Gelsolin sensitivity of microfilaments as a marker for muscle differentiation.
The ability of porcine smooth muscle gelsolin to sever actin filaments was used to study alterations in the organization of F-actin containing structures during skeletal myogenesis. In permeabilized fibroblasts and unfused myoblasts, gelsolin induced complete degradation of the actin cytoskeleton. After fusion of myoblasts to multinucleated myotubes, gelsolin removed a substantial amount of actin, revealing fibers with a sarcomere-like arrangement of gelsolin-insensitive actin. These fibrils were much thinner and had shorter sarcomeres than fully differentiated myofibrils. The proportion of gelsolin-resistant fibrils increased during differentiation, resulting in almost complete inertness of mature myofibrils. Fibrils isolated from adult muscle were also found nearly resistant to gelsolin. Extraction of tropomyosin and myosin in buffer of high ionic strength prior to gelsolin treatment reestablished the susceptibility to the severing protein, both in myotubes and isolated myofibrils. Only small remnants of phalloidin-stainable material were retained. We therefore conclude that during myotube differentiation either an increased interaction of actin with actin-binding proteins (e.g., myosin and tropomyosin), or the assembly of muscle-specific isoforms of these proteins protect the filaments against degradation by actin severing proteins.
Huckriede
A
A
Developmental Biology Unit, University of Bielefeld/Federal Republic of Germany.
Hinssen
H
H
Jockusch
B M
BM
Lazarides
E
E
D008841:Actin Cytoskeleton / Q000187:drug effects*; D000199:Actins / Q000378:metabolism*; D000818:Animals; D002135:Calcium-Binding Proteins / Q000494:pharmacology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D002642:Chick Embryo; D003599:Cytoskeleton / Q000187:drug effects*; D018260:Gelsolin; D008840:Microfilament Proteins / Q000494:pharmacology*; D009132:Muscles / Q000166:cytology* / Q000187:drug effects
D000199:Actins; D002135:Calcium-Binding Proteins; D018260:Gelsolin; D008840:Microfilament Proteins