Effects of catecholamines and β-alanine on tanning of pupal cuticle of Manduca sexta in vitro

When epidermis from wandering stage tobacco hornworm (Manduca sexta) larvae was exposed to 5 μg/ml 20-hydroxyecdysone for 3 days, then exposed to hormone-free Grace's medium, the newly formed pupal cuticle tanned slowly up to 35% of its area by day 12. The addition of 1.3 mM dopamine on either day 4 or 5 slightly increased the area tanned and addition of β-alanine (to 11.2 mM) on days 3–5 enhanced tanning 2–2.5-fold by day 8. Later addition had no effect. When pharate pupal cuticle about 24 h before ecdysis was explanted to Grace's medium, little tanning occurred in 24 h unless dopa or dopamine or their derivatives were added; β-alanine up to 4.4 mM had no effect. Partial tanning occurred in 10 mM dopa or dopamine. More effective were N-β-alanylnorepinephrine and N-β-alanyldopamine which produced nearly maximal tanning at 1 and 5 mM respectively. Up to 10 mM N-β-acetylnorepinephrine had little effect. Thus, dopamine and β-alanine are important to cuticular tanning in vitro and apparently need to be incorporated into the exocuticle during its synthesis. Maximal tanning of this exocuticle then requires further incorporation of β-alanyl conjugates.     

3,4-Dihydroxyphenylalanine (dopa) decarboxylase activity in the arthropod nervous system

1. When homogenates of brains from mature adult locusts (Locusta migratoria) were incubated with l-3-(3,4-dihydroxyphenyl)[3-(14)C]alanine the major radioactive metabolite was dopamine, suggesting the presence of a dopa (3,4-dihydroxyphenylalanine) decarboxylase. 2. Decarboxylation of l-dopa by this tissue, measured under optimum conditions by a radiochemical method, was 21mumol of CO(2)/h per g wet wt. Apparent decarboxylation of l-tyrosine proceeded at 0.34mumol of CO(2)/h per g wet wt. There was no detectable decarboxylation of l-tryptophan, l-histidine or l-phenylalanine. 3. Dopa decarboxylase activity was found in all major regions of the ventral nerve cord of the mature locust (range: 4-7mumol of CO(2)/h per g wet wt.) but was low or absent in thoracic peripheral nerve. 4. Marked decarboxylation of l-dopa was found in homogenates of brains of four other species of insects, and in brain and ventral nerve cord, but not in the claw nerve, of the crayfish. 5. The activity of the locust brain enzyme may be slightly lower at the time of imaginal ecdysis than during the mature period. By contrast, the dopa decarboxylase that produces dopamine as an intermediate in cuticle biosynthesis is known to be high in activity at the time of ecdysis and low in activity during the intermoult stages.     

Glutathione transferase M2-2 catalyzes conjugation of dopamine and dopa o-quinones

Human glutathione transferase M2-2 prevents the formation of neurotoxic aminochrome and dopachrome by catalyzing the conjugation of dopamine and dopa o-quinone with glutathione. NMR analysis of dopamine and dopa o-quinone-glutathione conjugates revealed that the addition of glutathione was at C-5 to form 5-S-glutathionyl-dopamine and 5-S-glutathionyl-dopa, respectively. Both conjugates were found to be resistant to oxidation by biological oxidizing agents such as O(2), H(2)O(2), and O(*-)(2), and the glutathione transferase-catalyzed reaction can therefore serve a neuroprotective antioxidant function.     

Inactivation of creatine kinase induced by dopa and dopamine in the presence of ferrylmyoglobin

We investigated the effect of dopa and dopamine on creatine kinase (CK) activity in the presence of ferrylmyoglobin (ferrylMb). CK was sharply inhibited by dopa and dopamine in the presence of ferrylMb. Dopa and dopamine markedly promoted the reduction of ferrylMb to metmyoglobin (metMb). The semiquinone from dopa and dopamine may be involved in CK inactivation. During inactivation of the enzyme, both kinetic parameters Vmax and Km changed. In addition, reduced glutathione restored the activity of CK at an early stage. These results suggest that inactivation of CK is dominantly due to oxidation of sulfhydryl (SH) groups of the enzyme. Other catechols, such as adrenaline and noradrenaline, little inactivated CK activity, whereas they promoted the reduction of ferrylMb to metMb. Other SH enzymes, including alcohol dehydrogenase (ADH) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), were inactivated to a lesser extent by dopa and dopamine in the presence of ferrylMb. Adrenaline and noradrenaline did not significantly prevent the inactivation of ADH and very slightly inhibited GAPDH. These results suggest that dopa and dopamine act as prooxidants to inactivate SH enzymes in the presence of ferrylMb.     

SEQUENTIAL ANALYSIS: MANGANESE, CATECHOLAMINES AND l-DOPA INDUCED DYSKINESIA

Abstract— This paper specifies methodology for the sequential determination of manganese and catecholamines in selfsame brain samples and shows correlations between them. Small samples were obtained from five regions of brain of cats that had received either saline or levodopa. The doses of levodopa were varied so that although all animals reacted, some developed dyskinesia while others did not. Each sample was first analyzed nondestructively for manganese and then destructively for dopa and dopamine; thus errors inherent in analysing separate samples, due to the structural heterogeneity of the brain, were avoided. Statistically significant correlations were found (1) between levodopa-induced dyskinesia and the concentrations of dopamine and manganese in some of the regions analysed, and (2) between the concentrations of dopamine and of manganese in the caudates of the cats receiving the highest doses of levodopa.     

Tyrosine and catecholamine metabolism in wild-type Drosophila melanogaster and a mutant, ebony

Microinjection of radioactive tyrosine, dopa, and dopamine into mature larvae of Drosophila revealed that the sclerotization pathway is similar but not identical to that in Calliphora: (a) tyrosine is converted to tyrosine-o-phosphate and not to dopa, and (b) the substrate N-acetyldopamine does not accumulate.Larvae of the mutant ebony appear to be similar to the wild type with respect to tyrosine, dopa, and dopamine utilization. About the time of eclosion, however, ebony has twice as much dopamine as normal. Some implications of this are discussed with reference to the mutant phenotype.     

Regulation of melanization of tobacco hornworm larval cuticle in vitro

When tobacco hornworm larvae (Manduca sexta) are allatectomized 5-6 hr before head capsule slippage in the molt to the fifth (final) larval instar, the new cuticle melanizes 3 hr before ecdysis. After explantation between 7 and 3 hr before the onset of melanization, the new cuticle was found to melanize in vitro in Grace's medium only if beta-alanine was removed. When explanted at the onset of melanization, the presence of beta-alanine had no effect on melanization. The addition of either dopa or dopamine was found to be necessary for complete melanization of pieces explanted before the onset of melanization with 0.3 mM of either dopa or dopamine being optimal. Both of these compounds were incorporated into the cuticular melanin. In this optimal medium, melanization occurred over about a 9-hr period after a 5- to 6-hr lag period presumably required for adjustment to the medium. Fifty ng/ml 20-hydroxyecdysone was found to inhibit melanization of pieces explanted 7 hr but not 3 hr before melanization. The hormone neither inhibited uptake of dopa into the epidermis nor prevented melanization in the cuticle once the prophenoloxidase in the premelanin granules was activated. Therefore, 20-hydroxyecdysone may inhibit the activation of the phenoloxidase in the pre-melanin granules, or may inhibit the incorporation of dopa into the granules.     

Enzymatic and Non-Enzymatic Oxygenation of Tyrosine

Tyrosinase isolated from cultured human melanoma cells was studied for tyrosine oxygenation activity. l -Tyrosine and d -tyrosine were used as substrates and dopa was measured with HPLC and electrochemical detection as the product of oxygenation. Incubations were performed in the presence or absence of dopamine as co-substrate. Oxygenation of l -tyrosine occurred only in the presence of dopamine as co-substrate. No oxygenation of d -tyrosine was found, and we conclude that human tyrosinase is characterised by exclusive specificity for the l -isomer of tyrosine in its oxygenase function. It has recently been suggested that superoxide anion is a preferential oxygen substrate for human tyrosinase. Incubations were therefore performed with l - and d -tyrosine, human tyrosinase, and xanthine/xanthine oxidase in the system, generating superoxide anion and hydrogen peroxide. Considerable formation of dopa was observed, but the quantity was the same irrespective of whether d -tyrosine or l -tyrosine was used as the substrate. Furthermore, formation of dopa occurred in a xanthine/xanthine oxidase system when bovine serum albumin (BSA) was substituted for tyrosinase. Our results provide no evidence that superoxide anion is an oxygen substrate for human tyrosinase. In the incubate containing xanthine/xanthine oxidase, catalase completely inhibited dopa formation, and superoxide dismutase and mannitol each strongly inhibited dopa formation. The results are compatible with hydroxyl radicals being responsible for the formation of dopa, since such radicals may be secondarily formed in the presence of superoxide anion and hydrogen peroxide.     

Secretion of Dopa in Phaeochromocytoma

In a series of 11 cases with phaeochromocytoma, two patients were found in whom the tumour secreted dopa in addition to dopamine, noradrenaline, and adrenaline. These two patients were normotensive in spite of high plasma and urinary levels of noradrenaline and adrenaline. This raises the possibility that dopa may be able to protect against the hypertensive action of noradrenaline. Such a mechanism would also explain the absence of hypertension in many cases of neuroblastoma.     

Effect of dopa-decarboxylase inhibition on Aedes aegypti eggs: evidence for sclerotization

Newly deposited fertilized and unfertilized Aedes aegypti eggs are soft and white. Within a short time they darken and harden. Injection of a potent dopa decarboxylase inhibitor (dl)-3-(3,4-dihydroxyphenyl)-2-hydrazino-2-methylpropionic acid (α-MDH) into females, subsequent to a blood meal, results in oviposited eggs which are pale in colour. Moreover, such fertilized eggs do not hatch. The severity of both effects seems to be positively correlated and is dependent upon the time of α-MDH injection.Extracts of mature ovaries are capable of converting dopa to dopamine in the absence of a pretreatment with α-MDH. Mature ovaries obtained from females who had been previously injected with α-MDH could not accomplish this conversion. The inhibitor does not seem to have any effect on dopa oxidase activity and subsequent melanization. We conclude that dopamine is synthesized by blood-fed females via decarboxylation of dopa by dopa decarboxylase and propose that the normal darkening and hardening of A. aegypti eggs is a result of sclerotization.     

Chiasma-induction and tyrosine metabolism in locusts

The production of a gregarization pheromone has been postulated in locusts, with effects on melanization of the hopper cuticle and increased chiasma frequency during meiosis in the adult on crowding or gregarization. Lack of chiasma-inducing effect of the pheromone on albino strains is correlated with the absence or deficiency of some of the products of the metabolic pathways of tyrosine. Some of these products, commercially obtainable, are the amino acids phenylalanine and tyrosine leading to both the melanization and sclerotization pathways; dopamine formed from dopa in the lastnamed pathway; three products of dopamine i.e. protocatechuic acid, noradrenaline and adrenaline. The injection of solutions of these metabolites into the haemolymph of solitary hoppers has shown that only dopa to some extent but noradrenaline to a large extent are effective in raising chiasma frequency in solitarised individuals of normal-coloured strains of Locusta, while in two albino strains, which differ genetically, the injection of dopa, dopamine, protocatechuic acid and noradrenaline proved effective; phenylalanine was effective in only one of these albino strains, while adrenaline was effective in neither. The chiasma-inducing effect of noradrenaline, common to the three strains, is accompanied in the normal-coloured strain by a greater retention of dark coloration during solitarization and by some attainment of the crowded type of morphometric ratios which is a third physical criterion of gregarization. The genetic blocks to the physical criteria of gregaria in the albino strains lie at the immediate level of dopa production or previous to this reaction; it may be construed that such a block in the solitaria of normal-coloured strains also lies at this early level, in this case being induced by too low a pheromone concentration.     

PARTICIPATION OF DOPAMINE- AND SEROTONIN-RECEPTORS IN THE DISAGGREGATION OF BRAIN POLYSOMES BY l-DOPA AND l-5-HTP

Abstract— It has previously been shown that the disaggregation of brain polysomes and suppression of brain protein synthesis observed in rats given the amino acids l -dopa or l -5-HTP is mediated by the decarboxylation products dopamine and serotonin. Present studies demonstrate that the poly-some disaggregation is caused by the interactions of the monoamines with specific receptor sites. Thus, dopa-induced disaggregation is blocked if rats are pretreated with haloperidol or pimozide (but not methysergide or cyproheptadine), while 5-HTP-induced disaggregation is blocked by methysergide or cyproheptadine (but not by haloperidol or pimozide).
Pretreatment of rats with MK-486, a drug that inhibits dopa decarboxylase in blood vessels and peripheral tissues but not brain, does not block dopa-induced brain polysome disaggregation; hence this disaggregation depends on the interaction of dopamine with receptors in the brain parenchyma. Brain polysomes are not disaggregated in rats given intraperitoneal apomorphine (or intracisternal dopamine). The disaggregation caused by dopa is not reduced in animals pretreated with sufficient intracisternal 6-hydroxydopamine to cause major damage to catecholaminergic nerve terminals.     

Roles of dopa decarboxylase and phenoloxidase in the melanization of the tobacco hornworm and their control by 20-hydroxyecdysone

Summary WhenManduca sexta larvae are allatectomized 5 h before head capsule slippage (HCS) in the final larval molt, the new larval cuticle contains granules that melanize 3 h before ecdysis when the ecdysteroid titer falls (Curtis et al. 1984). In both the epidermis and hemolymph of these allatectomized larvae dopamine was higher than dopa prior to and at the time of melanization. Dopamine also increased in the new cuticle as melanization began. Dopa decarboxylase (DDC) activity increased in the epidermis, cuticle, and fat body beginning 16 h after HCS, with a two-fold greater increase in the epidermis of allatectomized larvae. Both -MDH and -fluoromethyl-dopa inhibited epidermal DDC activity and inhibited melanization in vitro when dopa was used as a precursor. Addition of dopamine to the medium allowed melanization in the presence of the inhibitors. All these results indicate that dopamine is likely the primary precursor of cuticular melanin. The diphenoloxidase in the premelanin granules was activated in vivo between 19 and 21 h after HCS and was found to prefer dopamine to dopa and not to convert tyrosine to melanin. The activation of the prophenoloxidase was inhibited by 20-hydroxyecdysone (20-HE), both in vivo and in vitro, if hormone was given by 16 h after HCS. Infusion of 1.2 g/ml 20-HE into allatectomized larvae for 24 h from HCS prevented both the increase in DDC activity and the activation of the premelanin granules. Although the larvae ecdysed after a 15 h delay, melanization never occurred.Abbreviations -MDH L-3-(3,4 dihydroxyphenyl)-2-hydrazine-methylpropionic acid - -FM-dopa R-S--fluoromethyl-dopa - DCC dopa decarboxylase - 20-HE 20-hydroxyecdysone - JH juvenile hormone - HCS head capsule slippage     

Dopaminergic innervation of the rainbow trout pituitary and stimulatory effect of dopamine on growth hormone secretion in vitro

To elucidate which factors regulate growth hormone (GH) secretion in rainbow trout, dopaminergic innervation of the rainbow trout pituitary along with the action of dopamine in vitro, were studied. Brains with attached pituitaries were double-labeled for putative dopaminergic neuronal fibers and somatotropes, using fluorescence immunohistochemistry. A direct dopaminergic innervation to the proximal pars distalis (PPD) with dopaminergic fibers terminating adjacent to somatotropes was demonstrated. Growth hormone secretion from whole pituitaries was measured in perifusate using a homologous GH-RIA. Dopamine (DA; 10(-7)-2x10(-6) g ml(-1)) increased basal GH secretion, with the GH secretion normalizing again after the DA exposure was halted. When pituitaries were pre-treated with somatostatin-14 (SRIF-14; 10(-12)-10(-9) g ml(-1)), before being exposed to different doses of DA, there was an inhibition of GH secretion which was not reversed after treatment of SRIF-14 was halted, unless DA was added. It is concluded that dopamine can function as a GH secretagogue in the rainbow trout pituitary gland.     

Inhibitory actions of dopamine on Limulus visceral muscle involve a cyclic AMP-dependent mechanism

1. The catecholamines dopamine, epinephrine and norepinephrine were detected in alumina extracts of Limulus midgut tissue using high performance liquid chromatography with electrochemical detection. Moderate levels of norepinephrine (28.2 +/- 2.1 ng/g) and dopamine (24.0 +/- 5.2 ng/g) were detected in the midgut, while epinephrine levels (7.4 +/- 0.9 ng/g) were less. Catecholamines were present in all regions along the longitudinal axis of the midgut, and norepinephrine and dopamine levels were highest in posterior regions. 2. Catecholamines decreased muscle tonus and inhibited spontaneous contractions of the Limulus midgut. Dopamine typically decreased spontaneous midgut activity at doses of 10(-8) M or greater, and produced inhibitory actions on all regions of the Limulus midgut. In some preparations epinephrine and norepinephrine elicited a secondary rhythmicity. The actions of dopamine opposed the excitatory effects produced by either proctolin or octopamine. 3. Catecholamines significantly elevated levels of cyclic AMP in Limulus midgut muscle rings. Dopamine (10(-5) M) increased cyclic AMP with a time course consistent with its physiological effects. Forskolin and several methyl xanthines increased Limulus midgut cyclic AMP levels and mimicked the inhibitory effects of dopamine on the isolated midgut preparation. Cyclic nucleotide analogues also produced dopamine-like effects on the isolated midgut preparation. Inhibition of cyclic nucleotide phosphodiesterase prior to addition of dopamine enhanced the effect of this amine to decrease baseline muscle tension. 4. The inhibitory effects of 10(-5) M dopamine on the midgut persisted in solutions of zero sodium and in the presence of tetrodotoxin. Zero calcium solutions gradually reduced spontaneous midgut activity and the effects of dopamine. Calcium channel blockers did not prohibit dopamine-induced relaxation.(ABSTRACT TRUNCATED AT 250 WORDS)     

A kinetic analysis of Drosophila melanogaster dopa decarboxylase

The kinetic mechanism of dopa decarboxylase (3,4-dihydroxy-L-phenylalanine carboxy-lyase, EC 4.1.1.28) was investigated in Drosophila melanogaster. Based on initial velocity and product inhibition studies, an ordered reaction is proposed for dopa decarboxylase. This kinetic mechanism is interpreted in the context of measured enzyme activities and the catecholamine pools in Drosophila. The 1(2)amd gene is immediately adjacent to the gene coding for dopa decarboxylase (Ddc) and determines hypersensitivity to alpha-methyldopa in Drosophila. Dopa decarboxylase does not decarboxylate alpha-methyldopa and hence does not generate a toxic product capable of inhibiting 1(2)amd gene function. We propose that the 1(2)amd gene is involved with an unknown catecholamine pathway involving dopa but not dopamine.     

Structure of Catecholamine Secretory Vesicles from PC12 Cells

Catecholamine secretory organelles were partially purified from PC12 cells. Measurement of the sedimentation coefficient (540S in 0.32 M sucrose), density in an isoosmotic gradient (1.139 g/cm), and density in an isoosmotic gradient using D2O as a solvent (1.205 g/cm3) have allowed us to calculate the molecular weight (1.17 X 10(9) daltons), radius (74 nm), and water content (62% vol/vol) of the secretory vesicle. The vesicle appears to contain ATP, but the molar ratio of 3,4-dihydroxyphenylethylamine (dopamine) to ATP in the particles is high (16.5) and the ATP was frequently asymmetrically distributed in the vesicle fraction. The particle behaves like a true secretory particle in that the dopamine content of the particle is increased by pargyline, diminished by depolarization, and abolished by reserpine. Sequential purification of PC12 lysates on controlled pore glass columns and isoosmotic Ficoll gradients produced a 20-30-fold purification, but this enrichment is not sufficient to produce a homogeneous population of vesicles. An 82,000-dalton protein copurifies with secretory granules and appears to be the major secreted protein. At this stage of purification this single protein makes up about 30% of the protein in the vesicle-containing fractions and so the vesicles must be approaching homogeneity.     

The Effects of Hydroxyl Radical Attack on Dopa,Dopamine, 6-Hydroxydopa,and 6-Hydroxydopamine

High pressure liquid chromatography with electrochemical detection (HPLC-ED) was employed in conjugation with a sensitive and specific salicylate hydroxylation assay to evaluate the immediate effects of hydroxyl radical (·OH) attack on four catechol intermediates of eumelanin, dopamine (3,4-dihydroxyphenylethylamine), its precursor dopa (3,4-dihydroxyphenylalanine), and their respective neurotoxic trihydroxyphenyl derivatives, 6-hydroxydopamine (2,4,5-trihydroxyphenylethylamine,6-OHDA) and 6-hydroxydopa(2,4,5-trihydroxyphenylalanine, TOPA). Semiquinone and quinone species were identified as the initial products of the oxidation of these four catechol substrates. The enhanced oxidations of the catechols when exposed to ·OH attack was accompanied by marked decreases in the level of each semiquinone species. Quinone levels were elevated in reactions involving ·OH attack on dopamine and 6-OHDA, but absent in reactions involving radical attack on dopa or TOPA, suggesting that dopaquinone (DOQ) and TOPA p-quinone (TOPA p-Q) are oxidized more rapidly by‘OH than are the quinones of dopamine and 6-OHDA. The formation of 6-OHDA p-quinone (6-OHDA p-Q) in incubations involving DA and ·OH suggest that the ·OH-mediated hydroxylation of DA may be a mechanism for generating this potentially cytotoxic trihydroxyphenyl. The results of this study demonstrate for the first time that semiquinone and quinone intermediates of eumelanin are the initial products derived from the ·OH-mediated oxidations of dopa, DA, TOPA, and 6-OHDA. These observations suggest that if ·OH is generated beyond the capabilities of cytoprotective mechanisms, the radical can rapidly oxidize catechol precursors, augment melanogenesis, and generate additional cytotoxic quinoid intermediates of eumelanin.     

The role of tetrahydrobiopterin and catecholamines in the developmental regulation of tyrosine hydroxylase level in the brain

Tyrosine hydroxylase (TH) is a rate‐limiting enzyme for dopamine synthesis and requires tetrahydrobiopterin (BH4) as an essential cofactor. BH4 deficiency leads to the loss of TH protein in the brain, although the underlying mechanism is poorly understood. To give insight into the role of BH4 in the developmental regulation of TH protein level, in this study, we investigated the effects of acute and subchronic administrations of BH4 or dopa on the TH protein content in BH4‐deficient mice lacking sepiapterin reductase. We found that BH4 administration persistently elevated the BH4 and dopamine levels in the brain and fully restored the loss of TH protein caused by the BH4 deficiency in infants. On the other hand, dopa administration less persistently increased the dopamine content and only partially but significantly restored the TH protein level in infant BH4‐deficient mice. We also found that the effects of BH4 or dopa administration on the TH protein content were attenuated in young adulthood. Our data demonstrate that BH4 and catecholamines are required for the post‐natal augmentation of TH protein in the brain, and suggest that BH4 availability in early post‐natal period is critical for the developmental regulation of TH protein level.     

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.