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

• Published in: Cell and tissue research 1991-07, Vol.265 (1), p.95-103
• Main Authors: BURSZTAJN, S, SCHNEIDER, L. W, YUN-JIN JONG, BERMAN, S. A
• Format: Article
• Language: English
• Subjects: Acetylcholinesterase - metabolism; Animals; Autoradiography; Biological and medical sciences; Calcimycin - pharmacology; Calcium - pharmacology; Cells, Cultured; Chick Embryo; Electrophoresis, Polyacrylamide Gel; Extracellular Matrix - metabolism; Extracellular Matrix - ultrastructure; Fundamental and applied biological sciences. Psychology; Microscopy, Electron, Scanning; Muscles - cytology; Muscles - embryology; Muscles - metabolism; Secretin - metabolism; Striated muscle. Tendons; Vertebrates: osteoarticular system, musculoskeletal system
• ISSN: 0302-766X; 1432-0878
• DOI: 10.1007/BF00318143

Calcium (Ca2+) and calcium-transporting ionophores stimulate protein secretion in many cellular systems. We demonstrate here than 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 65,000 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.