The effect of noradrenaline on the neurogenic vasoreactivity at various frequencies of the electrical field stimulation

The effect of 0.03-10.0 microM noradrenaline on the response to electrical field stimulation (EFS) of the juvenile rat tail artery segment was studied. At frequencies of the EFS equal to 10 or 40 Hz, noradrenaline was shown to cause much more pronounced potentiation or--at higher concentration--much less pronounced inhibition of the EFS-evoked constriction in arteries characterized by spontaneous decrease in the constriction value in the course of experiments as compared with arteries which were not characterized by such a decrease. At frequencies of the EFS equal to 3 or 5 Hz, the value and/or direction of the change in the neurogenic vasoconstriction in the presence of noradrenaline depends on the presence of the spontaneous decrease in the constriction evoked by EFS at 10 Hz, rather than at 3 or 5 Hz. It is concluded that the character of the change in the neurogenic vasoreactivity is a factor of a great importance for the prediction of the further change in the reactivity in the presence of noradrenaline.     

TRANSPORT OF NORADRENALINE IN SYMPATHETIC NERVES AND THE EFFECT OF NERVE IMPULSES ON ITS CONTRIBUTION TO TRANSMITTER STORES

Abstract— —A study has been made of the contribution of noradrenaline transport along sympathetic nerves to their terminal stores of transmitter by ligating the splenic nerves of the cat, and measuring both the noradrenaline that accumulates above the constriction, and the noradrenaline content of the spleen. The biochemical estimations were supplemented by fluorescence histochemistry. The effect of abolishing efferent impulses in the splenic nerves was examined by cutting their preganglionic nerve supply.
The proximo-distal flow rate for noradrenaline was calculated as 1.4-3.3 mm/hr assuming that all the noradrenaline that accumulates is derived from the cell bodies in the ganglion without net addition or loss in the axons. The process was not dependent on impulse traffic in the nerves, since decentralization did not significantly effect the accumulation. The amount of noradrenaline arrested by the constriction in 24 hr was only 1 per cent of the stores in the terminals of those nerves, and consequently no change was detected in the spleen's noradrenaline content as a result of constricting its nerve supply.
In the presence of an intact reflex pathway to the spleen, the stress of the operative procedure produced a marked constriction of the spleen, and depletion of its noradrenaline content. These changes could be prevented either by section of the preganglionic splanchnic nerves, or by ligation of the splenic nerves, thereby blocking the conduction of efferent nerve impulses.
The evidence favours a proximo-distal flow of noradrenaline in sympathetic nerves, independent of nerve impulses, which makes, however, a negligible quantitative contribution to the terminal stores of transmitter.     

Effect of nitric oxide in protective effect of melatonin against chronic constriction sciatic nerve injury induced neuropathic pain in rats

Developing a successful treatment strategy for neuropathic pain has remained a challenge among researcher and clinicians. Various animal models have been employed to understand the pathogenic mechanism of neuropathic pain in experimental animals. The present study was designed to explore the possible nitric oxide mechanism in the protective effect of melatonin against chronic constriction injury (CCI) of sciatic nerve in rats. Following chronic constriction injury, various behavioral tests (thermal hyperalgesia, cold allodynia) and biochemical parameters (lipid peroxidation, reduced glutathione, catalase, and nitrite) were assessed in sciatic nerves. Drugs were administered for 21 consecutive days from the day of surgery. CCI significantly caused thermal hyperalgesia, cold allodynia and oxidative damage. Chronic administration of melatonin (2.5 or 5 mg/kg, ip) significantly attenuated hyperalgesia, cold allodynia and oxidative damage in sciatic nerves as compared to CCI group. Further, L-NAME (5 mg/kg) pretreatment with sub-effective dose of melatonin (2.5 mg/kg, ip) significantly potentiated melatonin's protective effect which was significant as compared to their individual effect per se. However, L-arginine (100 mg/kg) pretreatment with melatonin (2.5 mg/kg, ip) significantly reversed its protective effects. Results of the present study suggest the involvement of nitric oxide pathway in the protective effect of melatonin against CCI-induced behavioral and biochemical alterations in rats.     

Noradrenergic Inhibition and α2-Adrenergic Stimulation of Melatonin Secretion in the Pigeon

Adrenergic regulatory mechanisms of melatonin synthesis and secretion were studied in the pigeon in vivo. Late-afternoon intraperitoneal injection of noradrenaline (NA; 1 mg/kg) resulted in a significant decrease in plasma melatonin levels in 3 h. The same effect was seen after phenylephrine treatment (1 mg/kg i.p.), indicating that an alpha 1-adrenergic mechanism may mediate the inhibition. Propranolol treatment had no effect on plasma melatonin levels, supporting this concept. Detomidine (1 mg/kg i.p.), an alpha 2-adrenergic agonist, increased melatonin levels. This stimulatory effect was blocked by yohimbine, an alpha 2-adrenergic antagonist. However, yohimbine alone had no effect on the plasma melatonin levels, suggesting that alpha 2-adrenergic transmission is not primarily responsible for the nocturnal stimulation of melatonin synthesis and secretion in the pigeon.     

Effect of melatonin on neurogenic vasoreactivity: formation and modulation of the vessel response

The effect of melatonin (0.1 microM) on the contractile response to electrical field stimulation (EFS) of the juvenile rat tail artery segment was studied. The spontaneous decrease in reactivity of the segment observed in the course of our experiments was accompanied with a melatonin-evoked increase in the reactivity which was proportional to this decrease and was not connected with a sensitization of the segment to the substance. Perfusion of the segment with an acidic solution leads to a more pronounced inhibition of the response as well as to a greater melatonin-evoked protentiation of the response. Noradrenaline-evoked response was not augmented by melatonin. The results suggest that the potentiating effect of melatonin on the EFS-evoked response of the juvenile rat tail artery depends on degree of the change in the artery reactivity and was not due to change in sensitivity of postjunctional membrane to noradrenaline.     

The effect of tetrabenazine on the accumulation of noradrenaline in constricted postganglionic sympathetic nerves in vitro

The accumulation of noradrenaline proximal to a constriction applied to cat hypogastric nerves in vitro has been studied in preparations treated with tetrabenazine. The accumulation of amine was almost completely abolished by the drug. Evidence is presented which suggests that tetrabenazine exerts a direct noradrenaline-depleting effect on the intraneuronal storage vesicles. Recovery of noradrenaline levels after the removal of the drug was rapid and was effected by the biosynthesis of new noradrenaline within the axon.     

Melatonin restores diminished neurogenic reactivity of the juvenile rat tail artery

The effect of melatonin on neurogenic reactivity of the juvenile rat tail artery segment was studied. The electrical field stimulation-evoked contraction of the segment decreased in the course of the experiment. Melatonin (0.1 microM) applied at different time points of the experiment produced an increase in the contraction, which directly correlated with a spontaneous decrease in the electrical field stimulation-evoked response. The increase in the potentiating effect of melatonin in the course of the experiment was not due to sensitization of the segment to this substance. Noradrenaline-evoked contraction of the vessel segment was not changed by melatonin. The data indicate that melatonin restores the diminished neurogenic reactivity of the juvenile rat tail artery probably by potentiation of the contractile response of the vessel, but this effect is hardly due to a change in sensitivity of the postjunctional membrane to noradrenaline.     

Adrenergic mechanisms in the hepatic microcirculation in the rat

The in vivo hepatic microvascular bed of the rat was observed microscopically in the transilluminated liver and the diameter of the hepatic sinusoids was measured by serial photomicrography. Intraportal infusion of tyramine induced concentration-dependent constriction of the hepatic sinusoids, but also dilatation of the sinusoids when the dose was small. These effects were attributed to the release of endogenous noradrenaline which activated either alpha- or beta-adrenergic receptors and caused constriction, or dilatation, of the sinusoids respectively. Adrenaline and noradrenaline induced similar changes in the hepatic sinusoids as tyramine, while phenoxybenzamine induced dilatation, and propranolol constriction, of the sinusoids. All the above responses were abolished by pretreatment with reserpine. A possible noradrenaline-mediated basal vasomotor tone in the hepatic sinusoids for autonomic control of the blood flow in the sinusoids was postulated.     

Aminergic innervation pattern of the rodent pineal gland: no apparent influence of time of day

Pineal melatonin synthetic activity shows distinct diurnal characteristics. The circadian regulation of melatonin synthesis is provided by noradrenaline-releasing sympathetic nerves. The pineal noradrenaline content shows a circadian rhythmicity tidally related to the changes in melatonin synthesis rate. To evaluate possible circadian changes of pineal noradrenergic fibre arrangement, the nerve distribution in rat and guinea pig pineal glands was visualized by means of glyoxylic acid-induced histofluorescence. Histochemical findings at 08.00 h and 24.00 h did not exhibit any differences: in both species a dense, mainly perivascularly located network of fluorescent fibres was encountered. As indicated by the simultaneous intraneural presence of green-bluish and yellow fluorophores these fibres most likely contain noradrenaline and serotonin. Obviously circadian melatonin synthesis changes are not paralleled by changes in the distribution pattern of pineal sympathetic nerve fibers. Like other sympathetic innervation-related morphological parameters, histofluorescence does not accurately reflect circadian biochemical changes in the pineal gland.     

The profile of melatonin production in tumour-bearing rats

The pineal gland is involved in the regulation of tumour growth through the anticancer activity of melatonin, which presents immunomodulatory, anti-proliferative and anti-oxidant effects. In this study we measured melatonin content directly in the pineal gland, in an attempt to clarify the modulation of pineal melatonin secretory activity during tumour growth. Different groups of Walker 256 carcinosarcoma bearing rats were sacrificed at 12 different time points during 24h (12h:12h light/dark cycle) on different days during the tumour development (on the first, seventh and fourteenth day after tumour inoculation). Melatonin content in the pineal gland was determined by high-performance liquid chromatography with electrochemical detection. During tumour development the amount of melatonin secreted increased from 310.9 ng/mg of protein per day from control animals, to 918.1 ng/mg of protein per day 14 days after tumour implantation, and there were changes in the pineal production profile of melatonin. Cultured pineal glands obtained from tumour-bearing rats turned out to be less responsive to noradrenaline, suggesting the existence, in vivo, of putative factor(s) modulating pineal melatonin production. The results demonstrated that during tumour development there is a modification of pineal melatonin production daily profile, possibly contributing to cachexia, associated to changes in pineal gland response to noradrenaline stimulation.     

Suppression of sympathetic nervous system by short photoperiod and melatonin in the Syrian hamster

Exposure to short photoperiod or melatonin treatment brings about gonadal regression in Syrian hamsters. The possible influence of these treatments on the sympathetic nervous system (SNS) in these animals was investigated. Male Syrian hamsters were exposed to either long or short photoperiod or subjected to administration of melatonin or its vehicle solution. Exposure of hamsters to 10 weeks of short photoperiod, significantly reduced the noradrenaline (NA) turnover in the heart. Daily administration of melatonin for 8 weeks also resulted in a similar suppression of NA turnover in the heart. Hamsters that were treated with melatonin maintained a lowered metabolic rate as well, at and below thermoneutral temperature. These findings suggest that in a deep hibernator, short photoperiod could suppress the peripheral sympathetic activity and that melatonin may act as the endogenous mediator.     

Noradrenaline- and melatonin-mediated regulation of pigment aggregation in fish melanophores

The effects of melatonin and noradrenaline (NA) on bi-directional melanosome transport were analysed in primary cultures of melanophores from the Atlantic cod. Both agents mediated rapid melanosome aggregation, and by using receptor antagonists, melatonin was found to bind to a melatonin receptor whereas NA binds to an alpha2-adrenoceptor. It has previously been stated that melatonin-mediated melanosome aggregation in Xenopus is coupled with tyrosine phosphorylation of a so far unidentified high molecular weight protein and we show that although acting through different receptors and through somewhat different downstream signalling events, tyrosine phosphorylation is of the utmost importance for melanosome aggregation mediated by both NA and melatonin in cod melanophores. Together with cyclic adenosine 3-phosphate-fluctuations, tyrosine phosphorylation functions as a switch signal for melanosome aggregation and dispersion in these cells.     

Noradrenaline‐ and Melatonin‐Mediated Regulation of Pigment Aggregation in Fish Melanophores

The effects of melatonin and noradrenaline (NA) on bi‐directional melanosome transport were analysed in primary cultures of melanophores from the Atlantic cod. Both agents mediated rapid melanosome aggregation, and by using receptor antagonists, melatonin was found to bind to a melatonin receptor whereas NA binds to an α2‐adrenoceptor. It has previously been stated that melatonin‐mediated melanosome aggregation in Xenopus is coupled with tyrosine phosphorylation of a so far unidentified high molecular weight protein and we show that although acting through different receptors and through somewhat different downstream signalling events, tyrosine phosphorylation is of the utmost importance for melanosome aggregation mediated by both NA and melatonin in cod melanophores. Together with cyclic adenosine 3‐phosphate‐fluctuations, tyrosine phosphorylation functions as a switch signal for melanosome aggregation and dispersion in these cells.     

Effects of pressure on the skin exerted by clothing on responses of urinary catecholamines and cortisol,heart rate and nocturnal urinary melatonin in humans

The study investigated how the pressure exerted on the skin by clothing worn while working in the daytime affected the urinary excretion of adrenaline, noradrenaline and cortisol, heart rate, and also melatonin secretion at night. Nine young women (experiment I) and seven young women (experiment II) participated. Participants wore either a 100% cotton jacket (tight clothes, TC) or a 100% cotton T-shirt (loose clothes, LC). Loose-fitting, 100% cotton tank tops and panties were worn as underwear in both the TC and the LC groups. The main results can be summarized as follows: (1) urinary excretion of adrenaline, noradrenaline and cortisol was facilitated, and the amounts of urinary excretion were significantly higher when TC were worn. Heart rate was significantly higher in the TC group; (2) nocturnal urinary melatonin excretion was significantly greater in the TC group. These results are discussed in terms of an enhancement of diurnal sympathetic nervous system activity caused by pressure on the skin produced by tight clothing.     

PROXIMO-DISTAL TRANSPORT OF [14C]NOR-ADRENALINE AND PROTEIN IN SYMPATHETIC NERVES

Abstract— —Both [¹⁴C]noradrenaline and [¹⁴C]leucine were injected into the coeliac ganglia of cats in an attempt to label the noradrenaline and protein of the granular vesicles, so that their movement in the splenic nerves could be followed.
When a constriction was placed on the nerves, labelled noradrenaline and protein accumulated just proximal to it, but there was no such accumulation below it, nor above a second, more distal constriction placed on the same nerve. This indicated that a neural transport mechanism, rather than uptake from the circulation, was responsible for the accumulation.
Peaks of labelled noradrenaline and protein were observed to move down the axon at about 5 mm/hr. In addition a slow moving component of axonal protein, advancing at about 1 mm/day, was detected.
The results demonstrate a rapid proximo-distal movement of noradrenaline and protein which could represent the transport of granular synaptic vesicles from their site of manufacture in the cell body to their site of storage in the nerve terminals within the spleen.     

Evidence for different pre-and post-junctional receptors for neuropeptide Y and related peptides

The effects of neuropeptide Y (NPY), peptide YY (PYY), desamido-NPY and five C-terminal fragments of NPY or PYY were tested on different smooth muscle preparations in vitro. The fragments were NPY 19-36, NPY 24-36, PYY 13-36, PYY 24-36 and PYY 27-36. NPY and PYY appear to exert three principally different effects at the level of the sympathetic neuroeffector junction. Firstly, they have a direct post-junctional effect, leading to constriction of certain blood vessels; this was studied on the guinea-pig iliac vein. Secondly, they potentiate the response to various vasoconstrictors; this was studied on the rabbit femoral artery and vein, using noradrenaline and histamine, respectively, as agonists. Thirdly, NPY and PYY act prejunctionally in that they suppress the release of noradrenaline from sympathetic nerve endings upon stimulation; this was studied in the rat vas deferens. NPY and PYY were approximately equipotent in constricting the guinea-pig iliac vein, while desamido-NPY and the fragments were without effect. Desamido-NPY and the fragments were ineffective also in potentiating the response to noradrenaline in the rabbit femoral artery, nor did they potentiate the response to histamine in the rabbit femoral vein. NPY and PYY potentiated the response to noradrenaline in the artery, as well as the response to histamine in the vein. The NPY- and PYY-induced suppression of noradrenaline release from the prostatic portion of the rat vas deferens was reproduced by PYY 13-36 but not by the shorter fragments nor by desamido-NPY. In conclusion, a C-terminal portion seems to be sufficient for exerting the prejunctional effect of NPY and PYY, while the whole sequence seems to be required for post-junctional (direct and modulatory) effects. An amidated C-terminal is crucial for maintaining the biological activity of NPY. Desamido-NPY and the fragments that were inactive as agonists also seemed inactive as antagonists.     

beta-Adrenergic receptor subtypes in melanophores of the marine gobies Tridentiger trigonocephalus and Chasmichthys gulosus.

The subtype of beta-adrenergic receptors in melanophores of the marine gobies Tridentiger trigonocephalus and Chasmichthys gulosus was studied. Pigment of denervated melanophores in isolated, split caudal fins was preliminarily aggregated by incubating the specimens in a physiological saline containing 10 microM phentolamine and 30-100 microM verapamil or 2-10 nM melatonin, and the responses of the melanophores to a beta-adrenergic agonist added to the incubating medium were recorded photoelectrically. The beta-adrenergic agonists noradrenaline, adrenaline, isoproterenol, salbutamol and, dobutamine were all effective in evoking a dispersion of melanophore pigment in the presence of phentolamine and verapamil or melatonin. The pigment-dispersing effect of noradrenaline (beta 1-selective agonist) was inhibited by metoprolol (beta 1-selective antagonist), propranolol,- and butoxamine. Whereas, the effect of salbutamol (beta 2-selective agonist) was hardly inhibited by metoprolol, though it was considerably inhibited by propranolol and ICI-118551. It was estimated that beta 1- and beta 2-adrenergic receptors coexist at ratios of 8.6:91.4, in the melanophore of Tridentiger trigonocephalus, and 25:75, in the melanophore of Chasmichthys gulosus, through the analyses of Hofstee plots of the effects of the beta-adrenergic drugs. It was suggested that the relation between the pigment-dispersing effect of a beta-adrenergic agonist on the melanophores and the concentration of the drug follows mass action kinetics, when the effect is mainly caused by the activation of beta 2-adrenergic receptors of the melanophores. However, when it is mainly caused by the activation of beta 1-adrenergic receptors of the melanophores, the relation does not follow mass action kinetics.     

Dopamine disorganizes direct intercellular interactions in keratinocytes cultures: A comparison to hepatocytes

Dopamine in the concentration 0.4 μg/mL abolishes protein synthesis rhythm in HaCaT keratinocytes and hepatocytes unlike noradrenaline or melatonin, which synchronize direct intercellular interactions and organize protein synthesis rhythm. Experiments with D2 dopamine receptors blocking agent metoclopramide (tserukal) in the concentration 2 μg/mL show that a disorganizing effect of dopamine is driven by the activation of D2 receptors, which block adenylyl cyclase and the efflux of calcium ions from internal depos according to the literature. It is shown that tserukal does not activate serotonin receptors in our experimental settings. Cellular interactions’ recovery during or after dopamine action is carried out by melatonin in the concentration 0.001 μg/mL. A recommendation to inject melatonin before dopamine administration for different medical indications is discussed.     

Stimulation of progesterone secretion by cultured human granulosa cells with melatonin and catecholamines

Granulosa cells, aspirated from the follicles of patients undergoing treatment for in-vitro fertilization, were cultured in serum-supplemented medium. Adrenaline and noradrenaline stimulated a dose-related increase in progesterone secretion with a maximum stimulation at 10(-5) M, a response that was prevented by the beta-antagonist, propranolol. Adrenaline and hCG showed similar characteristics in their stimulation of progesterone secretion but there was no further increase in progesterone when the 2 compounds were added together. Melatonin stimulated progesterone secretion and, like adrenaline, this stimulation was prevented by propranolol. The ability of both adrenaline and melatonin to increase progesterone secretion was dependent on the degree of follicular development, as determined by peripheral oestradiol concentrations, on the day of laparoscopy. These results suggest that adrenaline and melatonin may have a physiological role in modulating luteal function and that melatonin may act by a beta-adrenergic-related mechanism.     

Melatonin potentiates the vasoconstrictive effect of noradrenaline in renal artery from newborn hooded seals (Cystophora cristata) and harp seals (Phoca groenlandica)

Isometric tension was recorded by force displacement transducers in ring segments of the inferior branch of the renal artery from newborn hooded seals (Cystophora cristata, n=6), harp seals (Phoca groenlandica, n=3) and domestic pigs (Sus scrofa f. domestica, n=5). Arterial segments were mounted in heated organ baths and exposed to graded concentrations of noradrenaline (NA) and adrenaline (A), alone or together with melatonin. The melatonin concentration in the bath was similar to the plasma concentration normally found in each experimental animal. Melatonin alone did not affect the tension in any of the species, but melatonin potentiated the contraction induced by NA in hooded and harp seal arteries to a maximum of about 25% above the resting, pre-stretch level. The selective melatonin receptor antagonist luzindole reduced this potentiation by 80%. Species-specific concentrations of melatonin did not potentiate the NA-effect in the domestic pig or the A-induced contraction in any of the species. The results indicate that melatonin specifically increases the NA-sensitivity of smooth muscles in renal arteries from newborn seals. It is presumed that similar effects may operate in foetal and maternal seals and may influence their circulation during maternal diving.