Noradrenaline, dopamine and serotonin levels and metabolism in the human hypothalamus: observations in Parkinson's disease and normal subjects

In order to determine whether, besides the severe striatal dopamine (DA) loss, other brain neurotransmitter changes may be a constant biochemical feature of idiopathic Parkinson's disease (iPD), we measured the concentration of the three major brain monoamines noradrenaline (NA), DA, and serotonin (5-HT) and their metabolites in five rostro-caudal subdivisions of the hypothalamus of eight control patients and nine patients with morphologically confirmed iPD. In the whole hypothalamus of the iPD patients we found a mild to moderate mean reduction of NA(−52%, P < 0.05),DA(−25%),and5-HT(−26%). At the subregional level, the most consistently affected area was the intermediate subdivision of the hypothalamus proper where all three monoamines were statistically significantly reduced. Evaluation of individual patient values indicated that, in contrast to the constant and severe DA reduction present in putamen of each of the iPD patients (DA loss ranging from 96% to 99%), several of these patients had whole (and subregional) hypothalamic monoamine values well within the range of controls. We conclude that, although possibly involved in autonomic and/or endocrine disturbances in some patients with iPD, none of the observed monoamine changes in the hypothalamus is an obligatory feature of iPD. Our study demonstrates the need for evaluation of individual patient values rather than mean differences in order to permit valid conclusions to be drawn as to whether an observed neurochemical change can be regarded as specific to a given brain disorder.     

Effects of a lesion in the dorsal raphe nuclei performed during the juvenile period of the female rat, on puberty

The present study examined the effect of thermic lesions on the dorsal raphe nuclei (DRN) of the female rat, performed at various ages during the prepubertal period (21, 24, or 27 days), on puberty, and at first ovulation. In comparison with sham-operated animals, the age of vaginal opening and first vaginal oestrus was delayed in rats with a DRN lesion performed at the end of the infantile period (day 21) (45.6 ± 0.94 vs. 89.9 ± 1.03, p < 0.05), whereas differences were not observed in animals with lesions performed during the juvenile period (day 24 or 27). The number of ova shed by ovulating animals was greater in those rats with lesions performed on day 24 or 27 (9.7 ± 0.4 vs. 7.4 ± 0.4; 9.5 ± 0.5 vs. 7.7 ± 0.4, p < 0.05). Ovarian follicular atresia in these animals was significantly lower than in control and sham-operated ones. On the day of first vaginal oestrus and 48 h after the lesion, the serotonin-hypothalamic concentration decreased in all lesioned animals. Present results support the idea of the participation of the serotoninergic system, arising from the DRN, in the neuroendocrine mechanisms regulating the onset of puberty and the first ovulation, with variations depending on animal maturity.     

Role of nitric oxide in the regulation of monoaminergic neurotransmission

Data accumulated in the last decade indicate that nitric oxide (NO) participates in the regulation of neurotransmission in the central nervous system. Due to its physicochemical properties, NO is an ideal mediator of nonsynaptic interactions. The importance of monoaminergic systems in the function of the brain is clearly shown by the number of severe neuropsychiatric diseases (e.g. depression, Parkinson's disease) caused by the impairment of monoaminergic neurotransmission. Because of their neuroanatomical characteristic, monoaminergic systems participate mainly in nonsynaptic interactions. Since NO is a potential nonsynaptic modulator, it may have an important role in the regulation of monoaminergic systems. The aim of the present review is to survey the literature on the effect of NO on dopaminergic, noradrenergic and serotonergic neurotransmission. The potential mechanisms of action are summarized. Since there is no agreement in the literature on the nature of the effect of NO exerted on monoaminergic neurotransmission, and there are contradictory data concerning the mechanisms involved, the possible reasons for this unusual inconsistency are also discussed.     

Deprenyl stimulates the efflux of monoamines from the rat hypothalamus in vitro

The direct effects of L-deprenyl, a monoamine oxidase inhibitor, on the hypothalamus of male Sprague-Dawley rats was investigated by measuring the efflux of norepinephrine (NE), dopamine (DA), serotonin (5-HT), dihydroxyphenylacetic acid (DOPAC), and 5-hydroxyindoleacetic acid (5-HIAA) using a combination of high performance liquid chromatography with electrochemical detection and an in vitro incubation system. After measuring basal efflux by incubating the hypothalami with Krebs-Ringers Henseleit (KRH) alone during the first incubation period, hypothalami were incubated either with the medium, KRH alone (0 mM), or KRH containing 0.1, 1, and 10 mM L-deprenyl. During the third incubation period, hypothalami were again incubated with KRH alone to measure the residual effects if any. During the final incubation period, the hypothalami were stimulated with high K⁺ KRH. Deprenyl produced a dose-dependent increase in the efflux of NE, DA, and 5-HT from the hypothalami. Neurotransmitter efflux returned to pretreatment levels when L-deprenyl was removed from the medium. In contrast to NE, DA, and 5-HT, the efflux of the metabolites DOPAC and 5-HIAA was inhibited in a dose-dependent fashion after incubation with L-deprenyl. Results from this study demonstrate that L-deprenyl is capable of stimulating the efflux of neurotransmitters in vitro by a direct action on the hypothalamus.     

BDNF increases monoaminergic activity in rat brain following intracerebroventricular or intraparenchymal administration

We have previously demonstrated alterations in serotonin metabolism within descending pathways following infusion of brain-derived neurotrophic factor (BDNF) into the midbrain, near the periaqueductal gray and dorsal and median raphe nuclei. The aim of the present study was to extend these studies to include a comprehensive regional examination of monoamine (serotonin, dopamine and norepinephrine) and metabolite levels in discrete areas of the intact, adult rat forebrain following direct intraparenchymal midbrain BDNF infusion. We have compared neurochemical changes following midbrain infusion of BDNF to those obtained following intracerebroventricular (i.c.v.) infusion. Significant increases in levels of 5-HIAA and/or the 5-HIAA/5-HT ratio were found in all areas examined including the hippocampus, cortex, striatum, n. accumbens, substantia nigra and hypothalamus following both midbrain and i.c.v. infusion. Changes in dopaminergic activity were also observed, but displayed more regional specificity, i.e. changes were found primarily within the striatum and cortex. The two infusion sites produced similar patterns of neurochemical effects although the magnitude of the changes did vary in some areas. These results suggest that BDNF increased synthesis and/or turnover of serotonin, and to a lesser extent dopamine, in the mature rat forebrain. Furthermore, these data point to possible functional roles for BDNF in neuropsychiatric and neurodegenerative conditions which involve a dysregulation of these monoamine systems.     

Dopaminergic and serotonergic activity in the hypothalamus during early and late pregnancy

Push-pull cannulae were implanted into the arcuate nucleus of pregnant or ovariectomized (OVX) rats, and the perfusate samples were analyzed for biogenic amines and their metabolites. Injection of parygline, a monoamine oxidase inhibitor, resulted in a decrease in 3,4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxyindoleacetic acid (5-HIAA) levels in the samples, and detectable amounts of dopamine (DA) and serotonin (5-HT), which were usually not measurable prior to injection. Administration of 5-hydroxytryptophan resulted in a sharp increase in 5-HIAA and 5-HT levels in the perfusates, and no change in DOPAC levels. Push-pull perfusion was done between midnight and 06.00 h on day 8 and 16 of pregnancy. In those rats which showed a nocturnal prolactin (PRL) surge on day 8, 5-HIAA levels were very high compared to those that did not, or compared to those on day 16, which had chronic low PRL levels. DOPAC levels were not significantly different in the 3 groups. Perfusion of medial basal hypothalamic (MBH) fragments taken from mother rats on day 8 of pregnancy during the PRL surge spontaneously released more DA or 5-HT than did fragments taken on day 16 at the same time of day. These results suggest that serotonergic activity in the MBH is higher on day 8 of pregnancy, in parallel with the occurrence of PRL surges, than on day 16 when no surges are present. Dopaminergic activity, as measured by DOPAC levels in push-pull samples, does not appear to be different between the two days.     

Effect of aging on psychological stress-induced increases in noradrenaline release in the rat anterior hypothalamus: an in vivo microdialysis study

Basal and psychological stress-induced noradrenaline (NA) release were studied by intracerebral microdialysis in the hypothalamus of rats aged 9 weeks or 12 months. Basal NA output was not significantly different between young (9-week-old; 2.11±0.22 pg/20 min) and aged (12-month-old; 2.29±0.34 pg/20 min) rats. Psychological stress for 20 min significantly increased NA release in both groups (186% and 142% of baseline at 9 weeks old and 12 months old, respectively); however, the increase in aged rats was significantly lower than that in young rats (P<0.01). This finding suggests that the noradrenergic neuronal response to psychological stress is attenuated in aged rats.     

Intracerebroventricular injection of prostaglandin E(1) changes concentrations of biogenic amines in the posterior hypothalamus of the rat

Since the posterior hypothalamus (PH) plays a key role in the control of body temperature, the aim of this study was to evaluate the changes in adrenaline, noradrenaline and dopamine levels in the PH during the hyperthermia induced by prostaglandin E(1) (PGE(1)). The concentration of adrenaline, noradrenaline and dopamine in the PH, the firing rate of the sympathetic nerves innervating interscapular brown adipose tissue (IBAT), IBAT and colonic temperatures (T(IBAT) and T(C)) were monitored in 12 urethane-anaesthetized male Sprague-Dawley rats before and after an intracerebroventricular injection of 500 ng PGE(1) dissolved in 2 microl of 0.9% NaCl saline solution or only saline. The catecholamines were collected using a microdialysis probe and quantified by HPLC. The results showed that PGE(1) caused a significant increment in the concentration of adrenaline from 15. 83+/-2.69 to 34.95+/-3.9 ng ml(-1) and of dopamine from 35.15+/-4.48 to 55.68+/-6.21 ng ml(-1). A significant decrease in the level of noradrenaline from 18.75+/-2.05 to 8.56+/-2.26 ng ml(-1) was registered. The firing rate of sympathetic nerves to IBAT was increased from 100+/-0% to 204.83+/-15.22% by PGE(1). T(IBAT) and T(C) rose respectively from 36.91+/-0.15 degrees C to 38.88+/-0.29 degrees C, and from 36.7+/-0.15 degrees C to 38.13+/-0.36 degrees C after the injection of PGE(1). The changes in adrenaline and noradrenaline occurred during the first 20 min as did the changes in temperature and firing rate, while the change in dopamine was delayed until 21-60 min after the PGE(1) injection. No significant change of analyzed variables was found in the control rats. These findings suggest that these biogenic amines of the PH are involved in the control of the sympathetic and thermogenic changes induced by PGE(1).     

Effect of estradiol and testosterone on catechol-O-methyl transferase activity of rat superior cervical ganglion,pineal gland,anterior hypophysis and hypothalamus

Summary The effect of estradiol and testosterone on the activity of catechol-O-methyl transferase (COMT) of the superior cervical ganglion, pineal gland, anterior hypophysis and hypothalamus were examined in castrated rats. In female rats a single injection of 2g of estradiol caused a significant 25% increase of hypothalamic COMT, whereas after 3 daily injections no differences in enzyme activity were observed between hormone- and vehicle-injected controls. In male rats testosterone propionate (0.5 mg) increased hypothalamic COMT by 53%; this effect was not detectable in animals treated with the androgen for 3 days. COMT activity of the superior cervical ganglion, pineal gland and anterior hypophysis did not exhibit significant changes after any of the hormone schedules used.     

Effects of intracerebroventricular injection of glucagon like peptide-1 and its related peptides on serotonin metabolism and on levels of amino acids in the rat hypothalamus

High concentrations of glucagon-like peptide-1 (7-36) amide (GLP-1) and its specific receptor (GLP-1R) have been found in the rat hypothalamus. In this study the actions of GLP-1 and its related peptides, exendin-4 (GLP-1R agonist), exendin (9-39) (GLP-1R antagonist) and GLP-1 (9-36) amide (the major GLP-1 metabolite) on levels of serotonin (5-HT), 5-hydroxyindolacetic acid (5-HIAA) and amino acids (Glu, Asp, Gln, Gly, Tyr, Trp, GABA) in the hypothalamus were investigated. Intracerebroventricular (ICV) injection of GLP-1 (4 nmol) produced a significant reduction in levels of 5-HT (54%) and all measured amino acids (34 to 56%) compared with saline injected controls, whereas exendin (9-39) (4 nmol) was ineffective. ICV injection of exendin-4 produced a significant reduction in the levels of 5-HT, 5-HIAA, Trp, Glu, and Tyr. ICV injection of GLP-1(9-36) amide showed a statistically significant increase in the level of 5-HT, 5-HIAA and all the amino acids tested in this study. Prior administration of exendin (9-39) or GLP-1 (9-36) amide blocked the effects of GLP-1 on the levels of 5-HT and the amino acids. These data are consistent with exendin-4 being a GLP-1R agonist and exendin (9-39) being a specific GLP-1R antagonist. GLP-1 (9-36) amide, a primary metabolite of GLP-1, appears to act as an endogenous antagonist at the GLP-1R.     

Gender-dependent and genotype-sensitive monoaminergic changes induced by polychlorinated biphenyl 153 in the rat brain

Polychlorinated biphenyls (PCBs) are present as ortho- and non-ortho-substituted PCBs, with most of the ortho-substituted congeners being neurotoxic. The present study examined effects of the ortho-substituted PCB 153 on dopamine, serotonin and amino acid neurotransmitters in the neostriatum of both male and female Wistar Kyoto (WKY) and spontaneously hypertensive rat (SHR) genotypes. PCB 153 exposure at p8, p14 and p20 had no effects on levels of these transmitters when examined at p55, but led to increased levels of both homovanillic acid and 5-hydroxyindoleacetic acid, the degradation products of dopamine and serotonin, respectively, in all groups except the female SHR. Immunoblotting showed that PCB exposure induced gender-specific decreases in dopaminergic synaptic proteins. These included a novel finding of decreased levels of the dopamine D5 receptor in both genders and genotypes, whereas male-specific changes included decreases in the postsynaptic density (PSD)-95 protein in the WKY and SHRs and a decrease in the presynaptic dopamine transporter in both the WKY and, less clearly in the male SHR. A female-specific tendency of increased vesicular monoamine transporter-2 was observed in the SHRs after PCB exposure. No changes were seen in tyrosine hydroxylase, the cytoskeletal neurotubulin or the plasma membrane marker Na⁺/K⁺-ATPase in any strain. Hence, PCB-exposure led to increases in monoamine transmitter turnover in both male and female animals, whereas decreases in both pre- and postsynaptic dopaminergic proteins were predominantly seen in male animals. PCB 153 may therefore induce neostriatal toxicity through both presynaptic and postsynaptic mechanisms in both genotypes and genders, including effects on the aspiny interneurons, which employ the D5 receptor to mediate dopamine effects on interneurons in the basal ganglia.     

Corticosterone-dependent alterations in utilization of catecholamines in discrete areas of rat brain

This study investigated the impact of chronic adrenalectomy (ADX), and subsequent corticosterone (CORT) replacement to ADX rats, on brain levels of norepinephrine (NE) and dopamine (DA) and their extent of depletion after -methyl-p-tyrosine (-MpT) administration. Seven discrete hypothalamic areas, namely, the paraventricular nucleus (PVN), medial preoptic nucleus (POM), dorsomedial nucleus (DMN), ventromedial hypothalamus (VMH), perifornical lateral hypothalamus (PLH), supraoptic nucleus (SON), and arcuate nucleus/median eminence (ARC-ME), were examined. The steady-state content of NE and DA in all areas remained essentially unaltered 7 days after ablation of the adrenal glands, as well as after subsequent CORT replacement therapy in ADX rats. However, ADX, which reduced circulating CORT levels to 0.3 μg% as compared to > 3.0 μg% in sham rats, caused a significant increase in the depletion of NE following -MpT treatment, in 4 out of the 7 brain sites examined (PVN, PLH, DMN and ARC-ME). In these brain sites, the NE turnover rate (K,pg/μg protein/h) and rate constant (K,h⁻¹) increased following ADX. The chronic subcutaneous CORT implant (200 mg), which raised circulating CORT levels of ADX rats to 11 μg%, prevented this enhancement of NE turnover in the PVN, PLH and ARC-ME, but not the DMN. Unlike NE, DA utilization in the 7 discrete hypothalamic areas of -MpT-treated rats remained unaltered after ablation of the adrenal glands, as well as after the CORT replacement therapy in ADX rats. These results indicate that circulating CORT may have a modulatory role in the regulation of NE in specific discrete hypothalamic areas, and thereby have an impact on the control of various physiological responses.     

The differential effect of angiotensin II and angiotensin 1-7 on norepinephrine, epinephrine, and dopamine concentrations in rat hypothalamus: the involvement of angiotensin receptors

Angiotensin 1-7 has been recently claimed the active member of the angiotensins' family. In the present study we compared the effect of angiotensin II and angiotensin 1-7 on the concentration of dopamine, serotonin, epinephrine, and norepinephrine and some of their metabolites in the rat hypothalamus, where the levels of angiotensins are particularly high. Intracerebroventricular injection of angiotensin II, but not angiotensin 1-7, time-dependently elevated the levels of both epinephrine (p < 0.05) and norepinephrine (p < 0.05) in the hypothalamus and both effects could be prevented by intracerebroventricular injection of either AT(1) (candesartan), AT(2) (PD123319) or AT(1-7) (A-779) receptor antagonist. Neither angiotensin II nor angiotensin 1-7 produced any changes in the level of dopamine, dihydroxyphenylacetic acid, homovanilic acid, serotonin, 5-hydroxyindoleacetic acid, or tryptophan at any time point in comparison with the control groups. However, AT(1) but not AT(2) receptor blockade, unmasked the stimulatory effect of angiotensin 1-7 on dopamine concentration in the hypothalamus. Thus, angiotensin II and its active metabolite angiotensin 1-7 regulate selectively, albeit differentially, adrenergic, noradrenergic and dopaminergic systems in the hypothalamus, the effects that involve AT(1), AT(2) and AT(1-7) angiotensin receptors.     

Involvement of monoaminergic system in the antidepressant-like effect of the flavonoid naringenin in mice

Dietary flavonoids possess a multiplicity of neuroprotective actions in various central nervous pathophysiological conditions including depression. In this study, the neuropharmacological mechanism of the dietary flavonoid naringenin was investigated in the mouse behavioral models of depression. For this purpose, we investigated the influence of pretreatment with the inhibitors of serotonin or noradrenaline synthesis, p-chlorophenylalanine methyl ester or α-methyl-p-tyrosine, respectively in the anti-immobility effect of naringenin. Compared to the control group, naringenin significantly decreased the immobility time after acute treatment in the mouse tail suspension test (10, 20 and 50 mg/kg), but not in the forced swimming test, without producing locomotor alteration in the open-field test. In addition, pretreatment of mice with p-chlorophenylalanine methyl ester (100 mg/kg) or α-methyl-p-tyrosine (100 mg/kg) prevented the anti-immobility effect of naringenin (20 mg/kg) in the tail suspension test. Taken together, this data demonstrated that naringenin possessed potent antidepressant-like property via the central serotonergic and noradrenergic systems. Thus, our findings suggest the therapeutic potential of this dietary flavonoid in central nervous system disorders especially depression where monoaminergic systems are involved.     

Glucagon injected in the lateral hypothalamus stimulates sympathetic activity and suppresses monoamine metabolism

Glucagon injected in the lateral hypothalamus stimulates sympathetic activity and suppresses monoamine metabolism. The central hypothesis underlying this study is that there is a reciprocal relationship between food intake and sympathetic activity to IBAT. This hypothesis was tested by using intrahypothalamic microinjections of glucagon, a peptide that has been reported to decrease food intake. Sympathetic nerve activity to interscapular brown adipose tissue (IBAT) was measured as electrophysiological discharges of sympathetic nerves to IBAT. The microinjection of glucagon into the lateral hypothalamus (LH) increased sympathetic nerve activity by + 103.8 ± 35.0% (mean±S.E.M.) from pre-injection basal level by 30 min after injection. There was a gradual return to baseline. Micro-injection of glucagon into the LH depressed food intake. Monoamine metabolism was measured by using a microdialysis probe attached to a guide cannula for microinjection of glucagon into the LH. After microinjection of glucagon, the dialysates were collected over 30 min intervals and assayed for norepinephrine (NE), serotonin (5-HT), dopamine (DA) and their metabolites (3-methoxy-4-hydroxyphenylglyucol (MHPG); 5-hydroxyindole-3-acetic acid (5-HIAA); and 3,4-dihydroxyphenylacetic acid (DOPAC). Glucagon suppressed both NE and MHPG concentrations in the lateral hypothalamus (LH), and the concentration of DOPAC was also decreased. There was no change of 5-HT concentration but 5-HIAA levels were reduced by glucagon treatment. These data show that glucagon injected in the LH stimulates sympathetic activity and suggest that this may have occurred by suppression of norepinephrine, dopamine and serotonin turnover in the LH of freely moving rats. These data support the hypothesis of a reciprocal relationship between food intake and sympathetic activity.     

Regulation of α-melanocyte-stimulating hormone release from superfused slices of rat hypothalamus by serotonin and the interaction of serotonin with the dopaminergic system inhibiting peptide release

Release of α-melanocyte-stimulating hormone (α-MSH) from frontal slices of rat hypothalamus superfused with oxygenated artificial cerebrospinal fluid (ACSF) was quantified by radioimmunoassay. Control depolarisations with 50 mM KCl-containing ACSFm produced significant increases in α-MSH release which were partially blocked by 10⁻⁶ M cinnaserin, a serotonin (5-HT) receptor antagonist. Superfusion of the tissues with varying concentrations of 5-HT *10⁻⁷ M to 10⁻⁴ M) resulted in an inverted U-shaped dose-response curve, maximum α-MSH release being obtained with 10⁻⁶ M 5-HT. Addition of 10⁻⁶ M cinanserin shifted the 5-HT dose-response curve to the right whilst the presence of 10⁻⁸ M flupenthixol, a dopamine receptor antagonist, resulted in a sigmoidal 5-HT dose-response curve. Superfusion with ACSF containing either 10⁻⁷ M fluoxetine, a 5-HT re-uptake inhibitor, or 10⁻⁷ M p-chloroamphetamine, an agent releasing 5-HT, induced significant increases in α-MSH release which were abolished in the presence of 10⁻⁶ M cinanserin. These data demonstrate the presence of an endogenous 5-HT system that exerts a biphasic effect on α-MSH release. A stimulatory effect caused by lower 5-HT concentrations appears to be a direct action whilst an inhibitory effect at higher concentrations is mediated through an inhibitory endogenous dopaminergic system. A significant proportion of K⁺-stimulated peptide release is 5-HT-mediated.     

Effects of maternal lead administration on monoaminergic,GABAergic and glutamatergic systems

The effects of perinatal exposure to lead (300 mg/l) on the development of monoaminergic and aminoacidergic systems were evaluated in the striatum, cerebral cortex (Cx), dorsal hippocampus (d-Hipp) and basal-medial hypothalamus. Maternal exposure to lead produced regional alterations in monoamine content, with increases in dopamine and serotonin or their metabolites. Further, decreased glutamate levels were seen in all brain regions studied, while GABA content decreased only in the Cx. Together, these results show that lead causes alterations to neurotransmitter systems during development. These may be related to lead-induced neurobehavioral impairment.     

Estrogen receptors and aromatase activity in the hypothalamus of the female frog, Rana esculenta. Fluctuations throughout the reproductive cycle

It is well known that certain actions of androgen are mediated through in situ aromatization to estrogen in neural target tissues. This study was undertaken to investigate androgen utilization in the hypothalamus of the female frog, Rana esculenta, through a quantification of estrogen receptors and aromatase activity during the reproductive cycle. ³H-estradiol-binding molecules were present in both the cytosol and the nuclear extract of the hypothalamus. These molecules bound specifically ³H-estradiol with high affinity (Kd 10⁻¹⁰ M) and low capacity (cytosol: 1.2±0.4 fmol/mg protein; nuclear extract: 7.9±0.6 fmol/mg protein). Aromatase activity was detected in the microsomal fraction of the hypothalamus using a sensitive in vitro radiometric assay. Both aromatase activity and nuclear estrogen receptor binding fluctuated in synchrony throughout the reproductive cycle. Western blot analysis of aromatase protein revealed one immunoreactive band with a molecular weight of approximately 56 kDa. In contrast to aromatase enzyme activity, the relative levels of aromatase protein changed little during the reproductive cycle suggesting that post-translational mechanisms may be involved in regulating estrogen synthesis in the frog brain. A possible role for estrogens in the modulation of the reproductive behavior in this species is suggested.