Differential expression of glutathione reductase and cytosolic glutathione peroxidase, GPX1, in developing rat lungs and kidneys.
A mutant rat GPX1 (a cytosolic predominant form), in which the selenocysteine residue in the catalytic center was replaced by cysteine, was prepared and an antibody against the mutant enzyme was raised. The resultant antibody specifically reacted with rat GPX1 and was, together with the Glutathione reductase (GR) antibody, used in a Western blot analysis and immunohistochemistry experiments. To elucidate the physiological coupling of these enzymes under oxidative stress which accompanies the birth, developmental changes of the protein levels and enzymatic activities of GR and GPX1 were examined for lungs and kidneys from prenatal fetus to adult rats. The expression of GR was already evident at the prenatal stage and remained high in lungs at all stages. However, GR activity in kidneys gradually increased after birth reaching maximal levels at adulthood. An immunohistochemical study showed that GR was strongly bound to the bronchial epithelia in lungs and the epithelial cells of renal tubes. GPX1 was expressed in the renal tube epithelial cells and its level gradually increased after birth in a manner similar to that of GR. The expression of GPX1 in the lungs was, on the other hand, variable and occurred in some alveolar cells and bronchial epithelia only at restricted periods. It preferentially localized in nuclei at a late stage of development. Thus, the expression of the two functionally coupled enzymes via GSH did not appear to coordinate with development, tissue localization or under oxidative stress. Since many gene products show GSH-dependent preoxidase activity, other peroxidase(s) may be induced to compensate for the low GPX1 levels at stages with high GR expression.
Fujii
Tsuneko
T
Department of Biochemistry, Osaka University Medical School, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Endo
Takeshi
T
Fujii
Junichi
J
Taniguchi
Naoyuki
N
D000818:Animals; D000831:Animals, Newborn; D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004274:DNA, Recombinant; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D005978:Glutathione / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D005980:Glutathione Reductase / Q000235:genetics* / Q000378:metabolism; D007668:Kidney / Q000201:enzymology* / Q000254:growth & development*; D008168:Lung / Q000201:enzymology* / Q000254:growth & development*; D016297:Mutagenesis, Site-Directed; D018384:Oxidative Stress; D051381:Rats; D017208:Rats, Wistar; D018411:Spodoptera / Q000378:metabolism; D014162:Transfection; D000094765:Glutathione Peroxidase GPX1
D018076:DNA, Complementary; D004274:DNA, Recombinant; D005979:Glutathione Peroxidase; D005980:Glutathione Reductase; D005978:Glutathione; D000094765:Glutathione Peroxidase GPX1