Catecholamine content in the adrenals of rats developing under hypokinesia and muscle training

The effect of muscle training and hypokinesia on the content of catecholamines in the adrenals of rats in the postnatal ontogenesis has been studied. A systematic swimming training promotes the process of catecholamine accumulation in the adrenals, while hypokinesia slows down that process. An unitary carrying-out of maximum swimming exertion causes a sharp decrease of total content and concentration of catecholamines, at this their minimum concentration in the adrenals of developing animals does not depend on the adaptation to the locomotive regime.     

Influence of plasma substitutes on the concentration of tissue catecholamines in rats

The concentration of catecholamines was determined in the brain, heart and adrenals in normovolaemic and hypovolaemic rats after reinfusion of the lost blood and after infusion of equivalent volumes of plasma substitutes (dextran and modified gelatins). After infusion of these preparations in normovolaemic rats noradrenaline concentration increased significantly in the myocardium. It was found also that restoration of normal blood volume by infusion of plasma substitutes prevented changes in catecholamine concentrations induced by hypovolaemia in the brain but not in the adrenals.     

Catecholamine turnover in heart and adrenals of young rats: effects of rearing temperature.

1. A comparative chronological study (from birth to one month of age) was performed to establish the levels and turnover rates of catecholamines in the heart and adrenals of rats born and reared at 28 and 16 degrees C. 2. In the heart, cold induced marked increases of the norepinephrine level and turnover rate only from the second week of age. 3. In the adrenals there was considerable early stimulation of epinephrine production, but only in 28 degrees C rats. 4. The differences observed in the stimulation of the sympathetic nervous system according to the rearing temperature are possibly due to the behavior of the mother in warming the pups.     

Influence of fasting and dopamine on the tissue catecholamines diurnal variations

To define the role of catecholamines (CA) in the metabolic adaptation to fasting we examined the effect of exogenous dopamine(DA) on heat production(HP) and CA content in the interscapular brown adipose tissue(IBAT) and adrenals of control-fed and 2-day fasted rats in the morning(M) and in the evening(E). DA stimulates HP in fed rats in the M by 45% but the thermogenic effect of this CA is markedly higher in the E. However, DA had no thermogenic effect in fasted rats. The tissue CA in fed rats fluctuates diurnally: in the IBAT noradrenaline(NA) was much higher in the E while adrenaline(A) in adrenals was lower. DA in fed rats did not change the adrenal A but reduced NA content both in the adrenals and in the IBAT all over the day. Fasting depleted A from adrenals but increased NA content both in the M and in the E. Unlike the adrenals in the IBAT fasting did not affect NA content. In the adrenal gland of fasted rats DA significantly increased the A content to the equal degree during the day, while this CA had no effect on NA content of the IBAT.     

Effect of Escapable and Inescapable Stress on Levels of Catecholamines in Adrenals and Corticosterone in Blood Plasma in Young and Aged Rats

Effect of escapable and inescapable electrical stress (ES, IS) on the catecholaminergic system was studied in young (3 months) and aged (25 months) male Wistar rats on the day 3 after stress, using radioimmune analysis and high-pressure liquid chromatography. Catecholamine concentration in adrenals and corticosterone level in blood of control aged rats was lower than in young animals. On the third day after the electrical stimulation in cages with current-conducting floor, production of hormones of adrenal cortical and medullar layers rose significantly in aged rats, with a more pronounced increase of noradrenaline after IS, while of blood adrenaline and corticosterone, after ES. In young rats no significant changes in catecholamines were revealed, whereas the blood corticosterone level was increased after IS. Thus, in aged rats, a low basal level of catecholamines and corticosterone and a delayed stress response can be established. In old animals after ES, a long post-action was observed, which was quite comparable with the results obtained after IS in the both age groups.     

Strain differences in rat adrenal biosynthetic enzymes and stress-induced increases in plasma catecholamines.

We have examined in two inbred rat strains basal and stress-induced increases in plasma levels of epinephrine (EPI) and norepinephrine (NE) and compared these with activities of the adrenal enzymes involved in the synthesis of catecholamines. There were no differences in basal levels of NE and EPI in plasma of adult male rats of the Wistar-Kyoto (WKY) and Brown-Norway (B-N) strains. However, following 5 min. of intermittent footshock, plasma levels of both catecholamines were twice as high in WKY rats as in B-N rats. In the adrenals of unstressed rats, activities of tyrosine hydroxylase and dopamine-beta-hydroxylase were significantly higher in B-N rats. In addition, the adrenal weights and the contents of NE but not EPI were greater in B-N rats. Thus, in these two rat strains, the capacity of the adrenal gland to synthesize and store catecholamines appeared to be inversely related to plasma levels of NE and EPI after stress. The differences between the strains appeared to be due to differences in the rates of removal of catecholamines from the peripheral circulation as well as to differences in the rate of release of catecholamines from the sympatho-adrenal medullary system. Thus biosynthetic enzyme activities need not be related directly to the capacity to release and elevate plasma levels of catecholamines following stressful stimulation.     

The influence of adrenal medullectomy on the development of ethanol-induced cardiac hypertrophy

Cardiac hypertrophy was assessed in adrenal medullectomized and sham control rats given ethanol in a liquid diet mixture, intragastrically, in severely intoxicating doses at 8-h intervals for 48 and 96 h. The ethanol treatments produced increases of some 20% in wet and dry proportional heart weights in the control animals, but this rapid development of hypertrophy was less apparent in the medullectomized rats. Hearts of the medullectomized animals given ethanol were heavier than those of pair-matched controls given isocaloric maltose-dextrin in the diet mix. These increases were not statistically significant. The ethanol treatments produced, in addition, equivalent increases in the weight of the adrenal glands of both medullectomized and control animals. Medullectomy was evaluated by analysis of adrenal glands and urine samples for catecholamines. The majority of adrenals from the demedullated group had nondetectable amounts of catecholamines and minimal quantities were found in the remainder. Adrenaline was not detected generally in urine samples from this group. The adrenaline contents of adrenals from sham controls given ethanol were markedly reduced by the ethanol treatments, whereas the quantities in urine were many times greater than those from rats given maltose-dextrin. Urinary noradrenaline levels were increased in both control and medullectomized rats given ethanol. The results of this study identify that adrenal medullary catecholamines participate in the rapid development of cardiac hypertrophy that results from severe, continuing ethanol intoxication in the rat.     

Morphofunctional characteristics of adrenal reactivity in rats of different ages

Ultrastructural peculiarities of the cortical and medullary substance of the adrenals have been studied in mature (8-month-old) and old (26-month-old) intact white male rats and in rats of the same age groups in 3 h after electrical stimulation of the hypothalamic ventromedial nuclei. By means of certain biochemical methods contents of hormones (corticosterone, adrenaline and noradrenaline) have been estimated, as well as that of cholesterin and ascorbic acid in blood and adrenal tissues. After the hypothalamus is stimulated in the mature rats, certain peculiarities in the adrenals reactivity are revealed; they demonstrate positive ultrastructural and functional correlation of secretory cells in the cortex and medulla and are characterized with hypertrophy of the nuclei, increased amount of polyribosomes, hypertrophied Golgi complex and canaliculi of the endoplasmic reticulum and increased contents of corticosteroids and catecholamines in blood. In the old rats after stimulation of the hypothalamus, certain unequal ultrastructural and functional peculiarities are revealed. They demonstrate both disturbance of the hypothalamic regulation of the adrenal function at ageing and decreased adaptive possibilities of the adrenal secretory cells at the subcellular level.     

Induction of Adrenal Tyrosine Hydroxylase mRNA by Single Immobilization Stress Occurs Even After Splanchnic Transection and in the Presence of Cholinergic Antagonists

Abstract: Immobilization (IMO) stress elevates plasma catecholamines and increases tyrosine hydroxylase (TH) gene expression in rat adrenals. This study examined the mechanism(s) of IMO-induced changes in adrenal TH mRNA levels. Innervation of the adrenal medulla is predominantly cholinergic and splanchnicotomy as well as nicotinic receptor antagonists prevent the cold-induced rise in TH mRNA levels. In this study, the IMO-induced rise in plasma catecholamines, but not TH mRNA levels, was reduced by the antagonist chlorisondamine. Muscarinic antagonist atropine also did not prevent the IMO stress-elicited rise in TH mRNA. Furthermore, denervation of the adrenals by unilateral splanchnicotomy did not block the IMO-induced rise in TH mRNA but completely prevented the induction of neuropeptide Y mRNA. These results suggest that (1) the large increase in adrenal TH gene expression elicited by a single IMO stress is not regulated via cholinergic receptors or splanchnic innervation, and (2) there is a dissociation between regulatory mechanisms of catecholamine secretion and elevation of TH gene expression in the adrenal medulla of rats during IMO stress.     

Morphochemical study of brown adipose tissue during adrenal gland regeneration in the white rat

The interscapular brown adipose tissue in male, white Wistar rats in the 1,3,6,15,30, and 60th day of regeneration of adrenals after their enucleation weas investigated. The tissue slides were stained with H and E, with Masson's triple stain and with Sudan B. The activity of the following dehydrogenses was evaluated: Succinic dehydrogenase, delta 5,3 beta-hydroxysteroid dehydrogenase and the activity of the following enzymes; monoaminooxidase (MAO), Alkaline phosphatase (AlPh) and acid phosphatase (AcPh) was studied. Between the first ad 30th day of regeneration of adrenals morphochemical changes were observed; a great number of cells with eosinophylic cytoplasm appeared and with minute vacuols in their cytoplasm, the blood vessels became more numerous and enlarged and the activity of the enzymes tested was enhanced, instead sudanophilia was weaken and the reaction of corticosteriods significantly intensified. The changes observed may indicate on an enhanced hormonal activity. The brown adipose tissue may function as an endocrine gland which supports the regenerating adrenals of the white rat, enables survival and preservation of homeostasis during the insufficiency of the adrenal cortex after adrenal enucleation. The initiation of the endocrine activity of the brown adipose tissue in the course of regeneration of the adrenals proceeds probably without the direct participation of catecholamines originating from the adrenals.     

Hormonal and organ weight changes in rats exposed to simultaneous chronic effects of CO and NO2

Activity of the pituitary-adrenocortical, sympathico-adrenomedullary and thyroid gland systems was examined in groups of male adult rats, average weight 180 g, exposed for 2 months to a mixture of 7 mg.m-3 NO2 and 32 mg.m-3 CO in air and in groups of matched controls inhaling fresh air. Corticosterone (B) concentrations in the serum of rats were determined by the competitive protein-binding method, noradrenaline (NA) in the hypothalamus and catecholamines (CA) in suprarenals by fluorometry, thyroxine (T4) and triiodothyronine (T3) in the thyroid gland by the RIA technique. Measurements of organ weight were related to overall body weight. The data emerging from this study were evaluated using the Student t-test. Conclusions: one-month exposure led to a decrease in hypothalamic NA and an increase in rat spleen weight; increase in CA concentration in the adrenals was initially insignificant, by the end of a 2-month exposure it reached the level of statistical significance; serum concentrations of B, T3 and T4 remained unaffected and so was the weight of the body, liver, lungs, suprarenals and the hypothalamus of exposed rats; at the concentrations used, NO2 and CO acted as synergists producing a mild stressogenic reaction affecting the activity of the sympathico-adrenomedullary system of exposed rats.     

Synthesis of some S-3''-deoxyadenosyl-L-homocysteine analogues.

Condensation of 3'-deoxy-3-deazaadenosine, 3'-deoxy-7-deazaadenosine and 3'-deoxyadenosine with N,N'-bis-trifluoroacetyl-L-homocystine dimethyl ester and subsequent deprotection of the resulting N-trifluoroacetyl-S-3'-deoxyadenosyl-L-homocysteine analogues afforded S-3'-deoxy-3-deazaadenosyl-L-homocysteine, S-3'-deoxy-7-deazaadenosyl-L-homocysteine and S-3'-deoxyadenosyl-L-homocysteine respectively. 3'-Deoxy-3-deazaadenosine and 3'-deoxy-7-deazaadenosine were prepared by transformation of the corresponding ribonucleosides with 2-acetoxyisobutyryl bromide. 3'-Deoxy-7-deazaadenosine and 3'-deoxyadenosine were also converted into their 5'-chloro-3',5'-dideoxy derivatives which in turn were condensed with L-homocysteine sodium salt to give S-3'-deoxy-7-deazaadenosyl-L-homocysteine and S-3'-deoxyadenosyl-L-homocysteine which were identical with those synthesized by condensation of the protected L-homocystine with the 3'-deoxynucleosides.     

Catecholamines in the tissues and blood of rats after administration of acetaldehyde and ethanol

Acetaldehyde alone and in combination with acute and chronic ethanol intoxication has been studied for its effect on the concentration of epinephrine and norepinephrine in different brain areas, in the heart muscle, in adrenals and blood plasma of rats. Acetaldehyde is shown to enhance the epinephrine and norepinephrine levels in the brain areas which are non-specific for neuromediation of the mentioned catecholamines. The joint administration of acetaldehyde and ethanol increased the epinephrine concentration in adrenals probably due to the effect of acetaldehyde. On the contrary, the norepinephrine concentration in the heart decreased because of the action of ethanol. The authors' data show that acetaldehyde becomes an inductor of the mechanisms of hormone-mediator dissociation, thus altering the functions of vegetative-adrenal system. The results of the investigation support the hypothesis that acetaldehyde plays a significant role among pathogenic factors of ethanol intoxication, since it changes in a special way the catecholamine concentration in the brain and in peripheral tissues.     

State of the peripheral catecholaminergic systems during pharmacologic correction of immobilization stress using sodium oxybutyrate

Fluorescent microscopy and spectrofluorometry of biogenic amines were employed to study the peripheral catecholaminergic systems in immobilized rats which received sodium hydroxybutyrate. The content of catecholamines was measured in the adrenergic nerves of dura mater, vas deferens and chromaffin tissue of the adrenals. It was established that sodium hydroxybutyrate in a dose of 40 mg/kg intraperitoneally promoted returning to normal of the adrenergic mediator activity during alarm and resistance stages. The role of the peripheral catecholaminergic systems in the mechanism of the stress-protective effect of sodium hydroxybutyrate is discussed.     

Effect of the stress of unilateral adrenalectomy on the depletion of individual cholesteryl esters in the rat adrenal

Normal rats were subjected to unilateral adrenalectomy and were killed 3 hr later. The concentration and composition of the cholesteryl esters in adrenals removed at operation and after death were compared. The esterified cholesterol concentration was lower in the adrenals obtained 3 hr after surgery. Cholesteryl arachidonate decreased in concentration significantly more than any other ester, followed in order of magnitude by linoleate and oleate. The cholesteryl ester concentration of adrenals removed from sham-operated rats 3 hr after surgery was also greatly reduced. On the basis of comparison with other work on the hydrolysis of cholesteryl esters by adrenal homogenates, it is concluded that the apparent selectivity in depletion of cholesteryl esters is due to differences in their rates of hydrolysis.     

Decreased response of plasma catecholamine to stress in diabetic rats

Previously we reported that the heart norepinephrine concentration was markedly increased in diabetic rats. To further study the relationship between a disturbance in the autonomic nervous system and catecholamine metabolism in diabetes mellitus, the plasma catecholamine response to stress and catecholamine concentration of heart and adrenals were measured. Wistar male rats were made diabetic by streptozotocin and kept for 13 weeks. A silicon catheter was placed in the superior V. cava 1 week prior to the experiment. Insulin was injected subcutaneously for 3 days once daily. After an overnight fast and without anesthesia, 1 ml of blood, a control sample, was obtained and then the animals were exsanguinated. The blood was mixed with 1 mM EGTA at a final concentration and centrifuged. The tissue was homogenized with 0.4 N perchloric acid containing 1 mM EGTA and centrifuged at 10,000 x g for 20 minutes. Catecholamines were determined by high performance liquid chromatography. Normal rats responded to blood withdrawal stress, and plasma catecholamines were markedly increased, but almost no increase or an actual decrease was observed in diabetic rats. These abnormal responses were improved by insulin treatment. Heart norepinephrine was increased significantly in the diabetic rats compared with the control rats and was reduced significantly by insulin injections. Adrenal epinephrine was also significantly increased in the diabetic rats compared with the control rats, but was not significantly reduced by insulin. These result suggest a possible disturbance of catecholamine secretion in the diabetic rats.     

Effect of chronic alcohol administration on the adrenals thyroid, pancreatic islands and liver of male albino rats

In response to chronic oral ethanol administration, the adrenals, thyroid glands, pancreatic islets, and livers of male white rats show histological and chemomorphological reactions. Cytological, karyometrical, and histochemical findings from adrenal medulla indicate increased secretion and synthesis of catecholamines. Adrenal cortex shows morphokinetic changes within the meaning of progressive transformation. About 1 h after the last ethanol administration, there are no signs of inappropriate insulin secretion in the B-cells of pancreatic islets. In addition to a small lipid storage and a marked decrease of glycogen contents, the hepatocytes show karyometrical sign of an increased cell metabolism.     

Effect of preventive and regressive isosorbide 5-mononitrate treatment on catecholamine levels in plasma, platelets, adrenals, left ventricle and aorta in cyclosporin A-induced hypertensive rats

Increased vascular reactivity associated with cyclosporin A (CsA)-induced arterial hypertension might result from increased vasoconstriction and/or decreased vasodilatation. The administration of organic NO donors could have beneficial effects by the NO-cGMP reposition, but there is the risk of sympathetic nervous system worsening by neuro-hormonal counter-regulation. We evaluate the effect of preventive and regressive (curative) isosorbide 5-mononitrate (Is-5-Mn) treatment on blood pressures and on plasma, platelets, adrenals, left ventricle and aorta norepinephrine (NE) and epinephrine (E) contents, assessed by HPLC, in CsA-induced hypertensive rats. Five rat groups were tested: control (orange juice), CsA (5 mg/kg/day) and Is-5-Mn (150 mg/kg/day, bid) groups were treated for 7 weeks; preventive group (Is-5-Mn+CsA): Is-5-Mn during 2 weeks plus 7 weeks of Is-5-Mn+CsA; regressive group (CsA+Is-5-Mn): CsA during 7 weeks plus 5 weeks of CsA+Is-5-Mn. The increased BP in the CsA group was prevented, but was not reverted, by concomitant Is-5-Mn treatment. In the CsA-treated rats, there was a noticeable decrease in left ventricle NE and E contents and aorta NE levels and a moderate increase in circulating catecholamines, without significant effect in the adrenals values. When Is-5-Mn was preventively used, the CsA-induced effect on left ventricle and aorta was prevented. Concomitantly, however, the plasma-platelet catecholamine balance was disrupted, accumulating NE in plasma, whereas E increased in aorta, mimic the single Is-5-Mn-treated group. In opposition, in the group used as regressive Is-5-Mn therapy, the adrenals contents were higher compared with the CsA-group and, simultaneously, the CsA-evoked effects on circulating, left ventricle and aorta catecholamines were not reverted. In conclusion, regressive Is-5-Mn therapy was unable to attenuate CsA-induced catecholamine changes and BP values even worsened. On the contrary, preventive Is-5-Mn treatment prevented the catecholamine changes on left ventricle and aorta, but increased plasma NE and aorta E accumulation. Even though with those effects, hypertension development was totally prevented, suggesting that peripheral SNS per se cannot fully explain CsA-induced hypertension. Furthermore, Is-5-Mn might produce beneficial effects only if preventively employed but, considering the changes on peripheral catecholamine contents, a judicious evaluation of the nitrate therapy impact is recommended in order to avoid further deleterious effects.     

Effects of the S-adenosylhomocysteine hydrolase inhibitors 3-deazaadenosine and 3-deazaaristeromycin on RNA methylation and synthesis

The effects of 3-deazaaristeromycin and 3-deazaadenosine on RNA methylation and synthesis were examined in the mouse macrophage cell line, RAW264. S-Adenosylhomocysteine accumulated in cells incubated with 3-deazaaristeromycin while S-3-deazaadenosylhomocysteine was the major product in cells incubated with 3-deazaadenosine and homocysteine thiolactone. RNA methylation was inhibited to a similar extent by the accumulation of either S-adenosylhomocysteine or S-3-deazaadenosylhomocysteine, with S-adenosylhomocysteine being a slightly better inhibitor. In mRNA, the synthesis of N6-methyladenosine and N6-methyl-2'-O-methyladenosine were inhibited to the greatest extent, while the synthesis of 7-methylguanosine and 2'-O-methyl nucleosides were inhibited to a lesser extent. Incubation of cells with 100 microM 3-deazaaristeromycin or with 10 microM 3-deazaadenosine and 50 microM homocysteine thiolactone produced little inhibition of mRNA synthesis, even though mRNA methylation was inhibited. In contrast, mRNA synthesis was greatly inhibited by treatment of cells with 100 microM 3-deazaadenosine and the inhibition of synthesis was not correlated with an inhibition of methylation.