Differential intracellular distribution and activities of mu- and m-calpains during the differentiation of human myogenic cells in culture.
Calpains are intracellular calcium-dependent cystein proteases active at neutral pH. There have been found in human adult myogenic cells (i.e. satellite cells) 2 forms of calpains requiring either micromolar Ca2+: mu-calpain, or millimolar Ca2+: m-calpain. Calpains could be involved in both intracellular proteolysis and cytoskeleton reorganization required for myogenic cell fusion. We showed significant differences in calpains distribution during differentiation of myogenic cells. Using mono- and polyclonal antibodies against both types of calpains, we localized mu-calpain and m-calpain in cultured human satellite cells. mu-calpain was detected in the nuclei of myoblasts and in the cytoplasm of myotubes. m-calpain was only present in the cytoplasm, and was concentrated near the nuclear envelope. Biochemical assays for calpain activities showed that the amounts of these proteinases were modulated during cell growth and differentiation. m-calpain activity was high at the proliferation phase (day 4 of culture) and reached a maximum with the beginning of fusion (day 8) and decreased slightly when the number of myotubes increased (day 12). This activity profile suggests that m-calpain could play a role in the initiation of fusion of satellite cells. The activity of mu-calpain increased regularly with cell growth, the maximum being reached when the cells differentiate, i.e. when its intracellular localization shifted from the nucleus to the cytoplasm. We conclude that the activity and the intracellular localization of the 2 forms of calpains differ with the state of differentiation of myogenic cells.
Moraczewski
J
J
Faculty of Biology, University of Warsaw, Poland.
Piekarska
E
E
Bonavaud
S
S
Wosinska
K
K
Chazaud
B
B
Barlovatz-Meimon
G
G
D002154:Calpain / Q000032:analysis* / Q000378:metabolism*; D002454:Cell Differentiation; D002455:Cell Division; D002467:Cell Nucleus / Q000737:chemistry; D002478:Cells, Cultured; D003593:Cytoplasm / Q000737:chemistry*; D006801:Humans; D066298:In Vitro Techniques; D018482:Muscle, Skeletal / Q000166:cytology*
D002154:Calpain