Spermine and spermidine polyamines in the brain and liver of rats under hyperoxic action

The effect of hyperbaric oxygenation (6 atmospheres) on the content of polyamines spermine and spermidine in the rat brain and liver was studied. A decrease of the spermidine content in the brain and liver during oxygen convulsions and four hours after decompression was revealed. The spermine content in the brain during oxygen convulsions decreased as well, but four hours after decompression it increased significantly not eaching the control level. The spermine content in the liver failed to alter during oxygen convulsions, but the next four hours rose sharply. The role of polyamines in the regulation of protein biosynthetic processes under hyperoxia is discussed.     

Inhibition of Na,K-ATPase from chick brain by polyamines.

The amounts of the polyamines putrescine, spermine and spermidine as well as the Na,K-ATPase activity have been determined in the developing chick brain. The amounts of spermine and spermidine per gram fresh weight do not change significantly, the amount of putrescine declines until the 17th day of incubation after which an increase takes place. Spermine is able to inhibit the Na,K-ATPase from chick brain competitively. Half maximal inhibition is achieved at 4 X 10(-5) mol/1 spermine. This polyamine functions as an allosteric inhibitor; the Hill coefficient is 2.2 +/- 0.3. A regulatory effect of spermine on the Na,K-ATPase from chick brain is discussed. In contrast to spermine 1 mmol/1 spermidine inhibits the Na,K-ATPase only slightly, while 1 mmol/1 putrescine does not inhibit the Na,K-ATPase at all.     

Selenomethionine induces polyamine biosynthesis in regenerating rat liver tissue

Summary. Our study was undertaken to elucidate the effects of selenomethionine (SeMet) on polyamine metabolism in regenerating rat liver tissue, as useful model of rapidly growing normal tissue. We have examined the levels of spermine, spermidine and putrescine in liver tissue. At the same time we have evaluated the activities of polyamine oxidase (PAO) and diamine oxidase (DAO), the catabolic enzymes of polyamine metabolism. The obtained results suggest that polyamine levels in regenerating liver tissue, at 7^th day after two-thirds partial hepatectomy, were higher in comparison with control group. The administration of selenomethionine to hepatectomized animals during seven days, in a single daily dose of 2.5 μg/100 g body weight, increases the amount of spermine and spermidine; the level of putrescine does not change under the influence of SeMet in regenerating rat liver tissue. PAO activity is lower in regenerating hepatic tissue than in control group. Supplementation of hepatectomized animals with SeMet significantly decreases the activity of this enzyme. DAO activity was significantly higher in hepatectomized and in operated animals treated with SeMet compared to the sham-operated and control ones. The differential sensitivity observed in our model of highly proliferating normal tissue to SeMet, compared with the reported anticancer activity of this molecule is discussed.     

Evaluation of Polyamine and Proline Levels during Low Temperature Acclimation of Citrus

The polyamines (PA) putrescine (Put), spermidine (Spd), and spermine (Spm) were measured during 3 weeks exposure to cold hardening (15.6°C day and 4.4°C night) and nonhardening (32.2°C day and 21.1°C night) temperature regimes in three citrus cultivars: sour orange (SO) (Citrus aurantium L.), `valencia' (VAL) (Citrus sinensis L. Osbeck), and rough lemon (RL) (Citrus jambhiri Lush). The changes in PA were compared to the amount of free proline, percent wood kill and percent leaf kill. A 2- to 3-fold increase in Spd concentrations were observed in hardened RL, SO, and VAL leaves compared to nonhardened leaves. Spermidine reached its highest level of approximately 200 nanomoles per gram fresh weight after 1 week of acclimation in both SO and VAL leaves, while RL spermidine content continued to increase up to the third week of acclimation. Spm levels in acclimated VAL and RL leaves increased 1- to 4-fold. However, SO leaves Spm content decreased with acclimation. Putrescine levels in SO and VAL increased 20 to 60% during the first 2 weeks of acclimation then declined after 3 weeks. RL putrescine content was not affected by cold acclimation. The data presented here provided direct relationship between increased Spd concentration and citrus cold hardiness. Free proline was 3- to 6-fold higher in acclimated than in nonacclimated trees. Results also demonstrate that in acclimated versus nonacclimated citrus trees the absolute amount rather than the ratio of increase in free proline is more important in predicting their ability to survive freezing stress.     

S-Adenosylmethionine Decarboxylase Levels in Rat Retina During Postnatal Development

Abstract: S -Adenosylmethionine decarboxylase from rat retina is similar to that isolated from other rat tissues with regard to kinetic parameters. pH optimum, putrescine requirement, and sensitivity to spermine. The enzymic activity increases during the first 7 days of postnatal life but decreases until the 20th day. After this period AdoMet decarboxylase activity increases, to reach the highest values at the 90th day. This behavior suggests that such enzymic activity is responsible for spermidine and spermine levels in rat retina and that a high content of retinal spermine might have a role in the photoreceptor outer segment renewal.     

Contents of putrescine, spermidine and spermine in tissue of the human brain glial tumors

It is shown that in the tissue of the human brain glial tumours the content of putrescine depends on the degree of the tumour malignization. In malignant gliomas (glioblastomas), as compared to the benign (astrocytomas), the content of putrescine is significantly higher. The content of spermidine in glial tumours of a malignancy different degree is twice as high as the level of this polyamine in the brain grey matter, and it is twice as low as in the white matter. The content of spermine in the brain glial tumours does not differ essentially from its level in the brain tissue.     

Participation of polyamines in the flowering of the short-day plant Pharbitis nil

The effect of the exogenous application of polyamines on the flowering induction of the short-day plant Pharbtis nil was investigated. Putrescine, spermidine and spermine applied on the cotyledons of 4-day seedlings had no significant effect on the flowering of this plant under conditions of full induction caused by a 16-hour-long inductive night. Under the conditions of partial induction caused by a 13-hour-long subinductive night, polyamines inhibit or stimulate flowering, depending on the time of application. Also, inhibitors of the biosynthesis of polyamines influenced the flowering process. Analysis of endogenous polyamines revealed significant fluctuations in their content in cotyledons during an inductive night, as well as under continuous light conditions. Particularly large changes occurred in spermidine and spermine levels. The putrescine level in induced seedlings was lower than in non-induced ones. However, induced seedlings contained a higher level of spermine and spermidine. The highest spermidine and spermine levels were observed at the 8th h of the night, although the total concentration of spermine during photoinduction was always 2–3 times lower than that of spermidine. A break in the inductive night, leading to a complete inhibition of flowering, had caused significant changes in the polyamine level by the end of the night. The results suggest that the flowering induction of Pharbitis nil took place at a low putrescine level and increased spermidine and spermine levels.     

The polyamine spermine induces cystocarp development in the seaweed <Emphasis Type="Italic">Grateloupia</Emphasis> (Rhodophyta)

Polyamines such as putrescine, spermidine and spermine are ubiquitous aliphatic amines involved in reproductive events in plants and algae, and first become evident through changes in endogenous levels during reproductive development. To examine whether the differences observed in polyamines, during carposporogenesis, in the red alga Grateloupia, followed a specific pattern as is seen in other organisms, infertile axes (i.e. not showing cystocarps) were excised from the same holdfast of female fertilized individuals (i.e. showing cystocarps in other axes), and cultivated until the cystocarps became visible. Changes in the endogenous levels of free putrescine, spermidine and spermine were monitored over the 8 days of culture. The activity of enzymes related to polyamine metabolism, such as l-ornithine decarboxylase (ODC), diamine oxidase and polyamine oxidase, was measured at the beginning and end of the experimental period. Up to 50% of the infertile axes became fertile and produced cystocarps at a density of 1.91 ± 0.1 cystocarps mm⁻² after 8 days. The endogenous content of spermine increased markedly over the first 5 days of culture, then decreased to the initial level by day 8. Spermidine followed a similar pattern to spermine, whereas putrescine remained at high levels, until day 5 when it decreased abruptly. The activity of ODC was less on day 8 than on day 0, whereas the activities of diamine oxidase and polyamine oxidase increased. In parallel experiments with explants from infertile axes, exogenously added spermine (10⁻⁶ M) increased the number of cystocarps, and reversed the effect of cyclohexylamine (CHA), which is known to inhibit polyamine synthesis in Grateloupia. Serial sectioning and microscopic observation of specimens from explants cultivated in 10⁻⁶ M spermine indicated that cystocarp development was induced. The results suggest that, during transition from infertile to fertile spermine is accumulated, thus favouring the development of cystocarps, given the presumed role of spermine as an inducing agent.     

Effect of the acclimation to high environmental temperature on the activity of hepatic glycogen phosphorylase (a+b and a), liver glycogen content and blood glucose level in rats

(1) Changes in the activity of hepatic glycogen phosphorylase a+b and a (GPh-ase a+b and a), liver glycogen content and blood glucose level during acclimation to moderate high environmental temperature (35±1 °C) were studied. (2) Experiments were carried out on adult fed Wistar rats of both sexes, previously given either short-term (1, 4 and 7 days) or long-term (14, 21, 30 and 60 days) exposure to high environmental temperature. The controls were continuously kept at room temperature (20±2 °C). (3) The results obtained showed that in the period of short-term exposure the liver glycogen content was decreased significantly (after the first and fourth days in male rats and after first day in female rats) and the GPh-ase a activity increased (after first day in male rats and after first, fourth and seventh day in female rats). Long-term exposure caused significant increased liver glycogen content (beginning from the 14th day in male rats and the 21st day in female rats) until the end of the acclimation period (60 days). The elevated activity of GPh-ase a persists after 14th day of exposure only in female rats while there are no significant changes over the rest of the acclimation period in both sexes. There were no significant changes in total GPh-ase activity during the whole period of exposure. Blood glucose level was significantly decreased throughout the whole period of acclimation to high environmental temperature, in both sexes (except in the 1 day exposed groups). (4) The increased activity of hepatic GPh-ase a and decreased glycogen content suggested that the short-term exposure to heat stimulates the glycogenolytical processes. Decreased blood glucose level, and elevated liver glycogen content (r=-0.7467 in male and r=-0.6548 in female rats) suggested that prolonged exposure to high environmental temperature stimulated glycogenogenesis, without changes in the GPh-ase activity.     

THE DISTRIBUTION AND METABOLISM OF PUTRESCINE, SPERMIDINE AND SPERMINE INJECTED INTO THE CEREBRAL VENTRICLES OF RABBITS

Following the intracerebroventricular injection into rabbits of spermidine or spermine the highest concentrations were initially found in the caudate nucleus, hypothalamus and medulla. Subsequently there was a rapid decline in the amounts present in the caudate nucleus and hypothalamus and, particularly in the case of spermidine, an increase in the conccntration in the lower brain stem and cervical cord. This pattern of changes is consistent with the amines being redistributed by passage in CSF. Intraventricularly injected putrescine followed the same initial distribution pattern but within 2 days it had been largely converted to spermidine and spermine. Synthesized polyamines accumulated in all the regions examined. The time course of synthesis indicated that spermidine was the precursor of spermine. Spermine was also formed from injected spermidine and vice-versa. These findings concur with the pharmacological and neurotoxic actions of putrescine, spermidine and spermine.     

Ornithine decarboxylase, transglutaminase, diamine oxidase and total diamines and polyamines in maternal liver and kidney throughout rat pregnancy.

Ornithine decarboxylase (ODC; EC 4.1.1.17), transglutaminase (EC 2.3.2.13), diamine oxidase (DAO; EC 1.4.3.6) and total di- and poly-amines were studied in rat liver and kidney cortex throughout pregnancy. In liver, ODC activity exhibited two major peaks (4.5-5 times the control activities) on days 15 and 17. Also putrescine and spermidine increased biphasically (3-4-fold), but no variation in spermine content was observed. Transglutaminase activity showed slight variations only near the end of gestation. In kidney, ODC activity did not fluctuate significantly during pregnancy, whereas both transglutaminase activity and putrescine content showed three major increases, in very early, middle and late pregnancy. No significant variations in spermidine and spermine were observed. In both organs, DAO activity, very low or undetectable until day 10, dramatically increased (10- and 20-fold in kidney and liver respectively) in the second half of pregnancy, reaching maxima on days 16-17 and 19. The results obtained for transglutaminase, ODC and total di- and poly-amines are interpreted on the basis of hyperplastic and hypertrophic events in the liver and kidney respectively. The behaviour of DAO suggests that the enzyme plays an important role in the control of intracellular diamine concentration.     

Changes in the nuclear polyamine content of chick erythrocytes during embryonic development.

The polyamine content of the circulating erythrocyte population in the embryonic chick was studied during its development. Total cellular polyamine content fell dramatically between 5 and 7 days of development, paralleling the decrease in metabolic activity exhibited by these cells. Nuclei were isolated from the erythrocytes by a non-aqueous technique, which not only eliminated the polyamine loss that occurred with aqueous isolation, but also prevented redistribution of the polyamines from the cytoplasm. Nuclear spermidine and spermine contents decreased markedly between 5 and 6 days of development from 31 to 10 pmol/microgram of DNA and from 33 to 18 pmol/microgram of DNA respectively. Thereafter the spermine content remained constant, but the spermidine content continued to decline. Good correlations between spermidine and RNA contents were observed in both cells and nuclei, and similarly between spermine and RNA contents in cells, but no such correlation was observed between spermine and RNA in nuclei.     

POLYAMINES and NUCLEIC ACID METABOLISM DURING DEVELOPMENT OF CHICK EMBRYO BRAIN

Abstract— Spermine and spermidine reach maximum concentrations in the chick embryo brain between the 12th and 14th day of incubation. Sucrose-density-gradient analysis of polyribosome distribution in the developing chick embryo brain, showed the presence of polyribosomal aggregates in the regions of 147 S and 206 S between the sixth and eighth day of incubation. After the 16th day of incubation the presence of heavier polyribosomal aggregates in the region of 259 S and 280 S was found. The injection of spermine or spermidine into the air space of embryos on the tenth day of incubation leads to a remarkable increase in the incorporation rate of [³H]formate into the ribosomes. Studies under similar experimental conditions, showed an increased radioactivity in the region of 147 S, 206 S, 259 S and 280 S in embryos injected with spermine or spermidine.     

Polyamine biosynthesis in arginine-starved and refed rats

Growth of rats fed with a synthetic diet was studied under control conditions (arginine-rich), arginine starvation, and arginine starvation/refeeding. Hepatic polyamine concentrations and ornithine decarboxylase (ODC-)activity were determined for each population. In the livers of arginine-starved rats putrescine was decreased to half the control content within 8 days; upon refeeding, it returned to control levels within another 8 days. Spermidine content in liver tissue of arginine-starved rats remained rather stable for 7 days, but thereafter dropped to half the original value within two days. Refeeding for a period of 11 days was not enough to restore the spermidine content. The effects of arginine starvation/refeeding on spermine were very similar to those of spermidine. ODC specific activity, when correlated with growth, was higher in livers of arginine-starved rats than in control animals. Refeeding caused a decrease in ODC-activity although growth arrest was completely released. This apparent uncoupling of growth and ODC stimulation supports the theory that ODC in rat liver is regulated at three levels: first the growth-related component which is observed after stimulation by growth-hormone; second the known feed back control by polyamines, e.g. via antizyme; third the regulation at the level of the substrate supply which has been shown in this work. This is not a unique finding since very similar results have been obtained in previous experiments with the protozoan Tetrahymena thermophila. A remarkable observation of these assays was that L-ornithine, when added to the arginine-free diet was not able to substitute for L-arginine in directing growth and growth related processes.     

Polyamine distribution and activity of their biosynthetic enzymes in the European sea bass (Dicentrarchus labrax L.) compared to the rat

1. In the liver, heart and brain of the European sea bass, putrescine concentrations are much higher than in the equivalent rat tissues; spermidine and spermine levels are smaller. 2. Ornithine decarboxylase in the bass liver is more active, but less stable than that in the rat; stability is acquired upon partial purification. Bass liver adenosylmethionine decarboxylase activity is less than that found in the rat. Both are activated and stabilized by putrescine. 4. The activating effect of putrescine decreases as the assay temperature is decreased. This may explain the high level of putrescine but low levels of spermidine and spermine in the bass liver.     

Studies of the induction of spermidine/spermine N1-acetyltransferase using a specific antiserum

A specific antiserum to rat liver spermidine/spermine N1-acetyltransferase was used to study the induction of this protein. The antiserum had no effect on the spermidine acetylating capacity of crude nuclear extracts and very little effect on the activity present in crude cytosolic extracts from control rat tissues indicating that most of this activity is not due to spermidine/spermine N1-acetyltransferase. Treatment of rats with carbon tetrachloride, spermidine, thioacetamide, or methylglyoxal bis(guanylhydrazone) produced a substantial increase in the spermidine acetylating capacity of rat liver cytosolic extracts which was exclusively due to an increase in the immunoprecipitable spermidine/spermine N1-acetyltransferase protein. Exact measurement of the extent of this increase was not possible because the basal amount was too low to determine precisely but the amount of this enzyme increased about 250-fold with 6 h of treatment with carbon tetrachloride, about 25-fold at 6 h after spermidine, about 23-fold at 24 h after thioacetamide and up to 300-fold at 24 h after methylglyoxal bis(guanylhydrazone). Treatment of rats with spermidine also increased spermidine/spermine N1-acetyltransferase in other tissues including lung, kidney, and pancreas. The spermidine/spermine N1-acetyltransferase protein was found to turn over very rapidly with a half-life of about 15 min in thioacetamide-treated rats and 180 min after carbon tetrachloride.     

THE UPTAKE OF SPERMIDINE AND SPERMINE BY SLICES OF MOUSE CEREBRAL HEMISPHERES

Abstract— Spermidine and spermine are taken up into mouse cerebral hemisphere slices by active transport and can be accumulated well above the medium concentration. The uptake process shows saturation kinetics and resembles that for amino acid uptake in that it is sensitive to temperature and inhibited by cyanide, 2,4-dinitrophenol or by the absence of glucose from the medium. However, at low initial medium concentrations spermine is taken up by a process which is insensitive to metabolic inhibitors or to temperature. It is suggested that either physical binding to a cellular constituent or exchange transport may account for this uptake. Ouabain does not inhibit polyamine uptake. Spermidine or spermine uptake is inhibited by cadaverine and putrescine. Spermine is the most potent inhibitor of spermidine uptake and vice-versa. Polyamine uptake differs from that of amino acids in that it is increased by a reduction in medium sodium or calcium content and decreased by an increase in medium potassium content. Recently taken up spermine undergoes heteroexchange with spermidine and homoexchange with recently entered spermine. Spermidine undergoes neither heteroexchange with spermine nor homoexchange.