Investigation of K+ channel expression in human peripheral lymphocytes of healthy donors by means of flow cytometry

J. Krjukova, Natalia A Osna, M. Pilmane

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2 Citations (Scopus)

Abstract

Evaluation of different types of K+ channel expression was performed in resting and PHA (phytohemagglutinine)-activated human peripheral lymphocytes (HPL) of healthy donors by means of flow cytometry. In resting peripheral lymphocytes, the application of kaliotoxin (a selective blocker for voltage-dependent K+ (K(V)) channels), K(V) resulted in pronounced depolarization of lymphocyte membrane potential, with further promotion in the presence of thapsigargin (compound discharging Ca(i) from endoplasmic reticulum). In activated HPL, the expression of various types of K+ channels was estimated utilizing cell-cycle analysis data. In contrast to the resting cells, kaliotoxin-induced depolarization of membrane potential in PHA-activated lymphocytes of the G0/G1 phase was not enhanced by thapsigargin and in PHA-activated lymphocytes of the S and G2/M phases we were able to observe repolarization of membrane potential after kaliotoxin-induced depolarization of membrane potential. Substitution of kaliotoxin for charybdotoxin (a non-selective drug blocking both K(V) and K(Ca) channels) abrogated the above effects in PHA-activated lymphocytes. Thus, K(V) channels are active in both resting and activated HPLs and K(Ca) channel expression occurs with cell-cycle progress on PHA-induced activation of peripheral lymphocytes.

Original languageEnglish (US)
Pages (from-to)419-428
Number of pages10
JournalScandinavian Journal of Clinical and Laboratory Investigation
Volume60
Issue number5
DOIs
StatePublished - Jan 1 2000

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Lymphocytes
Flow cytometry
Flow Cytometry
Membrane Potentials
Depolarization
Thapsigargin
Membranes
Cell Cycle
Charybdotoxin
Cells
Cell Cycle Resting Phase
G2 Phase
G1 Phase
Lymphocyte Activation
Endoplasmic Reticulum
Cell Division
Substitution reactions
Chemical activation
kaliotoxin
Pharmaceutical Preparations

Keywords

  • Charybdotoxin
  • Kaliotoxin
  • Membrane potential
  • Thapsigargin

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

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abstract = "Evaluation of different types of K+ channel expression was performed in resting and PHA (phytohemagglutinine)-activated human peripheral lymphocytes (HPL) of healthy donors by means of flow cytometry. In resting peripheral lymphocytes, the application of kaliotoxin (a selective blocker for voltage-dependent K+ (K(V)) channels), K(V) resulted in pronounced depolarization of lymphocyte membrane potential, with further promotion in the presence of thapsigargin (compound discharging Ca(i) from endoplasmic reticulum). In activated HPL, the expression of various types of K+ channels was estimated utilizing cell-cycle analysis data. In contrast to the resting cells, kaliotoxin-induced depolarization of membrane potential in PHA-activated lymphocytes of the G0/G1 phase was not enhanced by thapsigargin and in PHA-activated lymphocytes of the S and G2/M phases we were able to observe repolarization of membrane potential after kaliotoxin-induced depolarization of membrane potential. Substitution of kaliotoxin for charybdotoxin (a non-selective drug blocking both K(V) and K(Ca) channels) abrogated the above effects in PHA-activated lymphocytes. Thus, K(V) channels are active in both resting and activated HPLs and K(Ca) channel expression occurs with cell-cycle progress on PHA-induced activation of peripheral lymphocytes.",
keywords = "Charybdotoxin, Kaliotoxin, Membrane potential, Thapsigargin",
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T1 - Investigation of K+ channel expression in human peripheral lymphocytes of healthy donors by means of flow cytometry

AU - Krjukova, J.

AU - Osna, Natalia A

AU - Pilmane, M.

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N2 - Evaluation of different types of K+ channel expression was performed in resting and PHA (phytohemagglutinine)-activated human peripheral lymphocytes (HPL) of healthy donors by means of flow cytometry. In resting peripheral lymphocytes, the application of kaliotoxin (a selective blocker for voltage-dependent K+ (K(V)) channels), K(V) resulted in pronounced depolarization of lymphocyte membrane potential, with further promotion in the presence of thapsigargin (compound discharging Ca(i) from endoplasmic reticulum). In activated HPL, the expression of various types of K+ channels was estimated utilizing cell-cycle analysis data. In contrast to the resting cells, kaliotoxin-induced depolarization of membrane potential in PHA-activated lymphocytes of the G0/G1 phase was not enhanced by thapsigargin and in PHA-activated lymphocytes of the S and G2/M phases we were able to observe repolarization of membrane potential after kaliotoxin-induced depolarization of membrane potential. Substitution of kaliotoxin for charybdotoxin (a non-selective drug blocking both K(V) and K(Ca) channels) abrogated the above effects in PHA-activated lymphocytes. Thus, K(V) channels are active in both resting and activated HPLs and K(Ca) channel expression occurs with cell-cycle progress on PHA-induced activation of peripheral lymphocytes.

AB - Evaluation of different types of K+ channel expression was performed in resting and PHA (phytohemagglutinine)-activated human peripheral lymphocytes (HPL) of healthy donors by means of flow cytometry. In resting peripheral lymphocytes, the application of kaliotoxin (a selective blocker for voltage-dependent K+ (K(V)) channels), K(V) resulted in pronounced depolarization of lymphocyte membrane potential, with further promotion in the presence of thapsigargin (compound discharging Ca(i) from endoplasmic reticulum). In activated HPL, the expression of various types of K+ channels was estimated utilizing cell-cycle analysis data. In contrast to the resting cells, kaliotoxin-induced depolarization of membrane potential in PHA-activated lymphocytes of the G0/G1 phase was not enhanced by thapsigargin and in PHA-activated lymphocytes of the S and G2/M phases we were able to observe repolarization of membrane potential after kaliotoxin-induced depolarization of membrane potential. Substitution of kaliotoxin for charybdotoxin (a non-selective drug blocking both K(V) and K(Ca) channels) abrogated the above effects in PHA-activated lymphocytes. Thus, K(V) channels are active in both resting and activated HPLs and K(Ca) channel expression occurs with cell-cycle progress on PHA-induced activation of peripheral lymphocytes.

KW - Charybdotoxin

KW - Kaliotoxin

KW - Membrane potential

KW - Thapsigargin

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