Immunological stimulation of single rat basophilic leukemia RBL-2H3 cells co-activates Ca(2+)-entry and K(+)-channels.
Cell Calcium
; 17(1): 71-83, 1995 Jan.
Article
in En
| MEDLINE
| ID: mdl-7553783
ABSTRACT
The relationship between type 1 Fc epsilon-receptor (Fc epsilon RI) mediated cell stimulation, Ca(2+)-signals and membrane currents was studied in rat mucosal mast cells, subline RBL-2H3 by combining patch-clamp, Fura-2, 45Ca(2+)-uptake and secretory response measurements. Cells were stimulated by Fc epsilon RI clustering either with IgE and antigen or by an IgE specific monoclonal antibody. Both stimuli induced a biphasic increase in the free intracellular Ca(2+)-concentration ([Ca2+]i). Fc epsilon RI clustering in Ca(2+)-free solution induces a transient increase in [Ca2+]i reflecting Ca2+ release from the Ins(1,4,5)P3 sensitive stores. Mn2+ applied to a nominally Ca(2+)-free solution, causes quenching of the Fura-2 emission during Fc epsilon RI clustering, indicating activation of a transmembrane pathway for the entry of extracellular calcium ions. Whole-cell current-voltage relationship of resting cells showed strong inward rectification. The inward current component at a potential of -100 mV is increased by 23 +/- 11% (n = 14) upon Fc epsilon RI clustering, whereas the outward component at +50 mV was enhanced by 45 +/- 6%. The Fc epsilon RI activated current was identified as due to K+ ions, because it reversed close to the K(+)-equilibrium potential, was blocked by Ba2+ or Cs+ containing or K(+)-free bath solutions. It was also inhibited by TEA and quinidine, while DIDS had no effect. Moreover, an inwardly rectifying K(+)-channel with a conductance of 28 pS was recorded in single channel measurements. The open probability of this channel increased by 39 +/- 16% (n = 8) upon Fc epsilon RI clustering. Superfusion of the cells with nominally K(+)-free solution also significantly inhibited both the Fc epsilon RI mediated 45Ca2+ uptake and the secretory response of the cells. We conclude that activation of K(+)-channels upon Fc epsilon RI clustering is functionally involved in the control and the maintenance of the secretory response of RBL-2H3 mast cells.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Potassium Channels
/
Calcium
/
Receptors, IgE
Type of study:
Prognostic_studies
Limits:
Animals
Language:
En
Journal:
Cell Calcium
Year:
1995
Document type:
Article