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
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Research Support, Non-U.S. Gov't
MeSH terms
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Actins / isolation & purification
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Base Sequence
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Carrier Proteins / genetics
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Carrier Proteins / physiology*
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Cell Compartmentation / physiology
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Cell Polarity / physiology*
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Clone Cells
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Cytoskeleton / physiology
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Down-Regulation
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Epithelial Cells
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Epithelium / drug effects
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Epithelium / physiology
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Epithelium / ultrastructure
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Humans
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Immunohistochemistry
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Intestines / cytology
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Intestines / drug effects
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Intestines / physiology*
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Intestines / ultrastructure
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Microfilament Proteins / genetics
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Microfilament Proteins / physiology*
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Microscopy, Electron
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Microvilli / drug effects
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Microvilli / physiology*
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Microvilli / ultrastructure
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Molecular Sequence Data
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Morphogenesis / drug effects
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RNA, Antisense / pharmacology*
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Sucrase-Isomaltase Complex / biosynthesis
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Sucrase-Isomaltase Complex / isolation & purification
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Suppression, Genetic
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Transfection
Substances
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Actins
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Carrier Proteins
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Microfilament Proteins
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RNA, Antisense
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villin
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Sucrase-Isomaltase Complex