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1 College of Pharmacy and
Upjohn Center for Clinical Pharmacology,
2 Department of Physiology and
3 Division of Nephrology,
To determine the
renal localization of oligopeptide transporters, Northern
blot analyses were performed and polyclonal antisera were generated
against PEPT1 and PEPT2, the two cloned rat
H+/peptide transporters. Under
high-stringency conditions, a 3.0-kb mRNA transcript of rat PEPT1 was
expressed primarily in superficial cortex, whereas a 3.5-kb mRNA
transcript of PEPT2 was expressed primarily in deep cortex/outer stripe
of outer medulla. PEPT1 antisera detected a specific band on
immunoblots of renal and intestinal brush-border membrane vesicles
(BBMV) with an apparent mobility of ~90 kDa. PEPT2 antisera detected
a specific broad band of ~85 kDa in renal but not in intestinal BBMV.
PEPT1 immunolocalization experiments showed detection of a brush border
antigen in S1 segments of the proximal tubule and in the brush border
of villi from all segments of the small intestine. In contrast, PEPT2
immunolocalization was primarily confined to the brush border of S3
segments of the proximal tubule. All other nephron segments in rat were
negative for PEPT1 and PEPT2 staining. Overall, our results
conclusively demonstrate that although PEPT1 is expressed in early
regions of the proximal tubule (pars convoluta), PEPT2 is specific for the latter regions of proximal tubule (pars recta).
Northern blot analysis; immunoblots; immunocytochemistry; proximal tubules
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