EAAT 1 is involved in transport of L-glutamate during differentiation of the Caco-2 cell line

Mordrelle, Agnès, Eric Jullian, Cyrille Costa, Estelle Cormet-Boyaka, Robert Benamouzig, Daniel Tomé, and Jean-François Huneau. EAAT1 is involved in transport of L-glutamate during differentiation of the Caco-2 cell line. Am J Physiol Gastrointest Liver Physiol 279: G366–G373, 2000.—Little is known concerning the expression of amino acid transporters during intestinal epithelial cell differentiation. The transport mechanism of L-glutamate and its regulation during the differentiation process were investigated using the human intestinal Caco-2 cell line. Kinetic studies demonstrated the presence of a single, highaffinity, D-aspartate-sensitive L-glutamate transport system in both confluent and fully differentiated Caco-2 cells. This transport was clearly Na dependent, with a Hill coefficient of 2.9 6 0.3, suggesting a 3 Na-to-1 glutamate stoichiometry and corresponding to the well-characterized XA,G 2 system. The excitatory amino acid transporter (EAAT)1 transcript was consistently expressed in the Caco-2 cell line, whereas the epithelial and neuronal EAAT3 transporter was barely detected. In contrast with systems B and y, which have previously been reported to be downregulated when Caco-2 cells stop proliferating, L-glutamate transport capacity was found to increase steadily between day 8 and day 17. This increase was correlated with the level of EAAT1 mRNA, which might reflect an increase in EAAT1 gene transcription and/or stabilization of the EAAT1 transcript.

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