Wanda Chmurzyńska

Tissue-specific effects of testosterone on S-adenosylmethionine formation and utilization in the mouse

Exogenous administration of testosterone produced several metabolic tissue-specific changes in female mouse kidneys, but not in the liver. The hormone induced ornithine decarboxylase (ODC) activity, and also profoundly influenced metabolism of S-adenosylmethionine (AdoMet). Therefore, the activity of the AdoMet-synthesizing enzyme (AdoMet synthetase) and of cystathionine synthase, which commits homocysteine irreversibly to the transsulfuration pathway, were significantly increased. In contrast to the level of AdoMet in the liver the renal level of this metabolite was augmented, whereas the level of S-adenosylhomocysteine (AdoHcy) did not change. This resulted in an increase of the AdoMet/AdoHcy ratio. In testosterone-treated mice, pulse-labelled with [methyl-14C]methionine, the radioactivity recovered in the kidneys doubled, but in the liver remained the same. The rise in radioactivity recovered occurred mainly in TCA-soluble compounds and lipids, and to a smaller extent, in proteins and nucleic acids.

Arginine and ornithine metabolizing enzymes in testosterone-induced hypertrophic mouse kidney

Administration of testosterone to female mice causes hypertrophy of their kidneys with spectacular induction of ornithine decarboxylase and significant increase in the level of putrescine. We tried to find out whether testosterone treatment affects also the renal activities of enzymes participating in the formation and utilization of ornithine, specifically arginase and ornithine aminotransferase, and whether they are dependent on putrescine level. Swiss, CFW, DBA2 or F1 (CFW x DBA2) female and male mice were injected with testosterone (125 mg/kg) or CB 3717 (100 mg/kg). DFMO was applied in the drinking water. The activities of the enzymes were determined 24 hr or 5 days after administration of CB 3717 or testosterone, respectively. Renal activities of ornithine decarboxylase (ODC), arginase and ornithine aminotransferase (OAT) were found to be sex-differentiated. The highest activity of ODC was characteristic for the kidneys of males, whereas those of arginase and OAT for the kidneys of females. In the kidneys of testosterone-treated female mice a decrease (50%) of OAT, and a significant increase of arginase activities (up to 200%), were observed. In the males these changes were less pronounced. DFMO, which completely inhibited the activity of renal ODC, did not influence significantly the testosterone-induced arginase and the testosterone-decreased OAT. Arginase and OAT, in contrast to ODC, were not changed in CB 3717-induced hyperplastic kidney. The study showed testosterone-induced differential changes in the activity of two enzymes involved in ornithine biosynthesis and catabolism which accompanied ODC induction in female mouse kidney.(ABSTRACT TRUNCATED AT 250 WORDS)

Polyamines in testosterone-induced hypertrophic and antifolate-induced hyperplastic mouse kidney. Differential effect of α-difluoromethylornithine

In the testosterone-induced hypertrophic and antifolate (N10-propargyl,5,6-dideazafolic acid, CB 3717)-induced hyperplastic mouse kidney models, a marked increase of two diamine levels--putrescine and cadaverine--occurred which paralleled induced ornithine decarboxylase (ODC) activity. Under these conditions the augmentation of spermidine levels was much smaller, while spermine levels were affected differentially--increased by testosterone and decreased by CB 3717; this resulted in an increase of spermidine/spermine ratio in hyperplastic, but not hypertrophic kidney. alpha-Difluoromethylornithine (DFMO) prevented testosterone- or CB 3717-induced increment of both diamine levels. Spermidine and spermine depletion in response to DFMO was significant in hyperplastic kidney only. DFMO also significantly affected the other biochemical markers of hyperplasia, namely lowered CB 3717-induced cell proliferation rate and increased S-adenosylmethionine decarboxylase (AdoMetDC) activity. In contrast, testosterone-induced hypertrophy was not influenced by DFMO, as judged by the lack of its effect on S-adenosylmethionine synthetase and cystathionine and synthase activity. These results indicate that the increase of putrescine levels does not mediate testosterone-induced renal hypertrophy and possibly also antifolate-induced hyperplasia. The involvement of spermidine in mediation of renal hyperplasia is highly possible, while that of spermine is excluded.

Catecholamines are required for androgen-induced ODC expression but not for hypertrophy of mouse kidney.

Catecholamine depletion, evoked by reserpine, dramatically impaired (5-fold) the testosterone-induced increase of ornithine decarboxylase (ODC) activity in female mouse kidney. However, reserpine did not prevent kidney hypertrophy evoked by testosterone. This is evidenced by the activity of sensitive, biochemical markers of renal hypertrophy, namely arginase and ornithine aminotransferase (OAT), that responded with the increase and decrease of activities to testosterone treatment, respectively. Arginine and ornithine, substrates and/or products of marker enzymes, showed a striking homeostasis as their level was not affected by testosterone and reserpine, and only slightly by DFMO. Northern blot analysis revealed that the ODC mRNA level, that was increased 10-fold by testosterone, was decreased 2-fold in catecholamine-depleted hypertrophic kidney. Thus, ODC transcript level, lowered by reserpine, correlated partially with an attenuated response of ODC activity to testosterone. This was in contrast to DFMO, which inhibited ODC activity, but significantly increased its mRNA content. It is concluded that catecholamines could be involved together with testosterone in regulation of the ODC gene expression in mouse kidney.

Ornithine decarboxylase induction in mouse kidney as indicator of renal damage. Differential nephrotoxic effect of anticancer antifolate drugs

High doses of folate and its quinazoline analogue with antitumour activity, N10-propargyl-5,8-dideazafolic acid (CB 3717), caused severe renal damage in mice, leading in the case of folate to death. The mouse kidneys increased in weight, which was accompanied by time- and dose-dependent induction of ornithine decarboxylase (ODC) activity. In contrast, methotrexate (MTX) had negligible effect on mouse kidneys except when applied together with the non-steroidal antiinflammatory drug, indomethacin.

Age- and tumour-related changes in methionine biosynthesis in mice

Two routes for the methylation of homocysteine to methionine, depending either on betaine or folate cofactor as methyl donor, were studied in liver and kidneys of normal and Ehrlich ascites carcinoma-bearing mice at various stages of their postnatal development. Distinct age-dependence in the activities of betaine methyltransferase, methylenetetrahydrofolate reductase and methionine synthase were found both in normal and tumour-bearing mice. Independent of the levels of enzyme activity in healthy mice, the tumour activated one route of methionine formation only, namely that utilizing methyltetrahydrofolate as the methyl donor. This effect was observed in host liver exclusively. No host age-related changes were found in Ehrlich ascites carcinoma growth.

Catecholamines participate in the induction of ornithine decarboxylase gene expression in normal and hyperplastic mouse kidney

In the quinazoline antifolate (CB 3717)-induced hyperplastic kidney model, a remarkable increase of ornithine decarboxylase (ODC) activity was paralleled by a smaller, but highly significant augmentation of the ODC transcript level. Catecholamine depletion, evoked by reserpine, strongly impaired antifolate-induced ODC expression; the enzyme activity was almost completely abolished while the mRNA level decreased by 60%. Moreover, under conditions of a depleted catecholamine pool, kidney enlargement was significantly reduced confirming our earlier reports on the indispensability of ODC induction for renal hyperplasia (M. Manteuffel-Cymborowska et al. , Biochim. Biophys. Acta, 1182 (1993) 133-141[1]). In normal mouse kidney catecholamines appeared to be inducers of ODC expression. Use of selective agonists of catecholamine receptors demonstrated the importance of dopamine D2 receptors, and to a lower extent beta adrenoreceptors, in the catecholamine mediation of induction of ODC activity and of ODC mRNA levels. These increases were not abolished by an antiandrogen, casodex, suggesting that catecholamine control of ODC expression is an androgen receptor-independent process. The results obtained point to the critical role of renal catecholamines; these biogenic amines are not only involved in the regulation of ODC expression in normal kidney but are also required for the induction of ODC in hyperplastic kidney evoked by antifolate and, as shown recently (M. Manteuffel-Cymborowska et al., Biochim. Biophys. Acta, 1356 (1997) 292-298[2]), in testosterone-induced hypertrophic kidney.

Interference of a synthetic C18 juvenile hormone and related insect growth regulators with macromolecular biosynthesis in mammalian cells.

Two juvenile hormone analogs (JHA), methoprene (ZR-0515), and ZR-0619 and a synthetic C18 juvenile hormone (JH) itself were compared in their effects on protein and nucleic acid biosynthesis and on the viability of mouse L cells. A short-time exposure of the cells, suspended in phosphate-buffered saline, to the labeled precursors of protein, DNA or RNA, and to juvenile hormone, methoprene, or ZR-0619 (2–100 μg/ml) considerably depressed the synthesis of these macromolecules. The effects of the juvenile hormone were evidently stronger than those of methoprene or ZR-0619. RNA synthesis was affected to a lesser extent than protein or DNA synthesis. Under the same experimental conditions, both the JHA were less cytocidal than the juvenile hormone itself. Calf serum added to the incubation mixture protected the cells against the effects of juvenile hormone and the JHA.

Tumour effect on arginine/ornithine metabolic relationship in hypertrophic mouse kidney.

Tumour effect on arginine/ornithine metabolic relationship in hypertrophic mouse kidney. The presence of a tumour significantly changes nitrogen metabolism, including that of amino acids and polyamines, in host animals. In this study, we examine whether developing tumours affect the metabolic relationship of arginine and ornithine, precursors of polyamines, in the testosterone-induced hypertrophic mouse kidney model. Androgen-induced changes in the activity of enzymes involved with ornithine biosynthesis (arginase), its consumption (ornithine aminotransferase, OAT and ornithine decarboxylase, ODC) and the hypertrophy of host mouse kidney were not affected by the presence of an ascitic tumour (EAC) and only slightly by a mammary carcinoma (MaCa). The HPLC determined renal level of arginine and ornithine showed a striking homeostasis and was disturbed neither by testosterone nor EAC. The effect of MaCa and testosterone on the levels of both amino acids, although significant, was not very pronounced. Developing tumours, especially ascitic, altered the renal activity of OAT and ODC, but not of arginase, in testosterone-untreated mice. All examined tumours, EAC, L 1210 and MaCa actively metabolized arginine and ornithine. The tumour content of arginine which coincided with the activity of arginase, resulted in a marked increase of the ornithine/arginine ratio in tumours, when compared with kidneys. These results indicate that the androgen-induced anabolic response in mouse kidney is preserved, in spite of tumour requirements for essential metabolites.

One-carbon pathways in amino acid interconversion in the sawfly, Acantholyda nemoralis

Abstract Experiments were based on the injection of 14 C-formate or 3- 14 C-serine into diapausing last instar larvae followed by radioassays of the blood amino acids. The bulk of the radioactivity was always found in serine. Thus the radiocarbon of formate after incorporation into serine became a real source of the label in further metabolic processes. Methionine present in sawfly blood at a concentration of a few mg per cent contained some detectable radiocarbon. This is indirect evidence for the possibility of methionine biosynthesis in this insect. The probable pathways of amino acid interconversions in the sawfly are discussed.