Evolution of pyruvate carboxylase and other biotin containing enzymes in developing rat liver and kidney.
The evolution of pyruvate carboxylase has been studied in rat liver and kidney during perinatal development. The pyruvate carboxylase activity, amount of enzyme and mRNA levels have been assayed from 2 days before delivery to weaning. In liver, there is a peak of activity and amount of enzyme 24 h before delivery and 2 peaks, at 12 h and 6 days, after parturition. The transcription of the enzyme gene followed a similar pattern, with mRNA peaks preceding those of activity and amount of enzyme. However, in kidney, pyruvate carboxylase activity, amount and mRNA remain low until weaning. These results confirm the limited role of renal gluconeogenesis during the perinatal development. Since all carboxylases contain biotin as prosthetic group, the biotinylation of pyruvate carboxylase during the perinatal period was investigated by western-blot using streptavidin-biotin peroxidase. In the mitochondrial samples from liver and kidney, all the pyruvate carboxylase detected was fully biotinylated, indicating an early development of the holocarboxylase synthetase activity in the perinatal period. This Western-blot technique also allowed us the detection of other biotin-enzymes based on their molecular weight. In liver, during the perinatal development propionyl-coA and 3-methyl-crotonyl-coA carboxylases followed a pattern of induction similar to pyruvate carboxylase. In kidney, the expression of mitochondrial carboxylases was lower compared to liver and propionyl-coA carboxylase was not detected during the studied period.
Salto
R
R
Departamento de Bioquímica y Biología Molecular, Facultad de Farmacia, Universidad de Granada, Campus de Cartuja, Spain.
Girón
M D
MD
del Mar Sola
M
M
Vargas
A M
AM
D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D001710:Biotin / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005333:Fetus / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D007668:Kidney / Q000196:embryology / Q000201:enzymology* / Q000254:growth & development*; D008099:Liver / Q000196:embryology / Q000201:enzymology* / Q000254:growth & development*; D008297:Male; D008928:Mitochondria / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology; D011247:Pregnancy; D011766:Pyruvate Carboxylase / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
D017931:DNA Primers; D012333:RNA, Messenger; D001710:Biotin; D011766:Pyruvate Carboxylase