Analysis of tissue-specific and PPARalpha-dependent induction of FABP gene expression in the mouse liver by an in vivo DNA electroporation method.
Peroxisome proliferator (PPAR)alpha ligand Wy14,643 induces liver-fatty acid binding protein (FABP) spontaneously and heart-FABP gradually, but not intestine-FABP mRNA expression in the mouse liver. These strict regulations have not been reproduced in cultured cell systems. We applied a DNA electroporation method to directly introduce reporter gene constructs into the livers of mice. This system reproduced the in vivo responses of the above three FABP gene promoters to the PPARalpha ligand but not that of a promoter containing the typical three PPAR binding sites in tandem. Deletion and mutation analyses of the mouse L-FABP gene suggested that, in addition to the binding site for PPARalpha, a far upstream sequence is required for PPAR-dependent transactivation in the liver. In contrast to the cultured cell systems, our in vivo DNA electroporation method showed that PPARalpha binding to the promoter is necessary but not sufficient for PPARalpha ligand-dependent transcriptional activation of the L-FABP gene in vivo.
Fujishiro
Kensei
K
Department of Biochemistry, School of Pharmaceutical Sciences, Toho University, Funabashi, Chiba, Japan.
Fukui
Yuka
Y
Sato
Osamu
O
Kawabe
Kohei
K
Seto
Koichi
K
Motojima
Kiyoto
K
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D018274:Electroporation / Q000379:methods*; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D008099:Liver / Q000502:physiology*; D008297:Male; D051379:Mice; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D014018:Tissue Distribution; D014157:Transcription Factors / Q000378:metabolism*
D002352:Carrier Proteins; C493320:Fabp1 protein, mouse; C082667:Fabp5 protein, mouse; C086936:Fabp7 protein, mouse; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors; D004247:DNA