Localization and differential expression of arginase II in the kidney of male and female mice
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Arginase II (AII) has been almost exclusively studied in male mammalian kidneys. Our investigations were conducted to localize AII gene expression in the female mouse kidney, and to analyze the differential expression of AII gene at the transcriptional and translational levels in the kidneys of female and male mice. Total RNAs and soluble proteins extracted from renal zones and whole kidneys were analyzed by Northern and Western blots, respectively. Mitochondrial and cytosolic proteins were analyzed by Western blot. l-[guanidino-14C]arginine hydrolysis by AII was detected in microdissected tubules and the 14CO2 released from [14C]urea hydrolysis was quantified. The results of these experiments showed that: (1) both AII mRNA and protein were highly expressed in the deep cortex and the outer stripe of the outer medulla, (2) urea was produced mainly in the proximal straight tubules (PST), (3) the 38-kDa AII protein was more abundant in the mitochondria than the cytosol, and (4) the renal content of AII mRNA and protein was about three-fold higher in female than in male mice. In conclusion, in both genders, AII gene expression is restricted to the PST and localized into mitochondria. AII gene is differentially expressed in the kidney of female and male mice since higher levels of AII mRNA, protein and activity were observed in the kidneys of the former than those of the latter. Renal AII gene expression was gender-dependent in mice but not in rats. Finally, in the PST of females, l-arginine-derived ornithine may be a precursor for the renal production of l-glutamate and l-glutamine because high levels of AII, ornithine aminotransferase and glutamine synthetase are expressed in this nephron segment.
KeywordsDissected tubules Electron microscopy Gender Gene expression Mitochondria Mouse Proximal straight tubules Renal zones
The authors are indebted to Dr. Sidney M. Morris (University of Pittsburgh, Pittsburgh, PA), Dr. Anne-Marie Lefrançois-Martinez (Université Blaise Pascal, Aubière, France), and Dr. Catherine Godinot (UMR 5534 CNRS, CGMC, Villeurbanne, France), who kindly provided the respective pBSK- AII plasmid containing the mouse AII cDNA (full-length cDNA encoding for the murine AII type, GenBank accession AF032466 ), the rabbit anti-mouse AR antibody,and the monoclonal mouse anti α-subunit F1-ATP synthase, the polyclonal rabbit anti-core 2 and the polyclonal rabbit anti-ISP. Dr. O. Levillain is indebted to Marie Thérèse Ducluseau and Professor Jean-François Nicolas, who kindly provided access to materials for RNA extraction, Jocelyne Vial for assistance, and Bernard Marchand for his contribution in the animal house.
This work was supported in part by the “Association pour la Recherche sur le Cancer” (contract 6083), and has been presented in part as oral presentation of a poster at the 2nd European Polyamine Conference “Polyamines 2000, Biological and Clinical Perspectives” (1–4 June 2000, Rimini, Italy), and as a poster at the 10th International Congress on Nutrition and Metabolism in Renal Disease (6–8 July 2000, Lyon, France).
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