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

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 mess...

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Bibliographic Details
Published in:	The EMBO journal Vol. 14; no. 3; pp. 409 - 421
Main Authors:	Costa de Beauregard, M.A., Pringault, E., Robine, S., Louvard, D.
Format:	Journal Article
Language:	English
Published:	England 01-02-1995
Subjects:	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
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Summary:	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.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23
ISSN:	0261-4189 1460-2075
DOI:	10.1002/j.1460-2075.1995.tb07017.x