Measurement of intracellular Ca2+ in BC3H-1 muscle cells with Fura-2: Relationship to acetylcholine receptor synthesis

J. R. Berlin, M. A. Wozniak, M. B. Cannell, R. J. Bloch, W. J. Lederer

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7 Scopus citations

Abstract

Synthesis of acetylcholine receptors (AChR) can be affected by calcium, but the role played by this cation is controversial. The effect of changes in extracellular calcium, [Ca2+]o, on AChR synthesis was examined in a cultured mouse muscle cell line, BC3H-1. Reduction of [Ca2+]o for long periods (∼22 h) leads to a decrease in total surface AChR levels, a finding that is consistent with inhibition of AChR synthesis. A half-maximal reduction in surface AChR levels is observed when [Ca2+]o is decreased from 1.8 to ∼ 50 μ M. Under these conditions, however, total protein synthesis is also largely inhibited, suggesting that the effect of [Ca2+]o on AChR synthesis may be relatively non-specific. Increasing [Ca2+]i by adding the Ca2+ ionophore, A23187 (in the presence of 1.8 mM [Ca2+]o) also gives similar and significant reductions of both AChR and protein synthesis. Since the time course of changes in intracellular calcium ([Ca2+]i) produced by these manoeuvres is unknown, we examined the effects of briefer (1-6 h) reductions in [Ca2+]o and achieved a more specific reduction in AChR synthesis. A direct measurement of the changes in [Ca2+]i resulting from changes in [Ca2+]o was made using the fluorescent indicator Fura-2 and video fluorescence microscopy. Our results show that in BC3H-1 muscle cells the resting intracellular calcium decreases reversibly over 20 min when [Ca2+]o is decreased. We suggest that a reduction of [Ca2+]i produced by the lower [Ca2+]o underlies the reduction in AChR synthesis observed in these experiments.

Original languageEnglish (US)
Pages (from-to)371-384
Number of pages14
JournalCell Calcium
Volume11
Issue number5
DOIs
StatePublished - May 1990
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Physiology
  • Molecular Biology
  • Cell Biology

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