Remodelling of the nuclear periphery during muscle cell differentiation in vitro.
We have examined the composition and ultrastructure of the nuclear periphery during in vitro myogenesis of the rat myoblast cell line, L6E9. Immunofluorescence labelling and immunoblotting showed that lamins A/C and B were all present in undifferentiated cells, but that they increased significantly before extensive cell fusion had occurred, with lamins A/C increasing proportionately more. Electron microscopic observations were consistent with these results, showing an increase in the prominence of the lamina during differentiation. On the other hand, immunofluorescence labelling suggested that the P1 antigen began to disappear from the nuclear periphery as the cells were fusing, after the increase in lamin quantity, and was no longer detectable in multinucleated cells. Unexpectedly, however, P1 was readily detected in isolated nuclei, whether prepared from myoblast or differentiated cultures, as well as in both myoblast and myotube nuclear matrices. It appears probable, therefore, that the fading of P1 labelling is due to masking of the epitope by a soluble factor recruited to the nuclear periphery as cell differentiate. These data, together with evidence that the genome is substantially rearranged during L6E9 myogenesis [Chaly and Munro, 1996], suggest that L6E9 cells are a useful model system in which to study the interrelationship of nuclear envelope organization, chromatin spatial order, and nuclear function.
Chaly
N
N
Department of Biology, Carleton University, Ottawa, Ontario, Canada.
Munro
S B
SB
Swallow
M A
MA
D000818:Animals; D002453:Cell Cycle; D002454:Cell Differentiation; D002460:Cell Line; D002467:Cell Nucleus; D002850:Chromatography, Gel; D019084:Fluorescent Antibody Technique, Indirect; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D008954:Models, Biological; D009132:Muscles / Q000648:ultrastructure*; D015530:Nuclear Matrix / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats