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  • 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
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    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
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 3
    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
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 4
    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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  • 5
    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
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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