Carbohydrate deprivation reduces NADPH-production in fish liver but not in adipose tissue.
Little is known about the way in which carnivorous fish such as salmonids mobilise and metabolise dietary carbohydrates, which are essential to lipid metabolism. Thus we have studied changes caused by the absence of dietary carbohydrates to the kinetics and molecular behaviour of the four cellular NADPH-production systems [glucose 6-phosphate dehydrogenase (G6PDH); 6-phosphogluconate dehydrogenase (6PGDH); malic enzyme (ME); and isocitrate dehydrogenase NADP-dependent (NADP-IDH)] in the liver and adipose tissue of rainbow trout (Oncorhynchus mykiss). We used spectrophotometry to study enzyme kinetics and nucleic acid concentrations, and immunoblot analysis to determine specific protein concentrations. The absence of carbohydrate reduced specific enzyme activity, maximum rate and catalytic efficiency by almost 65% in G6PDH and 6PGDH, by more than 50% in ME, and by almost 25% in NADP-IDH but caused no significant changes in the K(m) values or activity ratios in any of these hepatic enzymes. Molecular analysis clearly showed that this kinetic behaviour reflected concomitant changes in intracellular enzyme concentrations, produced by protein-induction/repression processes rather than changes in the activity of pre-existing enzymes. We conclude that the absence of carbohydrates significantly reduces intracellular concentrations of G6PDH, ME and NADP-IDH in trout liver in percentages similar to those recorded for enzyme activity. We found no such variations in the concentrations of any of these enzymes in adipose tissue and no change in the levels of their activity, suggesting that the liver and adipose tissues are subject to different regulation systems with regard to carbohydrates and play distinct roles in lipid metabolism.
Barroso
J B
JB
Department of Biochemistry and Molecular Biology, Faculty of Experimental Sciences, University of Jaén, 23071 Jaén, Spain.
Peragón
J
J
García-Salguero
L
L
de la Higuera
M
M
Lupiáñez
J A
JA
D000273:Adipose Tissue / Q000201:enzymology*; D000818:Animals; D004040:Dietary Carbohydrates / Q000008:administration & dosage* / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D015151:Immunoblotting; D007521:Isocitrate Dehydrogenase / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008291:Malate Dehydrogenase / Q000378:metabolism; D009249:NADP / Q000096:biosynthesis*; D017686:Oncorhynchus mykiss / Q000378:metabolism*; D010734:Phosphogluconate Dehydrogenase / Q000378:metabolism
D004040:Dietary Carbohydrates; D009249:NADP; D008291:Malate Dehydrogenase; D007521:Isocitrate Dehydrogenase; C039355:isocitrate dehydrogenase (NADP+); D010734:Phosphogluconate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase