Effects of HNK-1 monoclonal antibody on the substratum attachment and survival of neural crest and sclerotome cells in culture.
The sclerotome portion of the differentiating embryonic chick somite becomes infiltrated by neural crest cells prior to its dispersal towards the embryonic axis. This means that sclerotome cells explanted into culture for the purpose of examining their interactions in vitro are contaminated with a proportion of neural crest cells. The purpose of this study was to explore the neural crest cell adhesion epitope recognized by the HNK-1 monoclonal antibody and ways in which this antibody can be used to eliminate neural crest cells from mixed culture by selective cytotoxicity. Immunofluorescence technique, under the conditions used here, indicated that the antibody appeared to stain all the mesenchymal (i.e. neural crest) cells emigrating from pieces of embryonic neural tube in culture. Examination of the effects of HNK-1 suggests that the antibody interacts with substratum-binding sites on the neural crest cell surface. On fibronectin-coated substrata the antibody tended to inhibit neurite outgrowth but left the cells relatively well spread, while on laminin substrata the effect was to discourage both neurite extension and cell spreading, causing cell retraction. These results suggest that the cell surface epitope recognized by HNK-1 influences neurite outgrowth, neurite adhesiveness or both. Failure of cell spreading on laminin suggests interaction with the laminin binding sites on the cell body. Elimination of the crest cells from mixed culture with sclerotome was achieved by culturing the cells in the presence of HNK-1 antibody and complement during the period required for complete cell outgrowth from the sclerotome explant.(ABSTRACT TRUNCATED AT 250 WORDS)
Sanders
E J
EJ
Department of Physiology, University of Alberta, Edmonton, Canada.
Cheung
E
E
D000818:Animals; D000911:Antibodies, Monoclonal / Q000502:physiology; D002356:Cartilage / Q000196:embryology*; D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D005353:Fibronectins / Q000378:metabolism; D007797:Laminin / Q000378:metabolism; D008858:Microscopy, Phase-Contrast; D009432:Neural Crest / Q000166:cytology*
D000911:Antibodies, Monoclonal; D005353:Fibronectins; D007797:Laminin