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Journal of Histochemistry and Cytochemistry, Vol. 48, 1441-1452, November 2000, Copyright © 2000, The Histochemical Society, Inc.

ARTICLE

Heterogeneous Distribution of Isoactins in Cultured Vascular Smooth Muscle Cells Does Not Reflect Segregation of Contractile and Cytoskeletal Domains

Jian Songa, Nathalie F. Wortha, Barbara E. Rolfea, Gordon R. Campbella, and Julie H. Campbella
a Centre for Research in Vascular Biology, Department of Anatomical Sciences, University of Queensland, Brisbane, Australia

Correspondence to: Julie H. Campbell, Centre for Research in Vascular Biology, Dept. of Anatomical Sciences, The University of Queensland, Brisbane QLD 4072, Australia. E-mail: julie.campbell@mailbox.uq.edu.au

We have previously demonstrated that {alpha}-smooth muscle ({alpha}-SM) actin is predominantly distributed in the central region and ß-non-muscle (ß-NM) actin in the periphery of cultured rabbit aortic smooth muscle cells (SMCs). To determine whether this reflects a special form of segregation of contractile and cytoskeletal components in SMCs, this study systematically investigated the distribution relationship of structural proteins using high-resolution confocal laser scanning fluorescent microscopy. Not only isoactins but also smooth muscle myosin heavy chain, {alpha}-actinin, vinculin, and vimentin were heterogeneously distributed in the cultured SMCs. The predominant distribution of ß-NM actin in the cell periphery was associated with densely distributed vinculin plaques and disrupted or striated myosin and {alpha}-actinin aggregates, which may reflect a process of stress fiber assembly during cell spreading and focal adhesion formation. The high-level labeling of {alpha}-SM actin in the central portion of stress fibers was related to continuous myosin and punctate {alpha}-actinin distribution, which may represent the maturation of the fibrillar structures. The findings also suggest that the stress fibers, in which actin and myosin filaments organize into sarcomere-like units with {alpha}-actinin-rich dense bodies analogous to Z-lines, are the contractile structures of cultured SMCs that link to the network of vimentin-containing intermediate filaments through the dense bodies and dense plaques. (J Histochem Cytochem 48:1441–1452, 2000)

Key Words: actin isoform, myosin heavy chain, vimentin, {alpha}-actinin, vinculin


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