Rapid mobilization of intracellular Mg2+ by bombesin in Swiss 3T3 cells: mobilization through external Ca(2+)- and tyrosine kinase-dependent mechanisms.
We examined the effects of growth factors on intracellular free Mg2+ concentrations ([Mg2+]i) in single Swiss 3T3 fibroblasts, using microfluorometry of a Mg(2+)-sensitive dye mag-fura-2. We had already noted an increase in [Mg2+]i after exposure to bombesin for 30-60 min [Ishijima, S., Sonoda, T., & Tatibana, M. (1991) Am. J. Physiol. 261 (Cell Physiol. 30), C1074-C1080]. In the present work, we found that bombesin also induced early changes in [Mg2+]i. The [Mg2+]i reached peak values within 15 s in most cells, and the significant rise lasted only for 1-2 min. The extent of the increase varied from cell to cell (0-600 microM above basal). On the average, the [Mg2+]i was increased from basal 0.33 to 0.54 mM. Since the time course was similar to that of [Ca2+]i changes, and the dye mag-fura-2 also binds Ca2+, we evaluated Ca2+ interference with measurement of [Mg2+]i. The contribution of Ca2+ binding would be below 20% of the mag-fura-2 signal. The bombesin-induced [Mg2+]i increase was not dependent on external Mg2+, but the omission of external Ca2+ decreased by 60% the [Mg2+]i increase, and the Ca2+ channel blocker, nicardipine inhibited by 90% the [Mg2+]i response. This inhibition was partially reversed by raising the concentration of external Ca2+. Two structurally distinct tyrosine kinase inhibitors, genistein and lavendustin A, almost completely inhibited the [Mg2+]i response. These results suggest that bombesin rapidly induces Mg2+ mobilization from the intracellular pool, through external Ca(2+)- and tyrosine kinase-dependent mechanisms.
Ishijima
S
S
Department of Biochemistry, Chiba University School of Medicine.
Tatibana
M
M
D016475:3T3 Cells; D000255:Adenosine Triphosphate / Q000378:metabolism; D000468:Alkalies; D000818:Animals; D001839:Bombesin / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D015759:Ionomycin / Q000494:pharmacology; D008274:Magnesium / Q000378:metabolism*; D051379:Mice; D008934:Mitogens / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013997:Time Factors
D000468:Alkalies; D002121:Calcium Channel Blockers; D008934:Mitogens; D015759:Ionomycin; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D011505:Protein-Tyrosine Kinases; D011493:Protein Kinase C; D008274:Magnesium; D001839:Bombesin; D002118:Calcium