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Zinc Deficiency Induced in Swiss 3T3 Cells by a Low-Zinc Medium Impairs Calcium Entry and Two Mechanisms of Entry Are Involved

Biological Trace Element Research volume 152, pages 98–104 (2013)Cite this article

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.

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Abbreviations

BSA:

Bovine serum albumin

DAG:

Diacylglycerol

DTPA:

Diethylenetriaminepentaacetate

EGF:

Epidermal growth factor

GF:

Growth factors

IGF-I:

Insulin-like growth factor-I

IP3:

Inositol 1,4,5-trisphosphate

OMEM:

Low-zinc, low-calcium medium

PDGF:

Platelet-derived growth factor

ROCE:

Receptor-operated calcium entry

SOCE:

Store-operated calcium entry

STIM1:

Stromal interaction molecule 1, a calcium sensing protein

TG:

Thapsigargin

TRPC:

Transient receptor potential canonical

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Authors and Affiliations

  1. Department of Biochemistry, University of Missouri, 256 Stringer Wing, Eckles Hall, Columbia, MO, 65211, USA

    Boyd L. O’Dell

  2. Department of Animal Science, University of Missouri, Columbia, MO, 65211, USA

    Jimmy D. Browning

Authors
  1. Boyd L. O’Dell
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  2. Jimmy D. Browning
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Corresponding author

Correspondence to Boyd L. O’Dell.

Additional information

This study was supported by the University of Missouri Food for the 21st Century Program.

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O’Dell, B.L., Browning, J.D. Zinc Deficiency Induced in Swiss 3T3 Cells by a Low-Zinc Medium Impairs Calcium Entry and Two Mechanisms of Entry Are Involved. Biol Trace Elem Res 152, 98–104 (2013). https://doi.org/10.1007/s12011-012-9591-6

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  • DOI: https://doi.org/10.1007/s12011-012-9591-6

Keywords

  • Zinc deficiency
  • Calcium entry
  • Cell proliferation
  • Zinc–calcium interaction