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Am J Physiol Regul Integr Comp Physiol 254: R801-R808, 1988;
0363-6119/88 $5.00
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AJP - Regulatory, Integrative and Comparative Physiology, Vol 254, Issue 5 801-R808, Copyright © 1988 by American Physiological Society


ARTICLES

Characteristics of L-alanine uptake in freshly isolated hepatocytes of elasmobranch Raja erinacea

N. Ballatori and J. L. Boyer
Mount Desert Island Biological Laboratory, Salsbury Cove, Maine 04672.

The specific transport mechanisms that mediate the hepatic uptake of L-[3H]alanine and of an unnatural homologue, alpha-[14C]methylaminoisobutyric acid (MeAIB), were analyzed in hepatocyte suspensions from Raja erinacea. Aminooxyacetate, an inhibitor of aminotransferase activity was used to prevent alanine metabolism. After 3 h of incubation with either 0.5 mM alanine or MeAIB, hepatic concentrations of these amino acids were significantly higher in the presence than absence of Na+ (8 vs. 1 and 1 vs. 0.1 mM, respectively). Kinetic studies indicated that both alanine and MeAIB transport occurred via sodium-dependent saturable mechanisms. [14C]MeAIB uptake was completely inhibited by excess L-alanine. Uptake of [3H]alanine was inhibited by a 40-fold excess of serine and cysteine (53-54%), by MeAIB and methylalanine (26-31%), and by leucine (14%), whereas D-alanine, beta-alanine, taurine, and glutamate had no effect. Insulin and glucagon were unable to stimulate [3H]alanine uptake. Glucose release from hepatocytes was unaffected by 10 mM alanine or 2 mM aminooxyacetate, indicating that alanine is not a major gluconeogenic precursor in this marine elasmobranch. These results suggest that uptake of L-alanine by skate hepatocytes occurs predominantly via a sodium-dependent system, with properties similar to those exhibited by the ASC neutral amino acid transport system previously characterized in Ehrlich ascites tumor cells and rat hepatocytes.


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