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Human, rat and chicken small intestinal Na + ‐Cl − ‐creatine transporter: functional, molecular characterization and localization
Author(s) -
Peral M. J.,
GarcíaDelgado M.,
Calonge M. L.,
Durán J. M.,
Horra M. C.,
Wallimann T.,
Speer O.,
Ilundáin A. A.
Publication year - 2002
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1113/jphysiol.2002.026377
Subject(s) - creatine , biology , microbiology and biotechnology , small intestine , biochemistry , ileum , phosphocreatine , amino acid , endocrinology , energy metabolism
In spite of all the fascinating properties of oral creatine supplementation, the mechanism(s) mediating its intestinal absorption has(have) not been investigated. The purpose of this study was to characterize intestinal creatine transport. [ 14 C]Creatine uptake was measured in chicken enterocytes and rat ileum, and expression of the creatine transporter CRT was examined in human, rat and chicken small intestine by reverse transcription‐polymerase chain reaction, Northern blot, in situ hybridization, immunoblotting and immunohistochemistry. Results show that enterocytes accumulate creatine against its concentration gradient. This accumulation was electrogenic, Na + ‐ and Cl − ‐dependent, with a probable stoichiometry of 2 Na + : 1 Cl − : 1 creatine, and inhibited by ouabain and iodoacetic acid. The kinetic study revealed a K m for creatine of 29 μ m . [ 14 C]Creatine uptake was efficiently antagonized by non‐labelled creatine, guanidinopropionic acid and cyclocreatine. More distant structural analogues of creatine, such as GABA, choline, glycine, β‐alanine, taurine and betaine, had no effect on intestinal creatine uptake, indicating a high substrate specificity of the creatine transporter. Consistent with these functional data, messenger RNA for CRT was detected only in the cells lining the intestinal villus. The sequences of partial clones, and of the full‐length cDNA clone, isolated from human and rat small intestine were identical to previously cloned CRT cDNAs. Immunological analysis revealed that CRT protein was mainly associated with the apical membrane of the enterocytes. This study reports for the first time that mammalian and avian enterocytes express CRT along the villus, where it mediates high‐affinity, Na + ‐ and Cl − ‐dependent, apical creatine uptake.