In vitro studies on adult cardiac myocytes: attachment and biosynthesis of collagen type IV and laminin.
The interactions between adult rat cardiac myocytes and the basement membrane components collagen type IV and laminin were investigated in attachment experiments and biosynthesis studies and by immunofluorescence staining. Adult myocytes attached equally well to native collagen type IV and laminin but did not attach to collagen type IV solubilized with pepsin (P-CIV) or to collagen type I. However, when laminin was used to coat P-CIV, attachment was enhanced. Affinity-purified antibodies against laminin inhibited the attachment of myocytes to dishes coated with native collagen type IV, indicating that cell surface-bound laminin mediated attachment of the cells to this substrate. Immunofluorescence staining of freshly isolated myocytes, using antibodies against laminin or collagen type IV, revealed the presence of laminin but not of collagen type IV on the surface of freshly isolated cells, indicating that during the isolation procedure collagen IV was removed from the cell surface. Metabolic labeling followed by immunoprecipitation demonstrated synthesis of both laminin and collagen type IV in cardiac myocytes as they progressed into culture over a 14-day period. This synthesis was accompanied by the deposition of the collagen type IV and laminin into distinctly different patterns as revealed by immunofluorescence staining. As the cells progressed into culture, newly synthesized laminin formed a network radiating from the center of the reorganizing cell into the pseudopods. The laminin was redistributed and remodeled with time in culture to form a dense layer beneath the cell. Collagen type IV was also synthesized with time in culture, but the pattern was a much finer network as opposed to the denser pattern of laminin staining. These studies demonstrate that adult cardiac myocytes synthesize and remodel the basement membrane as they adapt to the culture environment.
Lundgren
E
E
Department of Medical and Physiological Chemistry, Uppsala University, Sweden.
Gullberg
D
D
Rubin
K
K
Borg
T K
TK
Terracio
M J
MJ
Terracio
L
L
D000328:Adult; D002448:Cell Adhesion; D003094:Collagen / Q000096:biosynthesis*; D005455:Fluorescent Antibody Technique; D006801:Humans; D066298:In Vitro Techniques; D007797:Laminin / Q000096:biosynthesis*; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism
D007797:Laminin; D003094:Collagen