Calcium homeostasis and cell death in Sol8 dystrophin-deficient cell line in culture
Related Experiment Video
Updated: Sep 16, 2026
Adult and Embryonic Skeletal Muscle Microexplant Culture and Isolation of Skeletal Muscle Stem Cells
Published on: September 22, 2010
E Marchand1, B Constantin, C Vandebrouck
1Laboratoire des Biomembranes et Signalisation Cellulaire, UMR CNRS/Universi&tacute; de Poitiers, Poitiers, France. eric.marchand@voila.fr
Abstract:
Abnormalities of calcium homeostasis are involved in the process of cell injuries such as Duchenne muscular dystrophy characterized by the absence of the protein dystrophin. But how the absence of dystrophin leads to cytosolic calcium overload is as yet poorly understood. This question has been addressed with skeletal muscle cells from human DMD muscles or mdx mice. Although easier to obtain than human muscles, mdx muscle cells have provided controversial data concerning the resting intracellular calcium level ([Ca2+](i)). This work describes the culture of Sol8 cell line that expresses neither dystrophin nor adhalin, a dystrophin-associated protein. The [Ca2+](i)and intracellular calcium transients induced by different stimuli (acetylcholine, caffeine and high potassium) are normal during the first days of culture. At later stages, calcium homeostasis exhibits drastic alterations with a breaking down of the calcium responses and a large [Ca2+](i)elevation. Concomitantly, Sol8 cells exhibit morphological signs of cell death like cytoplasmic shrinkage and incorporation of propidium iodide. Cell death could be significantly reduced by blocking the activity of calpains, a type of calcium-regulated proteases. These results suggest that Sol8 cell line provides an alternative model of dystrophin-deficient skeletal muscle cells for which a clear disturbance of the calcium homeostasis is observed in culture in association with calpain-dependent cell death. It is shown that transfection with a plasmid cDNA permits the forced expression of dystrophin in Sol8 myotubes as well as a correct sorting of the protein. This approach could be used to explore possible interactions between dystrophin deficiency, calcium homeostasis alteration, and dystrophic cell death.
Insights
Duchenne muscular dystrophy involves calcium problems. A new Sol8 cell model shows that lacking dystrophin leads to calcium overload and cell death, which calpain inhibitors can reduce.
Area of Science:
- Cell Biology
- Neuroscience
- Biochemistry
Background:
- Abnormal calcium homeostasis contributes to cell injury in Duchenne muscular dystrophy (DMD), linked to dystrophin absence.
- The precise mechanism of cytosolic calcium overload due to dystrophin deficiency remains unclear.
- Previous studies using mdx mouse muscle cells yielded conflicting data on resting intracellular calcium levels.
Purpose of the Study:
- To investigate calcium homeostasis in a novel dystrophin-deficient skeletal muscle cell model (Sol8 cell line).
- To characterize calcium handling and cell death mechanisms in Sol8 cells cultured over time.
- To explore the potential of Sol8 cells for studying dystrophin deficiency and related pathologies.
Main Methods:
- Culture of Sol8 cells, which lack both dystrophin and adhalin.
- Measurement of resting intracellular calcium ([Ca2+](i)) and calcium transients induced by acetylcholine, caffeine, and high potassium.
- Assessment of cell death markers (cytoplasmic shrinkage, propidium iodide uptake) and the effect of calpain inhibitors.
- Dystrophin expression via plasmid cDNA transfection in Sol8 myotubes.
Main Results:
- Initially, Sol8 cells exhibited normal calcium homeostasis and responses to stimuli.
- Later in culture, Sol8 cells showed disrupted calcium homeostasis, impaired calcium responses, and elevated [Ca2+](i).
- Dystrophin-deficient Sol8 cells displayed morphological signs of cell death, significantly reduced by calpain inhibition.
Conclusions:
- The Sol8 cell line serves as a valuable model for dystrophin-deficient skeletal muscle, revealing disturbed calcium homeostasis and calpain-dependent cell death in culture.
- This model facilitates the study of the interplay between dystrophin deficiency, calcium dysregulation, and the mechanisms of muscular dystrophy.
- Successful dystrophin re-expression in Sol8 cells offers a platform for further investigation into dystrophin's role in calcium regulation and cell survival.
Related Concept Videos
01:21Satellite Stem Cells and Muscular Dystrophy
01:21Skeleton and Calcium Homeostasis