A hyaluronan-binding protein shows a partial and temporally regulated codistribution with actin on locomoting chick heart fibroblasts.
The ultrastructural distribution of a hyaluronan-binding protein (HABP) and its relationship to actin-containing microfilaments were studied with immunocytochemistry. Ultrastructural analysis localized HABP to the cell coat and demonstrated that it occurred largely in cell processes where the apical surfaces were immunopositive. The codistribution of HABP with actin-containing microfilaments in cell processes was demonstrated with double immunolabeling using monoclonal antibodies to actin and monospecific, polyclonal antibodies to HABP. Both the topological localization of HABP and its cytoskeletal coassociations were modulated by cells during different cellular phases. Thus, in cells exhibiting large lamellae and few actin fibrils, typical of rapidly locomoting cells, HABP codistributed primarily with the actin meshwork occurring in cell processes, although some codistribution between the two proteins occurred over the cell body. In cells containing prominent stress fibers and less obvious lamellae, HABP was absent in cell processes but, rather, was aligned primarily along actin fibrils occurring in the cell body. A functional association between HABP and the actin-containing cytoskeleton was suggested by the ability of cytochalasin D to coordinately alter the distribution of HABP and disrupt its coassociation with actin. As well, the addition of hyaluronan to monolayers increased the association of HABP with a Triton-insoluble cytoskeleton. The possible roles of HABP in cell motility and cytoskeletal organization are discussed.
Department of Pharmacology, University of Calgary Health Sciences Centre, Alberta, Canada.
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Turley
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Brassel
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Moore
D008841:Actin Cytoskeleton / Q000648:ultrastructure*; D000199:Actins / Q000032:analysis*; D000818:Animals; D002352:Carrier Proteins / Q000032:analysis*; D002465:Cell Movement; D002478:Cells, Cultured; D002642:Chick Embryo; D003599:Cytoskeleton / Q000648:ultrastructure*; D005318:Fetal Heart / Q000502:physiology*; D005347:Fibroblasts / Q000502:physiology / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid / Q000378:metabolism; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D009206:Myocardium / Q000166:cytology / Q000648:ultrastructure; D009924:Organ Culture Techniques
D000199:Actins; D002352:Carrier Proteins; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid