Suppression of villin expression by antisense RNA impairs brush border assembly in polarized epithelial intestinal cells

EMBO J. 1995 Feb 1;14(3):409-21.

Abstract

We have used an antisense RNA strategy to investigate the role of the actin-associated protein, villin, in the brush-border morphogenesis of human intestinal CaCO2 cells. Stable expression of a cDNA encoding antisense villin RNA resulted in the permanent down-regulation of the endogenous villin message and dramatically affected brush-border assembly. Ultrastructural and immunolocalization studies revealed that epithelial cell polarity was largely maintained. However, in contrast to brush-border markers such as dipeptidyl-peptidase IV, the apical localization of sucrase-isomaltase was specifically impaired. Retransfection of the villin antisense-expressing cell line with a cDNA encoding a partial sense villin RNA restored both brush-border assembly and sucrase-isomaltase apical expression. The suggestion that brush-border morphogenesis may be important for the trafficking of certain proteins is discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / isolation & purification
  • Base Sequence
  • Carrier Proteins / genetics
  • Carrier Proteins / physiology*
  • Cell Compartmentation / physiology
  • Cell Polarity / physiology*
  • Clone Cells
  • Cytoskeleton / physiology
  • Down-Regulation
  • Epithelial Cells
  • Epithelium / drug effects
  • Epithelium / physiology
  • Epithelium / ultrastructure
  • Humans
  • Immunohistochemistry
  • Intestines / cytology
  • Intestines / drug effects
  • Intestines / physiology*
  • Intestines / ultrastructure
  • Microfilament Proteins / genetics
  • Microfilament Proteins / physiology*
  • Microscopy, Electron
  • Microvilli / drug effects
  • Microvilli / physiology*
  • Microvilli / ultrastructure
  • Molecular Sequence Data
  • Morphogenesis / drug effects
  • RNA, Antisense / pharmacology*
  • Sucrase-Isomaltase Complex / biosynthesis
  • Sucrase-Isomaltase Complex / isolation & purification
  • Suppression, Genetic
  • Transfection

Substances

  • Actins
  • Carrier Proteins
  • Microfilament Proteins
  • RNA, Antisense
  • villin
  • Sucrase-Isomaltase Complex