109740072000101020240922
0021-952515052000Sep04The Journal of cell biologyJ Cell BiolThe dystrophin complex forms a mechanically strong link between the sarcolemma and costameric actin.120912141209-14The absence of dystrophin complex leads to disorganization of the force-transmitting costameric cytoskeleton and disruption of sarcolemmal membrane integrity in skeletal muscle. However, it has not been determined whether the dystrophin complex can form a mechanically strong bond with any costameric protein. We performed confocal immunofluorescence analysis of isolated sarcolemma that were mechanically peeled from skeletal fibers of mouse hindlimb muscle. A population of gamma-actin filaments was stably associated with sarcolemma isolated from normal muscle and displayed a costameric pattern that precisely overlapped with dystrophin. However, costameric actin was absent from all sarcolemma isolated from dystrophin-deficient mdx mouse muscle even though it was localized to costameres in situ. Vinculin, alpha-actinin, beta-dystroglycan and utrophin were all retained on mdx sarcolemma, indicating that the loss of costameric actin was not due to generalized membrane instability. Our data demonstrate that the dystrophin complex forms a mechanically strong link between the sarcolemma and the costameric cytoskeleton through interaction with gamma-actin filaments. Destabilization of costameric actin filaments may also be an important precursor to the costamere disarray observed in dystrophin-deficient muscle. Finally, these methods will be broadly useful in assessing the mechanical integrity of the membrane cytoskeleton in dystrophic animal models lacking other costameric proteins.RybakovaI NINDepartment of Physiology, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA.PatelJ RJRErvastiJ MJMengR01 AR042423ARNIAMS NIH HHSUnited StatesR56 AR042423ARNIAMS NIH HHSUnited StatesAR01985ARNIAMS NIH HHSUnited StatesAR42423ARNIAMS NIH HHSUnited StatesJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.
United StatesJ Cell Biol03753560021-95250Actins0DystrophinIMActinsanalysismetabolismultrastructureAnimalsDystrophinmetabolismultrastructureMiceMice, Inbred C57BLMice, Inbred mdxModels, BiologicalMuscle Fibers, SkeletalphysiologyultrastructureMuscle, SkeletalphysiologyultrastructureSarcolemmametabolismultrastructure20009711020001014111200097110200134ppublish10974007PMC217526310.1083/jcb.150.5.1209Amann K.J., Guo W.X.A., Ervasti J.M. Utrophin lacks the rod domain actin binding activity of dystrophin. J. Biol. Chem. 1999;274:35375–35380.10585405Amann K.J., Renley B.A., Ervasti J.M. A cluster of basic repeats in the dystrophin rod domain binds F-actin through an electrostatic interaction. J. Biol. Chem. 1998;273:28419–28423.9774469Andra K., Lassman H., Bittner R., Shorny S., Fassler R., Propst F., Wiche G. Targeted inactivation of plectin reveals essential function in maintaining the integrity of skin, muscle, and heart cytoarchitecture. Genes Dev. 1997;11:3143–3156.PMC3167469389647Campbell K.P. Three muscular dystrophiesLoss of cytoskeleton-extracellular matrix linkage. Cell. 1995;80:675–679.7889563Chamberlain J.S., Corrado K., Rafael J.A., Cox G.A., Hauser M., Lumeng C. Interactions between dystrophin and the sarcolemma membrane. In: Froehner S.C., Bennett V., editors. Cytoskeletal Regulation of Membrane Function. The Rockefeller University Press; New York: 1997. pp. 19–29.9210217Cote P.D., Moukhles H., M.Lindenbaum, Carbonetto S. Chimaeric mice deficient in dystroglycans develop muscular dystrophy and have disrupted myoneural synapses. Nat. Genet. 1999;23:338–342.10610181Craig S.W., Pardo J.V. Gamma actin, spectrin, and intermediate filament proteins colocalize with vinculin at costameres, myofibril-to-sarcolemma attachment sites. Cell Motility. 1983;3:449–462.6420066Dalpe G., Mathieu M., Comtois A., Zhu E., Wasiak S., De Repetigny Y., Leclerc N., Kothary R. Dystonin-deficient mice exhibit an intrinsic muscle weakness and an instability of skeletal muscle cytoarchitecture. Dev. Biol. 1999;210:367–380.10357897Danowski B.A., Imanaka-Yoshida K., Sanger J.M., Sanger J.W. Costameres are sites of force transmission to the substratum in adult rat cardiomyocytes. J. Cell Biol. 1992;118:1411–1420.PMC22896041522115Duclos F., Straub V., Moore S.A., Venzke D.P., Hrstka R.F., Crosbie R.H., Durbeej M., Lebakken C.S., Ettinger A.J., Van der Meulen J. Progressive muscular dystrophy in α-sarcoglycan-deficient mice. J. Cell Biol. 1998;142:1461–1471.PMC21417739744877Ehmer S., Herrmann R., Bittner R., Voit T. Spatial distribution of β-spectrin in normal and dystrophic human skeletal muscle. Acta Neuropathol. (Berl.) 1997;94:240–246.9292693Ervasti J.M., Campbell K.P. Membrane organization of the dystrophin-glycoprotein complex. Cell. 1991;66:1121–1131.1913804Ervasti J.M., Campbell K.P. A role for the dystrophin-glycoprotein complex as a transmembrane linker between laminin and actin. J. Cell Biol. 1993;122:809–823.PMC21195878349731Ervasti J.M., Ohlendieck K., Kahl S.D., Gaver M.G., Campbell K.P. Deficiency of a glycoprotein component of the dystrophin complex in dystrophic muscle. Nature. 1990;345:315–319.2188135Howard P.L., Klamut H.J., Ray P.N. Identification of a novel actin binding site within the Dp71 dystrophin isoform. Febs Letters. 1998;441:337–341.9883911Iwata Y., Pan Y., Yoshida T., Hanada H., Shigekawa M. α1-syntrophin has distinct binding sites for actin and calmodulin. Febs Letters. 1998;423:173–177.9512352Kameya S., Miyagoe Y., Nonaka I., Ikemoto T., Endo M., Hanaoka K., Nabeshima Y., Takeda S. α1-syntrophin gene disruption results in the absence of neuronal-type nitric-oxide synthase at the sarcolemma but does not induce muscle degeneration. J. Biol. Chem. 1999;274:2193–2200.9890982Lessard J.L. Two monoclonal antibodies to actinOne muscle selective and one generally reactive. Cell Motil. Cytoskel. 1988;10:349–362.2460261Milner D.J., Weitzer G., Tran D., Bradley A., Capetanaki Y. Disruption of muscle architecture and myocardial degeneration in mice lacking desmin. J. Cell Biol. 1996;134:1255–1270.PMC21209728794866Minetti C., Tanji K., Bonilla E. Immunologic study of vinculin in Duchenne muscular dystrophy. Neurology. 1992;42:1751–1754.1513465Minetti C., Tanji K., Rippa P.G., Morreale G., Cordone G., Bonilla E. Abnormalities in the expression of β-spectrin in Duchenne muscular dystrophy. Neurology. 1994;44:1149–1153.8208414Ohlendieck K., Campbell K.P. Dystrophin-associated proteins are greatly reduced in skeletal muscle from mdx mice. J. Cell Biol. 1991;115:1685–1694.PMC22891971757468Ohlendieck K., Ervasti J.M., Matsumura K., Kahl S.D., Leveille C.J., Campbell K.P. Dystrophin-related protein is localized to neuromuscular junctions of adult skeletal muscle. Neuron. 1991;7:499–508.1654951Otey C.A., Bulinski J.C. Immunolocalization of muscle and nonmuscle isoforms of actin in myogenic cells and adult skeletal muscle. Cell Motil. Cytoskel. 1988;9:337–348.3292062Pardo J.V., D'Angelo Siliciano J., Craig S.W. A vinculin-containing cortical lattice in skeletal muscleTransverse lattice elements (“costameres”) mark sites of attachment between myofibrils and sarcolemma Proc. Natl. Acad. Sci. USA. 80 1983. 1008 1012aPMC3935176405378Pardo J.V., Pittenger M.F., Craig S.W. Subcellular sorting of isoactinsselective association of γ actin with skeletal muscle mitochondria Cell. 32 1983. 1093 1103b6340835Petrof B.J., Shrager J.B., Stedman H.H., Kelly A.M., Sweeney H.L. Dystrophin protects the sarcolemma from stresses developed during muscle contraction. Proc. Natl. Acad. Sci. USA. 1993;90:3710–3714.PMC463718475120Porter G.A., Dmytrenko G.M., Winkelmann J.C., Bloch R.J. Dystrophin colocalizes with β-spectrin in distinct subsarcolemmal domains in mammalian skeletal muscle. J. Cell Biol. 1992;117:997–1005.PMC22894901577872Rybakova I.N., Ervasti J.M. Dystrophin-glycoprotein complex is monomeric and stabilizes actin filaments in vitro through a lateral association. J. Biol. Chem. 1997;272:28771–28778.9353348Rybakova I.N., Amann K.J., Ervasti J.M. A new model for the interaction of dystrophin with F-actin. J. Cell Biol. 1996;135:661–672.PMC21210718909541Straub V., Campbell K.P. Muscular dystrophies and the dystrophin-glycoprotein complex. Curr. Opin. Neurobiol. 1997;10:168–175.9146999Straub V., Bittner R.E., Léger J.J., Voit T. Direct visualization of the dystrophin network on skeletal muscle fiber membrane. J. Cell Biol. 1992;119:1183–1191.PMC22897251447296Straub V., Rafael J.A., Chamberlain J.S., Campbell K.P. Animal models for muscular dystrophy show different patterns of sarcolemmal disruption. J. Cell Biol. 1997;139:375–385.PMC21397919334342Tinsley J., Deconinck N., Fisher R., Kahn D., Phelps S., Gillis J.M., Davies K. Expression of full-length utrophin prevents muscular dystrophy in mdx mice. Nature Med. 1998;4:1441–1444.9846586Williams M.W., Bloch R.J. Extensive but coordinated reorganization of the membrane skeleton in myofibers of dystrophic (mdx) mice. J. Cell Biol. 1999;144:1259–1270.PMC215059110087268