A study of the contribution of minor GABA metabolites to the control of feeding in the rat
This project aimed to investigate the role of the γ-lactone of 3,4-dihydroxybutanoic acid (3,4DB) which was claimed by Japanese investigators to be one of several endogenous γ-lactones involved in the control of food intake. Tissue, plasma and urine organic acid profiles were screened for the γ-lactone of 3,4DB using both GC and GCMS. Careful mass spectral analysis and in vitro acid-γ-lactone exchange analyses with structural validation studies showed that the γ-lactone of 3,4DB did not occur in vivo but that the free acid of 3,4DB did. This rejects previous claims to the contrary. Fasting increased rat plasma 3,4DB with urinary output significantly elevated for the first 24h of fasting. A metabolic route from glutamate to acetate is proposed with 3,4DB as an intermediate. Peripheral administration of glutamate, γ-aminobutyric acid (GABA) and 4-hydroxybutyric acid (4HB) to rats reduced food intake with increasing effects at each stage of the pathway. The γ-lactone 2-buten-4-olide is toxic and other γ-lactones had more potent effects than their free acids on rats. The intragastric administration of GABA stimulated 4HB production in vivo and GABA or 4HB increased plasma 3,4DB levels, thus implying that 3,4DB occurs at an intermediate in our proposed pathway. A likely endogenous source of 3,4DB is as a minor GABA metabolite. The response of the free acid of 3,4DB to fasting followed a similar trend as that reported for its γ-lactone and related γ-lactones are unlikely to occur in vivo. Given the similarities of 4HB (a precursor of 3,4DB) and 3,4DB in structure to the ketone body 3-hydroxybutyrate (3HB) and the similarity of 3,4DB in fasting response to 3HB, both 4HB and 3,4DB may initiate anorexia in fasting.