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Content of ileal EAAC1 and hepatic GLT-1 high-affinity glutamate transporters is increased in growing vs. nongrowing lambs, paralleling increased tissue D- and L-glutamate, plasma glutamine, and alanine concentrations¹

J. A. Howell^*, A. D. Matthews^*, T. C. Welbourne and J. C. Matthews^*,2

^* Department of Animal Sciences, University of Kentucky, Lexington 40546 and and Department of Molecular and Cellular Physiology, Louisiana State University Medical Center, Shreveport 71130

² Correspondence:
808 W. P. Garrigus Bldg. (phone: 859-257-7513; fax: 859-257-3412; E-mail:
jmatthew{at}uky.edu).

Glutamate is a central metabolite for whole-animal energy and N metabolism. This study tested the hypothesis that ileal epithelium, liver, and kidney content of system X^-_AG glutamate transporters EAAC1 and GLT-1 would be up-regulated to support growth of wethers (30 ± 1.2 kg) fed a forage-based diet for at least 14 d to gain (2.0 x NE_m; n = 9) vs. maintain (1.2 x NE_m; n = 9) BW. We have previously demonstrated that two high-affinity glutamate transporters (EAAC1, GLT-1) are expressed by these extensive glutamate metabolizing epithelial tissues. Wethers fed at 2.0 x NE_m gained (P < 0.001; 0.26 kg/d) BW, whereas those fed 1.2 x NE_m did not. Although plasma concentrations (µM) of glucose and L- or D-glutamate did not differ, plasma glutamine (precursor of glutamate) and alanine concentrations (transamination product of glutamate) were 28% (P < 0.007) and 22% (P < 0.072) greater for growing lambs than nongrowing lambs. In tissues, the concentration of L-glutamate in ileum epithelia and D-glutamate of liver was 49% (P < 0.015) and 181% (P < 0.042) greater, respectively, in growing vs. nongrowing animals, whereas concentrations of glutamate isoforms did not differ in kidney. Paralleling these increased amino acid concentrations, ileal epithelium contained 313% more (P < 0.038) EAAC1 protein and liver contained 240% more (P < 0.001) GLT-1 protein, whereas kidney transporter content did not differ between growing and nongrowing wethers. In contrast to increased EAAC1 and GLT-1 protein content in ileal and liver tissue of growing lambs, messenger RNA levels did not differ. These results indicate that the increased capacity for high-affinity glutamate uptake in growing vs. nongrowing lambs is achieved through increased expression of EAAC1 by ileal epithelium and GLT1 by liver, which parallel increased tissue concentrations of glutamate and plasma concentrations of two major interorgan N carriers, glutamine and alanine.

Key Words: Amino Acids • Biochemical Transport • Glutamates • Growth • Sheep

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