ACTIVATION OF THE CA2+ MESSAGE SYSTEM BY PARATHYROID-HORMONE IS DEPENDENT ON THE CELL-CYCLE

被引:19
作者
BIZZARRI, C
CIVITELLI, R
机构
[1] WASHINGTON UNIV,MED CTR,JEWISH HOSP,DIV ENDOCRINOL & BONE & MINERAL DIS,ST LOUIS,MO 63110
[2] CONSORZIO BIOLAQ,I-67100 LAQUILA,ITALY
关键词
D O I
10.1210/en.134.1.133
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
To verify whether the heterogeneous intracellular calcium ([Ca2+](i)) responses to PTH observed in the UMR 106-01 osteogenic sarcoma cells are secondary to cell cycle asynchrony or to genotypic differences within the population, we synchronized cell monolayers at the G1/S boundary using a sequential thymidine-aphidicolin block. Video image analysis of fura-2-loaded cells revealed that PTH (10(-7) M) induced transient increases of [Ca2+](i) preferentially in cells in S phase (82% response frequency, n = 63; 286 +/- 33% of baseline, n = 29), whereas cells in G1 phase responded poorly to PTH (10% response frequency, n = 51; 140 +/- 8% of baseline, n = 5). In contrast, cell exposure to 2% fetal calf serum was followed by [Ca2+](i) transients in 83% (n = 42) of cells in G1 phase, but in only 25% (n = 63) of cells in S phase, with similar response amplitude. Hormonal responsiveness was heterogeneous in small clones obtained from single UMR 106-01 cells, with response frequency similar to that observed in nonsynchronized cultures. Pretreatment with either La3+, nifedipine, or pertussis toxin reduced both frequency and amplitude of PTH response in S phase to levels close to G1 phase, whereas there was no significant difference in inositol trisphosphate generated by PTH stimulation in either phase. Therefore, the heterogeneous [Ca2+](i) responses of UMR 106-01 cells to hormonal stimulation is dependent on the phase of the cell cycle, rather than on genotypic heterogeneity. The switch from the G1 to the S phase mode of response is driven by active coupling between the PTH receptor and a Ca2+ channel through a pertussis toxin-sensitive G protein.
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页码:133 / 140
页数:8
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