Organization of intestinal epithelial cells into multicellular structures requires laminin and functional actin microfilaments.
Epithelial cell organization into multicellular structures is a critical biological process required for both organogenesis and repair following injury. The basement membrane and the cytoskeleton have important roles in this process; however, the functions of individual components of basement membrane and cytoskeleton are poorly understood. We used IEC-6 cells, a rat intestinal crypt cell line, grown on a three-dimensional gel of reconstituted basement membrane as a model system to determine which extracellular matrix and cytoskeletal components mediate intestinal epithelial cell organization. The cells entered the gel and formed hollow, tubular structures that resembled intestinal crypts. These structures were characterized by a single layer of polarized cells with apical tight junctions and microvilli on the luminal surface. Antiserum to laminin and the pentapeptide Tyr-Ile-Gly-Ser-Arg (which prevents cell attachment to laminin) inhibited this organization, but a control pentapeptide (Tyr-Tyr-Gly-Asp-Ala) and antiserum to collagen IV did not. Cytochalasin B, which interferes with actin microfilament polymerization, also inhibited organization of cells into multicellular structures, but vinblastine and Colcemid, which disrupt microtubules, and cycloheximide, which inhibits protein synthesis, did not. We conclude that organization of intestinal epithelial cells on a basement membrane into multicellular structures results from specific interactions between cells and laminin and requires intact actin microfilaments.
Olson
A D
AD
Department of Pediatrics, Schneider Children's Hospital, Long Island Jewish Medical Center, New Hyde Park, New York 11042.
Pysher
T
T
Bienkowski
R S
RS
D008841:Actin Cytoskeleton / Q000502:physiology*; D000199:Actins / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D001485:Basement Membrane / Q000502:physiology; D002448:Cell Adhesion; D003094:Collagen / Q000502:physiology; D003599:Cytoskeleton / Q000502:physiology; D007413:Intestinal Mucosa / Q000166:cytology*; D007797:Laminin / Q000502:physiology*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000378:metabolism; D051381:Rats; D016212:Transforming Growth Factor beta / Q000502:physiology
D000199:Actins; D007797:Laminin; D010446:Peptide Fragments; D016212:Transforming Growth Factor beta; D003094:Collagen