1 mmol/l). Depolarization induced by a high level of extracellular potassium yielded only a small transient [Ca2+]i peak without a [Ca2+]i plateau. In isolated ciliary muscle strips, muscarinic stimulation (carbachol 0.1 mmol/l) resulted in an initial phasic and a subsequent tonic contraction. Removal of external calcium reduced the phasic contraction to 30.6±4.4% (n=8) and completely abolished the tonic one. Verapamil (0.1 mmol/l) had only a slight relaxing effect when applied during the tonic contraction. We conclude that human ciliary muscle contraction is mediated by calcium release from intracellular stores and calcium entry through calcium channels, which are most probably receptor-operated. Depolarization of the muscle cell membrane and calcium entry through voltage-operated calcium channels do not contribute significantly to human ciliary muscle contraction.
    Materialart: Digitale Medien
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  • 8
    ISSN: 1432-2013
    Schlagwort(e): Human ciliary muscle ; Smooth muscle ; Cell culture ; Intracellular calcium ; Membrane potential ; Acetylcholine ; Endothelin
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract We characterized the effects of acetylcholine and endothelin on cultured human ciliary muscle cells, using the calcium-sensitive dye fura-2 to measure intracellular calcium and intracellular microelectrodes to measure the membrane potential. Both agonists, endothelin and acetylcholine, had a typcial biphasic effect on the intracellular calcium concentration. Calcium peaked initially, because of its release from intracellular stores, and then reached a plateau, owing to entry of extracellular calcium. Endothelin-induced calcium entry was almost completely blocked by addition of extracellular La3+ (50 μmol/l) and Ni2+ (1 mmol/l). Acetylcholine-induced calcium entry was likewise almost completely abolished by La3+ and Ni2+. Both endothelin and acetylcholine led to an initial transient hyperpolarization with a subsequent depolarization. The hyperpolarization of the membrane potential had a time course similar to the initial calcium peak, while the depolarization occurred parallel to the calcium plateau. The depolarization induced by both agonists was reduced in the presence of La3+ and Ni2+. Verapamil (10 μmol/l) had no effect on either the calcium entry or the depolarization. Acetylcholine did not induce a [Ca2+]i peak when it was applied during the endothelin-induced [Ca2+]i plateau and vice versa. The [Ca2+]i plateau was not higher with concomitant than with single application of acetylcholine or endothelin. Thus, calcium entry and membrane depolarization induced by acetylcholine and endothelin seem to be mediated by a common La3+- and Ni2+-sensitive but verapamil-insensitive mechanism.
    Materialart: Digitale Medien
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  • 9
    ISSN: 1432-2013
    Schlagwort(e): Bicarbonate ; Intracellular potentials ; Bicarbonate sodium cotransport ; Chloride ; Anion exchanger ; Cell culture ; Cornea ; Endothelium ; Furosemide ; Barium
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Using intracellular microelectrode technique, the effect of anion substitution on the voltage responses to extracellular bicarbonate and sodium was explored in cultured bovine corneal endothelial cells. 1. The overall amplitude of voltage changes induced by periodic changes of [HCO 3 − ]0 (depolarization upon removal of HCO 3 − and hyperpolarization upon readdition) was reduced when Cl− was replaced by organic anions (cyclamate, methylsulfate, benzenesulfonate) or by SO 4 2− , and to a lesser extent by substitution with Br−. 2. There was a similar effect of anion substitution on the response to changes of [Na+]0. 3. In both cases, in the absence of Cl, the voltage V returned at a slower rate to baseline levels after it had been transiently changed by either an imposed Na- or HCO3-gradient, indicating a slower dissipation of these gradients. The direct response of V to these imposed gradients was affected only to a minor degree. 4. Replacement of Cl− by SO 4 2− or organic anions led to a slow, reversible depolarization of the cell, while substitution with Br− had only a slight effect. 5. The effect of anion substitution on the voltage responses to HCO 3 − or Na+ could not be mimicked by a depolarization induced by Ba2+ (1 mM). 6. Furosemide (10−3 M) led to a slight reduction of the voltage responses to HCO 3 − , but could not suppress the effect of anion substitution on these reactions. It could neither suppress the depolarization induced by anion substitution and had no effect on steady-state PD. 7. It is suggested, that cultured bovine corneal endothelial cells, in addition to a previously demonstrated electrogenic HCO 3 − −Na+-cotransport, which is probably not dependent on Cl, possess an electroneutral mechanism for HCO 3 − and/or Na+-movement, which depends on Cl. No evidence for a Cl-conductance could be obtained.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 10
    ISSN: 1432-2013
    Schlagwort(e): Cell culture ; pH sensitive dyes ; pH sensitive absorbance ; 5 (and 6)-carboxy-dimethylfluorescein ; Na+/H+ antiport ; Cl−/HCO 3 − exchange
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Regulation of intracellular pH (pHi) in bovine retinal pigment epithelium (RPE) was investigated in cell culture. pHi was measured using the pH-sensitive absorbance of intracellularly trapped 5 (and 6)-carboxy-dimethyl-fluorescein (CDMF). (1) Regulation of pHi after induction of an acid load by removal of NH4Cl could be blocked either totally by removal of extracellular sodium, or subtotally (about 90%) by application of amiloride (1 mmol/l). Additional flux measurements revealed a dose-dependent, amiloride-sensitive22Na+-uptake into Na+-loaded cells. Both results suggest the presence of a Na+/H+ antiport. (2) When alkalinization of the cells was induced by preincubation with 50 mmol/l acetate in HCO 3 − -Ringer's and subsequent removal of the weak acid, the following regulation was dependent on the presence of extracellular chloride. This process could be blocked with DIDS (1 mmol/l), suggesting the presence of a Cl−/HCO 3 − exchange mechanism. (3) We found no evidence for a Na+/HCO 3 − -cotransport, which had been postulated to be present in RPE by others. We conclude that two processes are involved in regulation of pHi in RPE: A Na+/H+ antiport responsible for recovery of pHi from acid load, and a DIDS-sensitive Cl−/HCO 3 − exchange mechanism responsible for recovery of pHi after alkalinization.
    Materialart: Digitale Medien
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