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Calcium and ionophore A23187 stimulates deposition of extracellular matrix and acetylcholinesterase release in cultured myotubes

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Summary

Calcium (Ca2+) and calcium-transporting ionophores stimulate protein secretion in many cellular systems. We demonstrate here that increases in intracellular calcium concentration induce a time- and concentration-dependent deposition of extracellular matrix and an increase in acetylcholinesterase secretion. Scanning and transmission electron-microscopy revealed that treatment with the calcium ionophore A23187, or high extracellular Ca2+ levels (5 mM to 15 mM) produce significant deposits of extracellular matrix around the myotubes, as well as a marked increase in the acetylcholinesterase reaction-product. Blocking muscle contraction was not necessary for the induction of AChE secretory activity. Sucrose density-gradients of media conditioned by muscle cells revealed 3 separate acetylcholinesterase molecular forms. However, incubation with A23187 increased only the 4.5 S and the 7.2 S molecular forms, whereas the 12.0 S form showed no significant differences from controls. Polyacrylamide gel electrophoresis, and autoradiography using [3H]diisopropyl fluorophosphate revealed a broad band at 65000 daltons. This band was broader than for controls when medium was obtained from A23187-treated cells. Our results show that increasing intracellular Ca2+ concentration induces marked deposition of extracellular matrix and increased acetylcholinesterase secretion, with an apparent selectivity for the monomeric and dimeric acetylcholinesterase molecular forms.

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Authors and Affiliations

  1. Department of Psychiatry, Harvard Medical School, Mailman Research Center, 02178, Belmont, MA, USA

    Sherry Bursztajn

  2. Department of Neurology, Baylor College of Medicine, 77030, Houston, TX, USA

    Larry W. Schneider & Yun-Jin Jong

  3. Spinal Cord Injury Service, Harvard Medical School, West Roxbury VA Hospital, 02132, Boston, MA, USA

    Stephen A. Berman

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  1. Sherry Bursztajn
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  2. Larry W. Schneider
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  3. Yun-Jin Jong
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  4. Stephen A. Berman
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Bursztajn, S., Schneider, L.W., Jong, YJ. et al. Calcium and ionophore A23187 stimulates deposition of extracellular matrix and acetylcholinesterase release in cultured myotubes. Cell Tissue Res 265, 95–103 (1991). https://doi.org/10.1007/BF00318143

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