Rat brain catecholamines in the initial period of acute radiation exposure

Total X-ray irradiation (0.21 C/kg of rats induces an hour later an increase in the dopamine and norepinephrine content in the grey matter by 11 and 60%, respectively. The spontaneous release of dopamine and norepinephrine from the brain synaptosomes of the irradiated rats falls causing a decrease of the initial rate of the process. Potassium chloride depolarization of synaptosome membranes of the control rat brain increases both the maximal and initial rate of the process of neuromediators' release; in irradiated animals only the maximal rate of the process grows, the initial rate of the dopamine release being unchanged and that of norepinephrine increasing but less that in the control. After irradiation maximal rate of the by synaptosomes dopamine uptake is 2.6 times as high, while that of norepinephrine is 4.6 times as low; Km of the highly specific uptake of norepinephrine decreases.     

Change in the catecholamine balance of rat hypothalamus at late periods following local irradiation and its correction with etimizol

A local exposure (15 Gy) of chest caused a disturbance in the metabolism of adrenergic mediators of hypothalamus at later times (1-12 months) after irradiation: this was manifested by a decrease in the noradrenaline level 1 month following irradiation, and by phase changes in the dopamine content and considerable deficiency of deoxyphenylatanine throughout the entire period of observation. The treatment with etymisol normalized the catecholamine balance.     

Heat shock mRNA in mouse epidermis after UV irradiation

Total RNA from murine epidermis was extracted at different times after irradiation with erythemogenic doses of ultraviolet light (UVB or UVB/UVA) and hybridized to a DNA probe from the gene of a heat shock protein (hsp 70). An intense and transitory enrichment in RNA molecules hybridizing to the DNA probe was found between 15 and 120 min after irradiation, followed by a return to control levels over the next 70 h. Dose-response analysis indicates that 30 min after the irradiation, the relative amount of RNA hybridizing to the hsp 70 DNA probe increases with the dose (within the range explored: 0-180 mJ/cm2) up to values greater than 5 times the control.     

Long-term effects of irradiation with iron-56 particles on the nigrostriatal dopamine system

Exposure to heavy ions during a Mars mission might damage the brain, thus compromising mission success and the quality of life of returning astronauts. Several workers have suggested that the dopamine system is particularly sensitive to heavy ion radiation, but direct evidence for this notion is lacking. We examined measures of brain dopamine viability at times up to 15 months after acute exposure of rats to ⁵⁶Fe (1.2–2.4 Gy). No effects were seen in brain sections stained for tyrosine hydroxylase, the classical marker for dopamine cells and nerve terminals. Locomotion stimulated by cocaine, which directly activates the dopamine system, was reduced at 6 months but not at 12 months. Furthermore, in a visually cued lever-pressing test, reaction times, which are prolonged by dopamine system damage, were identical in irradiated and control animals. However, learning times were increased by irradiation. Our data suggest that the midbrain dopamine system is not especially sensitive to damage by ⁵⁶Fe particles at doses much higher than would be associated with travel to and from Mars.     

The dopamine system in rat brain areas in the early period following irradiation with superhigh doses]

In studying the main indices that characterize the neurochemical system of biosynthesis and degradation of a dopamine neuromediator, tyrosine hydroxylase-dopamine-monoamine oxidase, in different brain regions 5-6 min, 1 and 18 h after whole-body irradiation with high energy electrons (100 Gy) the authors have revealed a 25-40% inhibition of tyrosine hydroxylase and monoamine oxidase activity, and a 40% increase in the dopamine content of basal ganglia of the brain that control behavioural reactions of the organism. The neurochemical disturbances revealed are involved in the mechanisms of early transient incapacity after irradiation with superhigh doses.     

LEVELS OF NOREPINEPHRINE AND DOPAMINE IN MOUSE BRAIN REGIONS FOLLOWING MICROWAVE INACTIVATION-RAPID POST-MORTEM DEGRADATION OF STRIATAL DOPAMINE IN DECAPITATED ANIMALS

—The effects of 2 methods of killing on norepinephrine and dopamine in mouse brain regions were examined. One method utilized decapitation, while the other method utilized heating with microwave irradiation concentrated on the head. The norepinephrine and dopamine contents of the cerebellum, medulla-pons, midbrain, diencephalon, hippocampus, corpus striatum, and cerebral cortex were determined by methods using liquid chromatography with electrochemical detection. Dopamine content in striatum was also quantitated by the method of gas chromatography with mass fragmentography. A significantly lower value for decapitated animals, as compared to the microwave heated group, was found only for dopamine exclusively in the striatum. Activities of the enzymes tyrosine hydroxylase, DO PA decarboxylase, monoamine oxidase, and catechol-o-methyltransferase in the striatum were also examined. These enzymes were totally inactivated by the microwave heating, except catechol-o-methyltransferase which was decreased approx 80%. These results support either (1) the existence of a substantial pool of dopamine in the striatum with a very rapid turnover rate or (2) a decapitation-related release and destruction of striatal dopamine. Measurements of 3-methoxytyramine in the striatum exhibit post-mortem increases corresponding to the decreases of dopamine. Use of the rapid tissue enzyme inactivation technique suggests that in vivo levels of this O-methylated dopamine metabolite are an order of magnitude lower than the results normally obtained after killing by decapitation.     

Dose dependence of the excision of ultraviolet-induced pyrimidine dimers from nuclear deoxyribonucleic acids of haploid and diploid Saccharomyces cerevisiae.

The yield of ultraviolet-induced dimers is similar for a fixed dose in both haploid and diploid Saccharomyces cerevisiae. The excision of these photo-products from the nuclear deoxyribonucleic acids of cells of both ploidies after ultraviolet incident doses of 2 times 10-3 to 4 times 10-3 ergs/mm2 decreased with the corresponding increasing dose. Postirradiation incubation in saline followed by a further incubation in nutrient medium increases the excision as compared to that seen in either nutrient medium or saline alone. Previous data regarding both pyrimidine dimer removal and the survival of haploid and diploid cells after ultraviolet irradiation and either immediate or delayed plating are discussed.     

Effect of salt solutions on radiosensitivity of mammalian cells. III. Treatment with hypertonic solutions.

V79 Chinese hamster cells were treated with hypertonic solutions of NaCl or KCl and irradiated rat various times before, during, or after exposure to the solution. In solutions of molarities between 0-2 and 0-5 M, the cellular radiosensitivity increases with the molarity of the bathing solution. At these molarities, the hypertonic solution need not be present during irradiation to sensitize cells. Furthermore, radiosensitivity of cells could be increased by exposing cells for longer times to the hypertonic solution before irradiation. At higher salt concentrations (at 1-5 to 1-8 M), significant radioprotection is observed. Survival curve data showed that this protection was characterized by an increase in DO and a decrease in n, while the survival curves of cells sensitized with 0-465 M NaCl or with lower concentrations exhibited mainly changes in DO. The 1-55 M NaCl solution must be present during radiation to give a protective effect. Prolonged exposure to the salt before irradiation reduced the amount of radioprotection afforded by the salt. The results are discussed in terms of the effects of ions on histones, cellular water structure and the cell-aging cycle.     

Influence of ionizing radiation on oxygen profiles in different types of multicellular spheroids

Human glioma (U-118 MG and U-138 MG), human colorectal adenocarcinoma (HT-29), human thyroid carcinoma (HTh 7), and hamster embryonic lung (V79-379A) spheroids were irradiated with either single doses of 16 or 40 Gy or fractionated doses of eight times 5 Gy. Oxygen profiles in the spheroids were measured with microelectrodes at different times following irradiation, and these profiles were then compared with the oxygen profiles measured in parallel cultured nonirradiated spheroids. No significant radiation-induced changes in the oxygen profiles were seen in any of the spheroids within the first few days after irradiation. The glioma spheroids did not show any significant increase in oxygen tension even after longer times; however, they were growth inhibited, and the number of S-phase cells was strongly suppressed. Increases in oxygen tension did occur in the HT-29 and V79-379A spheroids but only appeared more than a week after irradiation, when degeneration had started. Histological changes and decrease in diameter were seen in the spheroids that started to degenerate about 5 days after irradiation. Thus radiation doses in the therapeutic range did not, for the spheroids studied, produce rapid increases in the oxygen tension. When a change occurred, it appeared rather late and was probably a consequence of cell degeneration.     

The action of caffeine on X-irradiated HeLa cells. VIII. Recovery from potentially lethal damage

Recovery from potentially lethal radiation damage in HeLa S3 cells has been studied by irradiating synchronous cultures with 4 Gy at selected ages in the cell cycle, initiating treatment with 4 mM caffeine, which prevents recovery, at progressively later times up to 24-30 h after irradiation, and determining the plateau level of survival after incubation with the caffeine until 36-40 h after mitotic collection. Cell recovery appears to begin immediately after irradiation at any time during interphase: an accelerating increase in survival gives way after several hours to a linear increase which lasts for an additional several hours. The median recovery time is approximately 13 h after irradiation at any time during G1, but is markedly shorter (5-7 h) after irradiation in S or G2. The rate of recovery is slightly depressed if DNA replication is inhibited with aphidicolin after irradiation and slightly enhanced if protein synthesis is inhibited with cycloheximide. Both the rate and the extent of recovery are dependent on the location of the cells in the cycle at the time of irradiation--both functions increasing with cell age from the beginning of S, but having different age dependencies in G1. Blocking cell progression with a DNA-synthesis inhibitor before irradiation halts the age-dependent changes.     

Dopamine accumulation after dopamine beta-hydroxylase inhibition in rat heart as an index of norepinephrine turnover

Dopamine concentration in rat heart is normally very low, only a few percent of the concentration of norepinephrine. After treatment of rats with a dopamine beta-hydroxylase inhibitor, 1-cyclohexyl-2-mercapto-imidazole (CHMI), there was a rapid increase in dopamine concentration even before norepinephrine concentration had decreased perceptibility. This accumulation of dopamine was readily measured by liquid chromatography with electrochemical detection. Since the percentage change in dopamine was much greater than the percentage change in norepinephrine, especially at early times, measurement of dopamine accumulation rather than norepinephrine decline was considered as a useful measure of norepinephrine turnover. Drugs that act on noradrenergic receptors and are known to alter norepinephrine turnover were found to alter the rate of dopamine accumulation. Clonidine and guanabenz decreased dopamine accumulation after CHMI, whereas piperoxan (but not prazosin) increased dopamine accumulation after CHMI. Pergolide, a dopamine agonist whose lowering of blood pressure and cardiac rate has been suggested to be due to suppression of neurogenic release or norepinephrine, also decreased dopamine accumulation after CHMI. The results suggest that measuring dopamine accumulation may have advantages over measuring norepinephrine disappearance after dopamine beta-hydroxylase inhibition as an indicator of norepinephrine turnover in heart.     

Dopamine diffusion after microwave fixation at 986 MHz

It has been previously demonstrated by us that microwave irradiation at 2450 MHz causes diffusion of dopamine from rat brain regions of high dopamine concentration to contiguous regions low in dopamine. Because we have found that the heating pattern at 986 MHz is opposite to that seen at 2450 MHz, we attempted in the present study to eliminate DA diffusion by heating at 986 MHz. However, dopamine levels were elevated in frontal and parietal cortex, remainder of cortex, and amygdala after sacrifice by microwave irradiation at 986 MHz. Norepinephrine levels were unaffected by sacrifice method. Microwave-induced diffusion of certain substances must be considered in the interpretation of regional neurochemical data.     

Fertilizing ability in vitro of golden hamster spermatozoa after acute testicular X-irradiation]

The present study is concerned with the effect of radiation to the testis on fertilizing ability in vitro using golden hamster spermatozoa. Male hamsters at 6 and 8 weeks of age were given acute testicular X-irradiation (200 kVp, 20 mA, 0.47-0.48 Gy/min). Spermatozoa were collected from the cauda epididymides at different times after irradiation and then they were suspended in fertilization medium. After preincubation for 4-5 hr, the spermatozoa were cultured with the eggs collected from mature hamsters treated with PMSG-hCG. Fertilized eggs were examined for incidence of sperm penetration and formation of pronuclei at 4-5 hr after insemination. The fertilization rate (47.7%) at the 6th week after irradiation with a dose of 2 Gy was much lower in comparison with the control value (92.6%). However, the fertilization rates at the 3rd and 9th weeks after irradiation were 97.7 and 90.6%, respectively. In these period, no difference was found between the irradiated groups and the control groups. From the changes in sperm concentration after irradiation with a dose of 2 Gy, it was found that the fertilization rate was the lowest at the 6th week. The sensitive stage to radiation during spermatogenesis with reference to the reduction of fertilizing ability after irradiation coincides with that of decrease in the sperm concentration and sperm motility. The results of fertilization rate at the 6th week after different doses of X-irradiation (0.25-6 Gy) indicated that the reduction of fertilization rate is nearly expressed as a dose-response relationship.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in the serum fraction composition of nucleic acids in radiation injuries. Alterations in an early period after gamma-irradiation of rats

The content and fractional composition of nucleic acids of blood serum change after lethal (8 Gy) particularly superlethal (100 Gy) gamma-irradiation of rats. This concerns DNA the content of which increases by 7 times 5 h following 100 Gy irradiation. It has been shown by electrophoresis in 0.85% agarose that a heterogeneous DNA fraction with the molecular weight of (1-15) X 10(6) dalton increases. The analysis of the preparation in the polyacrylamide gel has revealed a DNA fraction that is not found in norm: the fraction possesses the electrophoretic mobility corresponding to that of nucleosome DNA.     

Late functional and biochemical changes in mouse lung after irradiation: differential effects of WR-2721

The radioprotective effect of WR-2721 on late damage after whole thorax irradiation has been studied after split doses of radiation using the standard death and breathing rate assays at monthly intervals between 3 and 15 months after irradiation, as well as two biochemical measurements of injury at 15 months, hydroxyproline (HP), an indicator of tissue fibrosis, and DNA content, an indicator of tissue cellularity. A comparison of HP/lung and breaths per minute (BPM) in each dose group in the WR-2721 and non-WR-2721-treated mice 15 months after irradiation showed that the relationship between these two assays of late lung injury was not the same. There were large dose-related increases in breathing rate corresponding to relatively small changes in HP in the lungs of mice given radiation alone. In contrast, the mice given WR-2721 before irradiation showed large dose-related increases in HP/lung, but BPM remained relatively constant independent of dose. These data suggest then that changes in breathing rate and deaths later than 9 months after whole lung irradiation may not be due to collagen accumulation in the lung. WR-2721 did protect better against late lung functional changes (protection factors (PF) = 1.6) and late deaths (PF = 1.51) than against earlier changes in these same assays (PF = 1.4 and 1.28, respectively). Although the earlier-appearing injury after whole thoracic irradiation is most likely related to lung damage with deaths and increases in breathing rate resulting from pneumonitis, the cause of the late-appearing functional injury in the lung after radiation is not clear. Thus protection of late lung damage measured from either lethality or breathing rate is not related to the prevention of lung fibrosis.     

Effects of acute central and peripheral administration of nicotine on ascending dopamine pathways in the male rat brain. Evidence for nicotine induced increases of dopamine turnover in various telencephalic dopamine nerve terminal systems

The actions of intraventicular injections and intravenous infusions of nicotine were studied on dopamine stores and turnover in discrete areas of the forebrain of normal male rats. This was done by measuring the decline of the dopamine stores after tyrosine hydroxylase inhibition using alpha-methyl-tyrosine methyl ester (H44/68). The dopamine concentrations in the various telencephalic dopamine nerve terminal systems were measured using the Falck-Hillarp methodology involving quantitative microfluorimetry. The catecholamine concentrations in the anteromedial frontal cortex were measured biochemically using high pressure liquid chromatography combined with electrochemical detection. Intraventricular experiments. The dopamine levels in discrete areas of nuc. caudatus and nuc. accumbens were significantly reduced even with the lowest dose of nicotine (1 microgram/rat). Intraventricular injections of nicotine in a dose of 100 microgram/rat produced significant increases of dopamine turnover in various types of dopamine nerve terminal systems in the nuc. caudatus, nuc. accumbens and tuberculum olfactorium, and following a dose of 10 microgram/rat increases of dopamine turnover were observed in the medial part of the nuc. caudatus. Furthermore, nicotine (100 microgram/rat) significantly increased noradrenaline but not dopamine turnover within the anterofrontal cortex. Intravenous experiments. The dopamine levels were selectively reduced by nicotine (1000 microgram/kg) in the cholecystokinin positive and negative dopamine nerve terminal systems of the nuc. accumbens. On the other hand, dopamine levels in the anteromedial frontal cortex were increased after this dose of nicotine. Intravenous infusions of nicotine (10-1000 microgram/kg) produced dose-related increases of dopamine turnover in the various dopamine nerve terminal systems analysed in the telencephalon. These effects became significant with a dose of 1000 microgram/kg/h. The dopamine terminals in the nuc. caudatus showed a higher sensitivity to intravenous infusions of nicotine, being affected by 10-100 microgram/kg of nicotine. These findings suggest that relatively low dose of nicotine via an activation of central nicotine-like cholinergic receptors can reduce dopamine concentration and increase dopamine turnover in discrete limbic and striatal areas. These actions may in part represent the neurochemical basis for the rewarding actions of nicotine and for nicotine dependence in man.     

Effect of x-irradiation on the generation of O2- in rat liver microsomes

A study was made of generation of O2- in NADPH-dependent chain of oxidation of liver microsomes in irradiated rats (7 and 10 Gy). The rate of O2- generation sharply increased at early times after irradiation. The data obtained prompt an assumption that the increase in the rate of O2- generation is perhaps connected with the changes in functioning of both NADPH-cytochrome P-450-reductase and cytochrome P-450.     

Dopamine stimulated increase of GTP levels in the rat retina

Dopamine stimulates a 7-10-fold increase of GTP concentration in whole rat retina maintained in vitro. Half-maximal stimulation of GTP levels were obtained with 10(-6) M dopamine, and significant increases in GTP levels were seen with 10(-7) M dopamine. Intracellular GTP levels were significantly increased within 4 min after exposure to dopamine and maximal effects were reached within 30 min. Dopamine agonists, apomorphine and bromocriptine, also stimulate a 7-10-fold increase in GTP concentration, whereas other catecholamines (norepinephrine, epinephrine, and isoproterenol) were less potent. Several other neurotransmitters present in rat retina (gamma-aminobutyric acid, glycine, glutamine, and taurine) had no effect on GTP levels. Although dopamine also stimulates increases in cyclic AMP levels in the retina, dibutyryl cyclic AMP and 8-bromo-cyclic AMP had no effect on GTP levels, indicating that the dopamine-stimulated increase of GTP is independent of the catalytic production of cyclic AMP by adenylate cyclase. Since dopamine-stimulated adenylate cyclase activity requires GTP, the dopamine-stimulated increase in GTP concentration described in this report may serve to facilitate dopamine stimulation of adenylate cyclase activity.     

Properties of cyclic nucleotide phosphodiesterase from plasma membranes of rat synaptosomes at early stages of acute radiation injury

Phosphodiesterase of cyclic nucleotides from membranes of rat brain synaptosomes hydrolyzes cAMP and cGMP; the maximal rate of cAMP hydrolysis is 2.5 times higher than the values of the analogous index for GMP. The enzyme is found to be activated by calmodulin. A different direction of changes in the rate of cAMP and cGMP hydrolysis is observed 1 h after total X-ray irradiation. The process of cAMP hydrolysis by phosphodiesterase is characterized by positive cooperativity which is also shown after irradiation and for the process of cGMP hydrolysis. It is established that enzyme inhibition by the reaction product takes place both in control and after irradiation.     

Pituitary function during the sexual maturation of the male rat: inhibition of prolactin secretion by exogenous dopamine

Experiments were designed to determine if the responsiveness of the anterior pituitary to the prolactin (Prl) inhibiting effects of dopamine were altered during the sexual maturation of the male rat. Initial experiments established that bolus injection of dopamine HCl into cannulated (left carotid) rats pretreated with alpha-methyl-p -tyrosine (MPT) resulted in an appropriate reduction of serum Prl and that the MPT treatment did not significantly alter testosterone or luteinizing hormone (LH). Immature (25-30 days), pubertal (50-55 days), and young adult (75-80 days) rats were studied. One hour after MPT administration a blood sample was collected, followed by administration via the cannula of 2.5, 10, 40 or 160 micrograms dopamine/100 g BW or the 5% glucose vehicle. Additional blood samples were collected 15, 30, 45 and 60 min after dopamine. MPT administration resulted in a significant increase in serum Prl compared to saline-treated controls at all ages; however, the absolute value of these increases varied significantly with age. Subsequent data were calculated in terms of the decrement in serum Prl versus Time O. In immature rats the 160-micrograms dose of dopamine resulted in a significant inhibition of Prl at 15 and 30 min when compared to glucose-treated controls. In pubertal animals, 40 micrograms dopamine was effective in inhibiting Prl at both 15 and 30 min. In young adult rats, 10 micrograms dopamine was effective at 15 and 30 min; 160 micrograms lowered Prl through 45 min. These data suggest that the responsiveness of the pituitary to the Prl inhibiting effects of dopamine increases during the sexual maturation of the male rat.