Subcloning the RBL-2H3 mucosal mast cell line reduces Ca2+ response heterogeneity at the single-cell level.
Ca2+ imaging experiments have revealed that for a wide variety of cell types, including RBL-2H3 mucosal mast cells, there are considerable cell-to-cell differences of the Ca2+ responses of individual cells. This heterogeneity is evident in both the shape and latency of the responses. Mast cells within a single microscopic field of view, which have experienced identical culture conditions and experimental preparation, display a wide variety of responses upon antigen stimulation. We have subcloned the RBL-2H3 mucosal mast cell line to test the hypothesis that genetic heterogeneity within the population is the cause of the Ca2+ response heterogeneity. We found that cell-to-cell variability was significantly reduced in four of five clonal lines. The response heterogeneity remaining within the clones was not an experimental artifact caused by differences in the amount of fura-2 loaded by individual cells. Factors other than genetic heterogeneity must partly account for Ca2+ response heterogeneity. It is possible that the complex shapes and variability of the Ca2+ responses are reflections of the fact that there are multiple factors underlying the Ca2-response to antigen stimulation. Small differences from cell to cell in one or more of these factors could be a cause of the remaining Ca2+ response heterogeneity.
Kuchtey
J
J
Department of Pharmacology, Cornell University, Ithaca, New York 14853, USA.
Fewtrell
C
C
D000818:Animals; D000941:Antigens; D002118:Calcium / Q000378:metabolism*; D002999:Clone Cells / Q000378:metabolism*; D005456:Fluorescent Dyes; D016257:Fura-2; D018740:Genetic Heterogeneity; D007091:Image Processing, Computer-Assisted; D015471:Leukemia, Basophilic, Acute; D008407:Mast Cells / Q000166:cytology / Q000378:metabolism*; D009092:Mucous Membrane; D051381:Rats; D014407:Tumor Cells, Cultured; D005719:gamma-Globulins
D000941:Antigens; D005456:Fluorescent Dyes; C030730:dinitrophenyl gamma globulin; D005719:gamma-Globulins; D002118:Calcium; D016257:Fura-2