Ontogeny and subcellular localization of rat liver mitochondrial branched chain amino-acid aminotransferase.
Branched chain amino-acid aminotransferase (BCAT) activity is present in fetal liver but the developmental pattern of mitochondrial BCAT (BCATm) expression in rat liver has not been studied. The aim of this study was to determine the activity, protein and mRNA concentration of BCATm in fetal and postnatal rat liver, and to localize this enzyme at the cellular and subcellular levels at both developmental stages. Maximal BCAT activity and BCATm mRNA expression occurred at 17 days' gestation in fetal rat liver and then declined significantly immediately after birth. This pattern was observed only in liver; rat heart showed a different developmental pattern. Fetal liver showed intense immunostaining to BCATm in the nuclei and mitochondria of hepatic cells and blood cell precursors; in contrast, adult liver showed mild immunoreactivity located only in the mitochondria of hepatocytes. BCAT activity in isolated fetal liver nuclei was 0.64 mU x mg(-1) protein whereas it was undetectable in adult liver nuclei. By Western blot analysis the BCATm antibody recognized a 41-kDa protein in fetal liver nuclei, and proteins of 41 and 43 kDa in fetal liver supernatant. In adult rat liver supernatant, the BCATm antibody recognized only a 43-kDa protein; however, neither protein was detected in adult rat liver nuclei. The appearance of the 41-kDa protein was associated with the presence of the highly active form of BCATm. These results suggest the existence of active and inactive forms of BCAT in rat liver.
Torres
N
N
Departamento de Fisiología de la Nutrición, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, México.
Vargas
C
C
Hernández-Pando
R
R
Orozco
H
H
Hutson
S M
SM
Tovar
A R
AR
D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000201:enzymology; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D005318:Fetal Heart / Q000201:enzymology; D005333:Fetus / Q000201:enzymology; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D006321:Heart / Q000254:growth & development; D007150:Immunohistochemistry; D008099:Liver / Q000196:embryology / Q000201:enzymology / Q000254:growth & development; D008297:Male; D008930:Mitochondria, Liver / Q000201:enzymology*; D009206:Myocardium / Q000201:enzymology; D010920:Placenta / Q000201:enzymology; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013347:Subcellular Fractions / Q000201:enzymology; D000637:Transaminases / Q000235:genetics / Q000378:metabolism*
D018076:DNA, Complementary; D012333:RNA, Messenger; D000637:Transaminases; C019798:branched-chain-amino-acid transaminase