title: Zinc deficiency induced in Swiss 3T3 cells by a low-zinc medium impairs calcium entry and two mechanisms of entry are involved.
abstract: Zinc deficiency in 3T3 cells induced by the use of diethylenetriaminepentaacetate (DTPA) has been shown to impair calcium entry associated with failure of proliferation when the cells are stimulated with polypeptide growth factors (GF). These functions of zinc have been evaluated here in the same clone of cells by simple depletion using a low-zinc medium (0.05 μmol/L zinc) without chelator. Confluent cells were maintained for 1 day in the low-zinc medium without GF, then loaded with Fluo-4, and stimulated with GF. Calcium entry was measured by the increase in sustained fluorescence. It was preceded by the release of stored calcium as observed in the previous study using DTPA. Zinc deprivation decreased calcium entry when calcium was added at 0 or 0.05 mmol/L but not when 0.1 mmol/L or higher. Cell proliferation reflected similar effects of zinc and calcium concentrations. In a newly acquired clone of 3T3 cells, GF did not induce internal calcium release but thapsigargin (TG) did. When added in a low-calcium medium, both agonists stimulated calcium entry when external calcium was added, suggesting that two different mechanisms of entry were impaired by zinc deficiency. Zinc deficiency produced by DTPA in the newer clones gave similar results, decreasing calcium entry induced by both agonists. The effects of GF and TG were not additive. The results confirm the earlier observation that zinc deficiency impairs calcium entry into 3T3 cells when stimulated by GF and show that the cells can take up calcium by either store-operated or receptor-operated mechanisms.
authors.affiliation: Department of Biochemistry, University of Missouri, 256 Stringer Wing, Eckles Hall, Columbia, MO 65211, USA. odellb@missouri.edu
authors.forename: Boyd L
authors.initials: BL
authors.lastname: O'Dell
authors.forename: Jimmy D
authors.initials: JD
authors.lastname: Browning
mesh_terms: D016475:3T3 Cells; D000814:Aniline Compounds / Q000378:metabolism; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002614:Chelating Agents / Q000494:pharmacology; D003470:Culture Media / Q000378:metabolism* / Q000494:pharmacology; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D051379:Mice; D004369:Pentetic Acid / Q000494:pharmacology; D019284:Thapsigargin / Q000494:pharmacology; D014966:Xanthenes / Q000378:metabolism; D015032:Zinc / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D000814:Aniline Compounds; D002614:Chelating Agents; D003470:Culture Media; D004791:Enzyme Inhibitors; C409648:Fluo 4; D005456:Fluorescent Dyes; D036341:Intercellular Signaling Peptides and Proteins; D014966:Xanthenes; D019284:Thapsigargin; D004369:Pentetic Acid; D000252:Calcium-Transporting ATPases; D015032:Zinc; D002118:Calcium