Andrzej Teisseyre

The voltage- and time-dependent blocking effect of trifluoperazine on T lymphocyte Kv1.3 channels

Phenothiazines are well-known calmodulin inhibitors that interact with many receptors and channels including a variety of potassium channels. In this study, we report a blocking effect of trifluoperazine (TFP) on voltage-gated Kv1.3 channels expressed in human T lymphocytes. Application of TFP in the concentration range from 1 to 20 microM reduced the current amplitude to about a half of the control value. The currents were blocked to less than 0.05 of the control value at 50 microM TFP concentration. The blocking effect was accompanied by a substantial increase in the current inactivation rate, whereas the activation rate and the steady-state activation and inactivation were not changed significantly. The blocking effect of TFP was voltage dependent being most potent at +60mV and least potent at -20mV. The blocking effect of TFP on the currents and the recovery from block was time dependent. Other calmodulin antagonists: tamoxifen (TMX) and thioridazine also inhibited the channels at micromolar concentrations. The effects exerted by TMX and thioridazine resembled the inhibitory effect of TFP. The blocking effect of thioridazine was time dependent and appeared to be more potent that the inhibition by TFP and TMX. TFP, TMX and thioridazine inhibited the activity of Kv1.3 channels only when applied extracellularly. The inhibitory effect of all the compounds was reversible. The possible physiological significance of the current inhibition is discussed.

The influence of protons and zinc ions on the steady-state inactivation of Kv1.3 potassium channels

Using the whole-cell patch-clamp technique, we investigated the influence of extracellular pH and zinc ions (Zn2+) on the steady-state inactivation of Kv1.3 channels expressed in human lymphocytes. The obtained data showed that lowering the extracellular pH from 7.35 to 6.8 shifted the inactivation midpoint (Vi) by 17.4 ± 1.12 mV (n = 6) towards positive membrane potentials. This shift was statistically significant (p < 0.05). Applying 100 μM Zn2+ at pH 6.8 further shifted the Vi value by 16.55 ± 1.80 mV (n = 6) towards positive membrane potentials. This shift was also statistically significant (p < 0.05). The total shift of the Vi by protons and Zn2+ was 33.95 ± 1.90 mV (n = 6), which was significantly higher (p < 0.05) than the shift caused by Zn2+ alone. The Zn2+-induced shift of the Vi at pH 6.8 was almost identical to the shift at pH = 7.35. Thus, the proton-and Zn2+-induced shifts of the Vi value were additive. The steady-state inactivation curves as a function of membrane voltage were compared with the functions of the steady-state activation. The total shift of the steady-state inactivation was almost identical to the total shift of the steady-state activation (32.01 ± 2.10 mV, n = 10). As a result, the “windows” of membrane potentials in which the channels can be active under physiological conditions were also markedly shifted towards positive membrane potentials. The values of membrane voltage and the normalised chord conductance corresponding to the points of intersection of the curves of steady-state activation and inactivation were also calculated. The possible physiological significance of the observed modulatory effects is discussed herein.

The influence of 8-prenylnaringenin on the activity of voltage-gated kv1.3 potassium channels in human jurkat t cells

Using the whole-cell patch-clamp technique, we investigated the influence of 8-prenylnaringenin on the activity of the voltage-gated Kv1.3 potassium channels in the human leukemic T lymphocyte cell line Jurkat. 8-prenylnaringenin is a potent plant-derived phytoestrogen that has been found to inhibit cancer cell proliferation. The results show that it inhibited the Kv1.3 channels in a concentration-dependent manner. Complete inhibition occurred at concentrations higher than 10 μM. The inhibitory effect of 8-prenylnaringenin was reversible. It was accompanied by a significant acceleration of channel inactivation without any pronounced change in the activation rate. Of the naringenin derivatives tested to date, 8-prenylnaringenin is the most potent inhibitor of the Kv1.3 channels. The potency of the inhibition may be due to the presence of a prenyl group in the molecule of this flavonoid. The inhibition of the Kv1.3 channels might be involved in the antiproliferative and pro-apoptotic effects of 8-prenylnaringenin that have been observed in cancer cell lines expressing these channels.

Patch-clamp study on T-lymphocyte potassium conductance in patients with chronic renal failure.

Using the patch-clamp technique, we studied the differences in whole-cell potassium conductance (g(K)+) in T lymphocytes (TL) from three groups of patients suffering from renal failure: not dialyzed patients, dialyzed patients, and dialyzed patients treated with human recombinant erythropoietin (rHuEPO). The differences in g(K+) values in the group of not dialyzed patients in comparison with controls was not significant (p > 0.05). In the group of dialyzed patients, after roughly 6 years of the hemodialysis therapy, the g(K+) value was significantly higher than in controls. In dialyzed patients treated with rHuEPO, g(K+) value was significantly lower in comparison with control. Moreover, in dialyzed patients treated with rHuEPO, the time duration of dialysis therapy did not significantly affect the TL whole-cell conductance. We conclude that the g(K+) is changed in TL in renal failure patients and that the time duration of hemodialysis therapy as well as the use of rHuEPO affect the g(K+) value. Possible mechanisms underlying the observed changes in g(K+) values, as well as medical implications of obtained results are discussed.