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Basolateral but not apical localization of sodium‐dependent vitamin C transporter 2 (SVCT2) in mouse small intestine epithelial cells
Author(s) -
Kuo ShiuMing,
Sigurdson Wade J
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1202.7
Subject(s) - small intestine , alexa fluor , biology , microbiology and biotechnology , polyclonal antibodies , immunofluorescence , antibody , chemistry , fluorescence , biochemistry , immunology , physics , quantum mechanics
EGFP‐tagged human SVCT2 was previously shown to be basolaterally localized in cultured intestinal epithelial cells, Caco‐2. To confirm the localization of endogenous SVCT2, we performed immunofluorescence staining of cryosections from mouse small intestine (duodenum and ileum) using polyclonal anti‐SVCT2 antibodies and Alexa Fluor® 488‐conjugated goat anti‐rabbit IgG as the secondary antibodies. To ensure the presence of proper reference points to the tissue/cell structure, carbocyanine dye, DiI, was used to concurrently label the whole section. Through two‐color confocal microscopic fluorescence imaging, we found that similar to Na,K‐ATPase (visualized by using rabbit anti‐Na+/K+ ATPase α IgG), SVCT2 was basolaterally localized and no apical localization was detected in the mouse intestinal epithelial cells. Consistent with the known wide‐spread SVCT2 expression in the body, SVCT2 was also found in other cell types in small intestine sections. SVCT2+/− mice showed greatly diminished SVCT2 expression in intestinal sections although the Na,K‐ATPase and DiI stainings of sections were comparable to that of the wildtype mice.