title: The absence of MyoD in regenerating skeletal muscle affects the expression pattern of basement membrane, interstitial matrix and integrin molecules that is consistent with delayed myotube formation.
abstract: MyoD is a member of a skeletal muscle specific family of transcription factors which directs the events of myogenesis during development and regeneration. Muscle cells that lack MyoD show delayed fusion in vivo and in vitro and defects have been observed in vitro in the attachment of MyoD(-/-) myoblasts to complex substrates such as Matrigel. Since interactions with the extracellular matrix (ECM) are important during myoblast fusion (i. e. myotube formation), it was hypothesised that expression of ECM molecules or their receptors may be altered in MyoD(-/-) muscle. The production of basement membrane molecules such as collagen type IV and several laminins, the interstitial molecules fibronectin and tenascin-C, and the cell surface molecules integrin alpha5 and alpha6 were quantitated in vitro using ELISA on cultured cells from MyoD(-/-) and wild type mice. Differences were observed in the production of fibronectin, tenascin-C, collagen type IV, laminin-1 and integrin alpha5 between control and MyoD(-/-) myotubes in vitro. This corresponded with delayed fusion of myoblasts in MyoD(-/-) cultures. On the basis of these findings with respect to matrix expression in vitro, fluorescent immunohistochemistry was carried out on adult whole muscle autografts to examine whether the expression of these molecules, as well as integrin alpha7, was altered in the complex in vivo environment. Some minor differences in expression patterns were observed in MyoD(-/-) as compared to normal BALB/c autografts. The overall expression of matrix components was consistent with the delayed onset of myotube formation. These results suggest that the delay in myotube formation in MyoD(-/-) muscle is not a direct result of altered expression of the matrix molecules collagen type IV, laminins, fibronectin, tenascin-C, and integrins alpha5, alpha6 or alpha7.
authors.affiliation: Department of Anatomy and Human Biology, The University of Western Australia, Crawley, Australia.
authors.forename: J
authors.initials: J
authors.lastname: Huijbregts
authors.forename: J D
authors.initials: JD
authors.lastname: White
authors.forename: M D
authors.initials: MD
authors.lastname: Grounds
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D015703:Antigens, CD / Q000235:genetics; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005109:Extracellular Matrix / Q000502:physiology; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D039482:Integrin alpha5; D007797:Laminin / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D018482:Muscle, Skeletal / Q000166:cytology / Q000502:physiology* / Q000637:transplantation; D017570:MyoD Protein / Q000235:genetics* / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D012038:Regeneration / Q000502:physiology*; D019063:Tenascin / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D024141:Collagen Type IV; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D039482:Integrin alpha5; D007797:Laminin; D017570:MyoD Protein; D019063:Tenascin