Metabolic deviation of mouse liver by RhIL1-alpha or RhTNF/cachectin.

In this work deviation of liver metabolism by cytokines, especially recombinant human interleukin 1-alpha (rhIL1-alpha), was investigated. Administration of rhIL1-alpha or recombinant human tumor necrosis factor (rhTNF/cachectin) to normal mice resulted in rapid, dose-dependent induction of high liver ornithine decarboxylase (ODC) activity. The effects of these cytokines on liver ODC were not indirect effects mediated by eicosanoids. The induction of liver ODC by rhIL1-alpha was at least partly a direct effect on hepatocytes, and was due to increase in de novo synthesis of the enzyme protein after increase in ODC mRNA. No specific protein was required for increase in the level of ODC-mRNA. On IL1 treatment, actinomycin D caused superinduction of liver ODC, which was at least partly due to increased stability of the ODC enzyme, because actinomycin D doubled the apparent half-life (from 50 to 95 min). Daily administration of 2 x 10(3) U of rhIL1-alpha to mice for 3 days also caused decrease in the level of the differentiated type of pyruvate kinase isozyme (PK-L) and marked increase in that of the prototype isozyme (PK-M2) in the liver, but did not cause significant change in the isozyme patterns of the kidney, thymus, and spleen. RhIL1-alpha also induced hypertrophy of the spleen. These results indicate that rhIL1-alpha causes metabolic deviation of the liver similar to that in tumor-bearing hosts.     

The in vivo stimulation of rat liver ornithine decarboxylase activity by dibutyryl cyclic adenosine 3',5'-monophosphate, theophylline and dexamethasone

The hepatic ornithine decarboxylase (ODC) activity of normal rats was stimulated more than 7-fold 3 hours after a single intraperitoneal injection of dibutyryl cyclic adenosine 3′,5′-monophosphate (dibu-cAMP). The 3-hour ODC activity was also stimulated by single injections of either theophylline or dexamethasone (10- and 21-fold, respectively). The simultaneous administration of actinomycin D with either dibu-cAMP, theophylline or dexamethasone reduced the 3-hour ODC activity by 91, 62 and 58 percent, respectively. When actinomycin D was given one hour after dibu-cAMP, no inhibition of ODC activity was observed.     

Induction of Brain Ornithine Decarboxylase During Recovery from Metabolic, Mechanical, Thermal, or Chemical Injury

Metabolic, mechanical, thermal, and chemical injury induced ornithine decarboxylase (ODC) activity in rat brain. A two- to sixfold increase in ODC activity was measured at 5-9 h after different modes of injury to the brain. During the early phase of recovery from transient ischemia, when average protein synthesis was less than 50% of control, ODC activity was increased nearly fivefold. The rise in activity could be blocked by anisomycin, or reduced by intracerebral injections of actinomycin D. Drilling burr holes into the skull, injection of the vehicle for actinomycin D, hyperthermia, and freezing lesions all caused increased ODC activity. Neurotoxic chemicals (ammonia, methionine sulfoximine, acrylamide, carbon tetrachloride, and anisomycin) also increased brain ODC activity, whereas other chemicals (mannitol and valine) did not. Treatments known to stimulate the synthesis of heat shock proteins (carotid occlusion, hyperthermia, Cd2+, canavanine, and ethanol) induced ODC activity in the liver, whereas only hyperthermia and ethanol caused significant increases in spleen ODC activity. All increases in ODC activity were blocked by difluoromethylornithine, an irreversible inhibitor of ODC. The cellular response to noxious or stressful stimuli includes the synthesis of a small number of proteins of unknown functions; ODC may be one of these "heat shock" or "trauma" proteins.     

Developmental regulation of ornithine decarboxylase (ODC) in rat testis: comparison of changes in ODC activity with changes in ODC mRNA levels during testicular maturation.

These studies were undertaken to analyze the changes in testicular ornithine decarboxylase (ODC) mRNA levels and ODC activity in rats from birth to maturity. Levels of ODC mRNA were initially low in animals aged 10-17 days. Beginning at 21 days, ODC mRNA levels began to rise, reaching maximal levels by 40 days (p less than 0.01). The size of the 2.2- and 2.6-kb ODC mRNAs did not appear to change with age, as determined by Northern blot analysis. The increase in ODC mRNA that began at 21 days paralleled the increase in testis weight. This increase in ODC mRNA preceded the appearance of rat protamine-1 mRNA, a germ cell-specific mRNA found in round spermatids, which was first detected on Day 40. In contrast, levels of sulfated glycoprotein-2 mRNA, which, in the testis, is found exclusively in Sertoli cells, were highest at Day 17 and thereafter declined gradually with age. Unlike the increase in ODC mRNA levels, ODC activity was highest in 10-day-old animals and thereafter declined steadily with age, reaching minimal levels by 40 days (p less than 0.01). Thus, the increase in testicular ODC mRNA levels was in marked contrast to the decrease in testicular ODC activity. Incubation of cytosolic extract from 40-day-old animals with that from 10- or 17-day-old animals inhibited ODC activity approximately 50%, when compared to cytosols from 10- or 17-day-old animals. Dialysis of cytosol from 40-day-old animals prior to incubation with cytosol from 10-day-old animals relieved this inhibition.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ornithine decarboxylase induction and polyamine levels in the kidney of estradiol-treated castrated male rats

Ornithine decarboxylase (ODC), S-adenosylmethionine decarboxylase (SAMDC), and thymidine kinase (TK) activities and polyamine concentrations on the kidneys of male castrated rats were studied following sc injection of estradiol. Estradiol caused an 11-fold increase in ODC activity 24 hours after administration. SAMDC activity doubled but TK activity decreased by two-thirds 2 days after estradiol treatment. The concentrations of polyamines, especially putrescine, showed sharp elevations 2 days following estradiol treatment, 1 day after the peak of ODC activity. The increase in ODC activity was suppressed by cycloheximide and by actinomycin D. Estradiol and diethylstilbestrol (DES), but not progesterone increased ODC activity. Estradiol suppressed ODC activities of liver, thymus, adrenal glands, testes and prostate. A specific estradiol-binding protein was demonstrated in the rat kidney. The dissociation constant (Kd) was 1.64 × 10^?10 M and numbers of binding sites were 31 fmoles/mg protein. Correlation between the binding of estradiol to the cytosol protein and elevation of ODC by estradiol was observed.     

Ornithine decarboxylase induction and the cytoskeleton in normal and transformed cells

ODC induction by fresh medium added to stationary, medium-depleted, confluent cultures has been studied in transformed HeLa and CHO cells, and in normal human fibroblasts as an indicator of the resumption of cell multiplication. The transformed HeLa cell displays a more easily reversed G₁ block, a higher peak ODC level, and a shorter time period for achievement of the peak ODC value than does the normal fibroblast. Low concentrations of microtubule depolymerizing agents like colchicine suppress ODC induction almost completely in the normal fibroblast, but hardly at all in the HeLa or CHO cells. Both transformed cells occasionally reveal a superinduction of ODC at very low colchicine levels (10^?8-10^?7 M) and a more variable response to such agents than does the normal fibroblast. Higher concentrations of colchicine suppress ODC induction in all cells. Experiments with actinomycin D and cycloheximide indicate that the principal colchicine action involves inhibition at the level of protein or mRNA synthesis, rather than inactivation of the already synthesized enzyme. These experiments are provisionally interpreted as an indication that a microtubular system is needed to reinitiate certain steps associated with growth in G₁-blocked, normal cells, and that a second microtubular action terminating enzyme biosynthesis may exist. This microtubular control is defective in the transformed cells here studied. Specific microtubular actions necessary for initiation and termination of protein syntheses may occur throughout the cell reproductive cycle, and in the course of normal differentiation processes.     

Cyclic AMP-dependent regulation of ornithine decarboxylase activity in Chinese hamster ovary cells maintained with a salts/glucose medium.

Ornithine decarboxylase activity (ODC) increased about 7 fold 6--8 h following 10mM asparagine (ASN) addition to confluent cultures that had been previously serum deprived and then placed in a salts/glucose medium. Optimal concentrations of dibutyryl cAMP (dB cAMP) when incubated with the ASN caused up to a 50 fold increase in the activity of this enzyme after 7--8 h. The enhancement of ODC activity by ASN and dB cAMP was not sensitive to continuous (0--7 h) treatment with actinomycin D but similar treatment with cycloheximide depressed enzyme activity 40--60%. The synergistic stimulation of ODC activity by dB cAMP added with ASN was dose dependent and the dB cAMP stimulation of ODC activity displayed an absolute requirement for ASN when cells were maintained in the salts/glucose medium. The addition of dB cAMP always further enhanced ODC activity above the levels produced by addition of various levels of ASN (1 to 40mM) to the salts/glucose medium. Other agents which elevated cAMP levels such as 1-methyl-3-isobutylxanthine (IBMX) also enhanced ODC activity when administered with ASN. Additionally, treatment with sodium butyrate at concentrations ranging from 0.001mM to 5.0mM did not elevate ODC activity above the activity obtained with ASN alone. Addition of dB cAMP at various times after placing cells in salts/glucose medium with ASN further stimulated ODC activity only when added during the first 3-4 h. These results demonstrate the involvement of cAMP in the ASN mediated stimulation of ODC activity using cells maintained in a salts/glucose medium.     

Differential effects of sex steroids on uterine and renal ODC activity in ovariectomized rats

Many hormones are known to induce the activity of ornithine decarboxylase (ODC), the first and rate-limiting enzyme for polyamine biosynthesis, in their target tissues. Using ovariectomized rats, we have compared the effects of sex hormones on ODC activity in the uterus and the kidney which contain estrogen and androgen receptors. The results show that 1) both estrogen and androgen stimulate renal ODC activity, 2) estrogen but not androgen effectively increases ODC activity in the uterus, 3) estrogen at higher dosage can stimulate renal ODC activity to an extent similar to that in the uterus, 4) daily treatment with estradiol for 5 days results in the desensitization of uterine ODC activity, but not that of renal ODC activity to the hormonal stimuli. Although both uterus and kidney are targets of sex hormones, our results indicate that estrogen and androgen have differential influences on the ODC activity in these two organs.     

Ornithine decarboxylase activity in foetal rat brain cells and in the mouse embryo fibroblasts C3H/10T1/2 CL8 cells: differences in response to medium change and to the tumor promoter TPA

12-O-tetradecanoyl-phorbol-13-acetate (TPA) induced ornithine decarboxylase (ODC, EC 4.1.1.17) in normal, preneoplastic and malignant rat brain cells in culture, but treatment with phorbol, acetate or medium shift resulted in a similar response. Medium shift induced ODC activity in C3H/10T1/2 CL8 cells 4 and 12 hr after treatment. TPA induced only the 12 hr peak. ODC induction in C3H/10T1/2 CL8 cells was completely inhibited by cycloheximide and actinomycin D. Addition of alpha-amanitin abolished the 12 hr peak, but the TPA induced ODC activity was only partly inhibited. ODC induction by TPA was lower in C3H/10T1/2 CL8 cells initiated with 3-methyl-cholanthrene (MCA). ODC increased with TPA up to 10(-7) M and decreased at higher concentrations of TPA.     

The role of messenger RNA in tyrosine aminotransferase superinduction: effects of camptothecin on hepatoma cells in culture

Camptothecin inhibited the hydrocortisone but not the insulin induction of tyrosine aminotransferase activity in hepatoma cells in culture. However, camptothecin did not cause “superinduction” of tyrosine aminotransferase activity even though it reportedly inhibits messenger RNA synthesis. In hydrocortisone pre-induced cultures, camptothecin treatment caused a rapid decline in tyrosine aminotransferase activity suggesting it did not block degradation of the enzyme. A comparison of actinomycin D with camptothecin indicated that some of the effects of actinomycin D on tyrosine aminotransferase activity may not be mediated through inhibition of messenger RNA synthesis.     

Induction of ornithine decarboxylase in cultured mouse L cells. I. Effects of cellular growth, hormones, and actinomycin D.

The activity of ornithine decarboxylase [EC 4.1.1.17] (ODC) in mouse L cells in the confluent state was induced within 4 hr by cyclic AMP (cAMP) or by insulin. During growth of L cells the concentration of cAMP increased first, then induction of ODC occurred and finally the cell number increased: the levels of cAMP and ODC increased only transitorily and returned to the basal levels when the cells become confluent. In growing cultures, however, the presence of cAMP reduced induction of ODC and cell growth. These results suggest that cAMP is involved in induction of ODC and that its concentration may be important for enzyme induction as well as for cell growth. Actinomycin D with or without these inducers stimulated induction of ODC in L cells, whereas cycloheximide inhibited it, suggesting that these hormones affect the translational level of ODC synthesis. The effect of actinomycin D on induction of ODC was much greater in non-growing cells than in growing cells. It was also found that the half life of ODC was 81 min in non-growing cells and 112 min in growing cells. This suggests that turnover of the enzyme is more rapid in the non-growing than in the growing state and that there may be an RNA fraction which controls its turnover and which also has a very short half life.