Differential expression and targeting of endogenous Arf1 and Arf6 small GTPases in kidney epithelial cells in situ

Jaafar El-Annan, Dennis Brown, Sylvie Breton, Sylvain Bourgoin, Dennis A. Ausiello, Vladimir Marshansky

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

ADP-ribosylation factors (Arfs) are small GTP-ases that regulate vesicular trafficking in exo- and endocytotic pathways. As a first step in understanding the role of Arfs in renal physiology, immunocytochemistry and Western blotting were performed to characterize the expression and targeting of Arf1 and Arf6 in epithelial cells in situ. Arf1 and Arf6 were associated with apical membranes and subapical vesicles in proximal tubules, where they colocalized with megalin. Arf1 was also apically expressed in the distal tubule, connecting segment, and collecting duct (CD). Arf1 was abundant in intercalated cells (IC) and colocalized with V-ATPase in A-IC (apical) and B-IC (apical and/or basolateral). In contrast, Arf6 was associated exclusively with basolateral membranes and vesicles in the CD. In the medulla, basolateral Arf6 was detectable mainly in A-IC. Expression in principal cells became weaker throughout the outer medulla, and Arf6 was not detectable in principal cells in the inner medulla. In some kidney epithelial cells Arf1 but not Arf6 was also targeted to a perinuclear patch, where it colocalized with TGN38, a marker of the trans-Golgi network. Quantitative Western blotting showed that expression of endogenous Arf1 was 26-180 times higher than Arf6. These data indicate that Arf GTPases are expressed and targeted in a cell- and membrane-specific pattern in kidney epithelial cells in situ. The results provide a framework on which to base and interpret future studies on the role of Arf GTPases in the multitude of cellular trafficking events that occur in renal tubular epithelial cells.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume286
Issue number4 55-4
StatePublished - Apr 2004
Externally publishedYes

Fingerprint

Monomeric GTP-Binding Proteins
Epithelial Cells
ADP-Ribosylation Factors
Kidney
GTP Phosphohydrolases
Membranes
Ducts
Low Density Lipoprotein Receptor-Related Protein-2
Physiology
Guanosine Triphosphate
Western Blotting
trans-Golgi Network
Adenosine Triphosphatases
Immunohistochemistry
Cell Membrane

Keywords

  • Endocytosis
  • Immunofluorescence microscopy
  • Protein trafficking
  • Western blotting

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Differential expression and targeting of endogenous Arf1 and Arf6 small GTPases in kidney epithelial cells in situ. / El-Annan, Jaafar; Brown, Dennis; Breton, Sylvie; Bourgoin, Sylvain; Ausiello, Dennis A.; Marshansky, Vladimir.

In: American Journal of Physiology - Cell Physiology, Vol. 286, No. 4 55-4, 04.2004.

Research output: Contribution to journalArticle

El-Annan, Jaafar ; Brown, Dennis ; Breton, Sylvie ; Bourgoin, Sylvain ; Ausiello, Dennis A. ; Marshansky, Vladimir. / Differential expression and targeting of endogenous Arf1 and Arf6 small GTPases in kidney epithelial cells in situ. In: American Journal of Physiology - Cell Physiology. 2004 ; Vol. 286, No. 4 55-4.
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