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  • 1
    Digitale Medien
    Digitale Medien
    Voltage clamping of Xenopus laevis oocytes utilizing agarose-cushion electrodes (1994)
    Springer
    Pflügers Archiv 426 (1994), S. 453-458 
    Details
    ISSN: 1432-2013
    Schlagwort(e): Agarose-cushion electrodes ; Charging compensator ; Two-electrode voltage clamp ; Xenopus laevis oocytes ; Ion channel modulation ; Heterologous expression
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Two-electrode voltage clamping of expressed ion channels in intact oocytes of the South African clawed frog Xenopus laevis has been refined to allow stable, low-resistance electrical access to the cytosol (50–800 kΩ). Glass microelectrodes were filled with a cushion of 1 % agarose at their tips to prevent KC1 leakage (agarose-cushion electrodes). Insertion of these electrodes into X. laevis oocytes yielded stable preparations for periods of more than l h with a stable input resistance of 1–4MΩ. Furthermore, a simple modification of the voltage-clamp circuit (charging compensator) is described that increases the flexibility of arrangements for differential recording of the membrane potential in order to subtract voltage drops across a series resistance. The result is a considerable increase in the practically attainable speed of the voltage clamp with the conventional two-electrode arrangement. The performance of the charging compensator was tested on an equivalent circuit that simulates the oocyte and electrodes. In addition, the combination of agarose-cushion electrodes and the charging compensator was tested on oocytes expressing Shaker H4 currents. The fidelity of the voltageclamp circuit was also verified by measuring the membrane potential with additional independent microelectrodes connected to a differential amplifier, independent of the two-electrode voltage clamp system. The system described here will be useful for ion channel studies in X. laevis oocytes requiring long-term recordings and/or measurements of large, fast ion currents.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00388310
    Standort Signatur Einschränkungen Verfügbarkeit
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    Artikel (Nationallizenzen)
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  • 2
    Digitale Medien
    Digitale Medien
    Serotonin and protein kinase C modulation of a rat brain inwardly rectifying K+ channel expressed inXenopus oocytes (1996)
    Springer
    Pflügers Archiv 431 (1996), S. 335-340 
    Details
    ISSN: 1432-2013
    Schlagwort(e): Serotonin ; Inward rectifying potassium channel ; Brain ; PKC ; 5-HT2C receptor ; Heterologous expression ; Xenopus laevis oocytes ; Modulation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract InXenopus laevis oocytes injected with rat brain poly(A)+ RNA, perfusion with a high-K+ solution (96 mM KCl) generated an inward current (I HK) which was absent in water-injected oocytes. Part ofI HK was blocked by low concentrations of Ba2+ (half-maximal inhibitory concentration, IC50: 4.2 ± 0.5 μM). When serotonin (5-HT) was applied to these oocytes a transient inward oscillating Cl− current arising from activation of Ca2+ -dependent Cl− channels,I Cl(Ca), was observed. When this response decayed, a 30% reduction ofI HK could be detected. Electrophysiological characterization of the K+ channel down-modulated by 5-HT revealed that it is an inward rectifier. Antisense suppression experiments revealed that the 5-HT2C receptor mediates the down-modulatory effect of 5-HT. The nature of the modulatory pathway was investigated by application of phorbol esters and intracellular injection of protein kinase C (PKC) inhibitors, ethylenebis (oxonitrilo)tetraacetate (EGTA) and inositol 1, 4, 5-trisphosphate. The results demonstrate that PKC is responsible for the down-modulatory effect.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF02207270
    Standort Signatur Einschränkungen Verfügbarkeit
    BibTip Andere fanden auch interessant ...
    Artikel (Nationallizenzen)
    Volltext
  • 3
    Digitale Medien
    Digitale Medien
    Serotonin and protein kinase C modulation of a rat brain inwardly rectifying K+ channel expressed in Xenopus oocytes (1996)
    Springer
    Pflügers Archiv 431 (1996), S. 335-340 
    Details
    ISSN: 1432-2013
    Schlagwort(e): Key words Serotonin ; Inward rectifying potassium channel ; Brain ; PKC ; 5-HT2C receptor ; Heterologous expression ; Xenopus laevis oocytes ; Modulation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract In Xenopus laevis oocytes injected with rat brain poly(A)+ RNA, perfusion with a high-K+ solution (96 mM KCl) generated an inward current (I HK) which was absent in water-injected oocytes. Part of I HK was blocked by low concentrations of Ba2+ (half-maximal inhibitory concentration, IC50: 4.2 ± 0.5 μM). When serotonin (5-HT) was applied to these oocytes a transient inward oscillating Cl− current arising from activation of Ca2+ -dependent Cl− channels, I Cl (Ca), was observed. When this response decayed, a 30% reduction of I HK could be detected. Electrophysiological characterization of the K+ channel down-modulated by 5-HT revealed that it is an inward rectifier. Anti-sense suppression experiments revealed that the 5-HT2C receptor mediates the down-modulatory effect of 5-HT. The nature of the modulatory pathway was investigated by application of phorbol esters and intracellular injection of protein kinase C (PKC) inhibitors, ethylenebis (oxonitrilo)tetraacetate (EGTA) and inositol 1, 4, 5-trisphosphate. The results demonstrate that PKC is responsible for the down-modulatory effect.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF02207270
    Standort Signatur Einschränkungen Verfügbarkeit
    BibTip Andere fanden auch interessant ...
    Artikel (Nationallizenzen)
    Volltext
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