CFTR channel insertion to the apical surface in rat duodenal villus epithelial cells is upregulated by VIP in vivo

Nadia A. Ameen, Birgitta Martensson, Lilly Bourguinon, Christopher Marino, Jon Isenberg, Gwenn E McLaughlin

Research output: Contribution to journalArticle

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Abstract

cAMP activated insertion of the cystic fibrosis transmembrane conductance regulator (CFTR) channels from endosomes to the apical plasma membrane has been hypothesized to regulate surface expression and CFTR function although the physiologic relevance of this remains unclear. We previously identified a subpopulation of small intestinal villus epithelial cells or CFTR high expressor (CHE) cells possessing very high levels of apical membrane CFTR in association with a prominent subapical vesicular pool of CFTR. We have examined the subcellular redistribution of CFTR in duodenal CHE cells in vivo in response to the cAMP activated secretagogue vasoactive intestinal peptide (VIP). Using anti-CFTR antibodies against the C terminus of rodent CFTR and indirect immunofluorescence, we show by quantitative confocal microscopy that CFTR rapidly redistributes from the cytoplasm to the apical surface upon cAMP stimulation by VIP and returns to the cytoplasm upon removal of VIP stimulation of intracellular cAMP levels. Using ultrastructural and confocal immunofluorescence examination in the presence or absence of cycloheximide, we also show that redistribution was not dependent on new protein synthesis, changes in endocytosis, or rearrangement of the apical cytoskeleton. These observations suggest that physiologic cAMP activated apical membrane insertion and recycling of CFTR channels in normal CFTR expressing epithelia contributes to the in vivo regulation of CFTR mediated anion transport.

Original languageEnglish
Pages (from-to)887-894
Number of pages8
JournalJournal of Cell Science
Volume112
Issue number6
StatePublished - Apr 26 1999

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Cystic Fibrosis Transmembrane Conductance Regulator
Vasoactive Intestinal Peptide
Epithelial Cells
Cytoplasm
Membranes
Endosomes
Recycling
Cycloheximide
Indirect Fluorescent Antibody Technique
Endocytosis
Cytoskeleton
Confocal Microscopy
Fluorescent Antibody Technique
Anions
Rodentia
Epithelium

Keywords

  • CFTR
  • Channel insertion
  • Small intestine
  • VIP

ASJC Scopus subject areas

  • Cell Biology

Cite this

CFTR channel insertion to the apical surface in rat duodenal villus epithelial cells is upregulated by VIP in vivo. / Ameen, Nadia A.; Martensson, Birgitta; Bourguinon, Lilly; Marino, Christopher; Isenberg, Jon; McLaughlin, Gwenn E.

In: Journal of Cell Science, Vol. 112, No. 6, 26.04.1999, p. 887-894.

Research output: Contribution to journalArticle

Ameen, NA, Martensson, B, Bourguinon, L, Marino, C, Isenberg, J & McLaughlin, GE 1999, 'CFTR channel insertion to the apical surface in rat duodenal villus epithelial cells is upregulated by VIP in vivo', Journal of Cell Science, vol. 112, no. 6, pp. 887-894.
Ameen, Nadia A. ; Martensson, Birgitta ; Bourguinon, Lilly ; Marino, Christopher ; Isenberg, Jon ; McLaughlin, Gwenn E. / CFTR channel insertion to the apical surface in rat duodenal villus epithelial cells is upregulated by VIP in vivo. In: Journal of Cell Science. 1999 ; Vol. 112, No. 6. pp. 887-894.
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