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Ca2(+)‐activated K+ channels in human B lymphocytes and rat thymocytes.
Author(s) -
MahautSmith M P,
Schlichter L C
Publication year - 1989
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1113/jphysiol.1989.sp017712
Subject(s) - pipette , ionomycin , conductance , biophysics , chemistry , membrane potential , patch clamp , potassium channel , calcium activated potassium channel , intracellular , biochemistry , biology , receptor , mathematics , combinatorics
1. Previous evidence for the existence of Ca2(+)‐activated K+ channels in lymphocytes comes from measurements using voltage‐sensitive dyes and from tracer flux studies. We have now directly measured these channels in human tonsillar B lymphocytes and rat thymocytes in single‐channel recordings from cell‐attached and excised patches. 2. In cell‐attached recordings, intracellular Ca2+ was raised by either ionomycin or replacement of external Ca2+ following incubation in Ca2(+)‐free medium. Indo‐1 measurements during the Ca2(+)‐replacement technique showed that [Ca2+]i rose from approximately 90 to 260 nM. Both techniques activated two channels of approximately 25 and 8 pS (slope conductance at 0 mV applied, with 140 mM‐K+ in the pipette). Over 90% of patches displayed this activity, indicating a high density of these channels in the membrane. 3. Both channels reversed near the K+ equilibrium potential with either KCl or potassium aspartate in the pipette, when the cells were bathed in normal or high‐K+ saline. Therefore, these channels are selective for K+. 4. The larger channel was studied in more detail. It displayed inward rectification in symmetrical K+ solutions. The open‐channel probability was weakly dependent on membrane potential. 5. Ca2(+)‐dependent K+ channels were also recorded from excised, inside‐out membrane patches. The threshold for activation was 200‐300 nM [Ca2+i]. 6. Patch excision altered some characteristics of IK(Ca). Channels were activated in fewer than 50% of patches and the main conductance level was approximately 34 pS (at ‐80 mV). The duration of single‐channel events was shorter than in cell‐attached patches; kinetic analysis suggested that this was due to the loss of an open state in excised patches. 7. We conclude that B and T lymphocytes have K(+)‐selective channels which are activated by internal [Ca2+] in the physiological range and which will influence the membrane potential during cell activation.