Developmental modulation of embryonic cardiac myocyte adhesion to cardiac collagens in vitro.
The goal of this investigation is to identify molecules that mediate embryonic cardiac myocyte adhesion during chick cardiac morphogenesis. The assay used employs culturing embryonic myocytes on substrata containing embryonic heart proteins separated by molecular weight. This assay shows that embryonic myocytes from 10- to 14-day-old embryos will bind to 140,000 and 128,000 Da proteins present in embryonic hearts and do not require Mg2+ or Ca2+ for adhesion. Myocytes from embryos younger than 10 days or older than 14 days display little or no binding. Embryonic heart fibroblasts collected at these same ages do not bind to these proteins. The 140- and 128-kDa proteins were found to copurify in extraction procedures for procollagens. Amino acid analysis shows that both proteins contain high glycine and hydroxyproline, indicating that they are collagens. However, glycine and imino acid levels are low relative to other known collagens, indicating a nonhelical domain present in each molecule and most closely resembled levels present in procollagens. Immunoblots show that antisera to chick collagen type I recognizes the 128-kDa protein while anti-collagen type III recognizes the 140-kDa protein. Monoclonal antibodies to the amino terminal propeptide of collagen type I recognize the 128-kDa protein in immunoblotting procedures. Embryonic chick myocytes bind to 140/128 kDa proteins present in extracts of sympathetic trunk, although they do not bind to 140/128 kDa proteins in embryonic tendon. The findings thereby indicate that forms of type III and type I collagens in embryonic heart support direct adhesion of embryonic myocytes for a restricted period of cardiac myogenesis and that these proteins differ from collagen types I and III present in other tissues and from fully processed collagen types I and III.
Tidball
J G
JG
Department of Kinesiology, University of California, Los Angeles 90024.
Latus
L
L
Weckerle
J
J
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002448:Cell Adhesion; D002478:Cells, Cultured; D002642:Chick Embryo; D003094:Collagen / Q000378:metabolism*; D003893:Desmin / Q000276:immunology / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006321:Heart / Q000196:embryology*; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D003893:Desmin; D003094:Collagen