Temporal and spatial appearance of alpha-dystroglycan in differentiated mouse myoblasts in culture.
The dystrophin-glycoprotein complex plays an important role in muscle function. One of the components of the complex, a 156-kDa cell surface glycoprotein (alpha-dystroglycan) binds to laminin, thereby connecting the basal lamina and muscle cells. We have examined the progressive appearance of alpha-dystroglycan and laminin in muscle cells that differentiate in culture. We find that nondifferentiated cultures of C2C12 myoblasts express low amounts of dystroglycan mRNA and, in contrast, this gene is prominently expressed in differentiated myotubes. Immunofluorescence analysis with a monoclonal antibody against alpha-dystroglycan shows its progressive appearance during myoblast differentiation into myotubes. Immunostaining with a monoclonal antibody against laminin shows that it is not present on the surface of undifferentiated myoblasts. Subsequently, laminin becomes apparent on the surface of differentiated myotubes where it codistributes with immunostained alpha-dystroglycan. Immunoblotting of isolated surface membrane proteins of differentiated myotubes with antibodies against alpha-dystroglycan identifies a broad band of about 140-160 kDa, resembling alpha-dystroglycan from rabbit muscle. The composite results indicate that alpha-dystroglycan and laminin appear and become co-distributed on the surface of cultured C2C12 during the progression of differentiation.
Kostrominova
T Y
TY
Department of BioStructure and Function, School of Dental Medicine, University of Connecticut Health Center, Farmington 06030-3705, USA.
Tanzer
M L
ML
D000818:Animals; D000911:Antibodies, Monoclonal; D002454:Cell Differentiation; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000032:analysis / Q000096:biosynthesis*; D049030:Dystroglycans; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D007150:Immunohistochemistry; D007797:Laminin / Q000096:biosynthesis; D008562:Membrane Glycoproteins / Q000032:analysis / Q000096:biosynthesis*; D051379:Mice; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism; D009132:Muscles; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D011817:Rabbits
D000911:Antibodies, Monoclonal; D003598:Cytoskeletal Proteins; D007797:Laminin; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D049030:Dystroglycans