Norepinephrine Pathways Research Articles

Importance of spinal noradrenergic pathways in cardiovascular reflexes and central actions of clonidine and α-methyldopa in the rabbit

We have examined in conscious rabbits the chronic effects of 6-hydroxydopamine (6-OHDA)-induced local lesions of the spinal noradrenaline (NA) pathways on (i) resting mean arterial pressure (MAP) and heart rate (HR), (ii) the nasopharyngeal pressor response, (iii) the sympathetic component of the baroreceptor-heart rate reflex, (iv) the acute responses to intracisternal (i.c.) clonidine and α-methyldopa (α-MD), and (v) the acute NA release response produced by i.c. 6-OHDA. One month after injection of 6-OHDA (40 nmol in 4 μl) into the first cervical spinal cord segment (C 1), the NA content was reduced to 29% in C 2, 45% in T 4 and 61% in L 3 with little non-specific damage. Basal MAP was 14% higher (P < 0.05) than in sham-operated rabbits suggesting increased vasoconstrictor tone. Basal cardiac sympathetic tone was enhanced, but a corresponding increase in cardiac vagal tone resulted in little net effect on resting HR in the spinal NA-depleted group. Spinal NA lesions attenuated the nasopharyngeal pressor reflex by 27% in baroreceptor-intact rabbits and by 38% in sino-aortically denervated (SAD) animals. The lesion did not affect HR range, gain and BP 50 of the sympathetic baroreflex. In SAD rabbits, the acute MAP responses to i.c. 6-OHDA (early hypotension, late hypertension) were not affected by spinal NA depletion, but the early fall in HR (cardiac sympathetic inhibition) was abolished. The hypotension produced by i.c. clonidine or α-MD was not affected by the lesion, probably because many of the NA terminals in the lower thoracic and upper lumbar cord were still intact. Our results suggest that intraspinal NA fibers have a tonic inhibitory action on spinal preganglionic vasoconstrictor and cardiac motoneurons. The spinal NA neurons affecting vasomotor tone (but not cardiac sympathetic tone) are in turn inhibited by higher vasomotor centers receiving projections from the arterial and trigeminal afferents and thereby participate in vasoconstrictor reflexes.

Ibotenic acid-induced nigral lesion and limbic seizure in cats

The degenerative lesions of bilateral substantia nigra (SN) were made by means of microinjections of ibotenic acid (IBO) solutions and influences upon kainic acid (KA)-induced seizures were studied. IBO solutions were injected bilaterally into SN and circumscribed degenerative lesions were made within the bilateral SN. In a control group, phosphate buffer solutions were injected in the bilateral SN. More than one week after the procedures, 1 μg KA was injected into unilateral amygdala via implanted cannula and limbic seizures were induced. The descruction of SN accelerated the secondary generalization of the limbic seizure within 3 days after the KA injection while no secondary generalization was observed within 3 days in the control group. The fact suggests that GABAergic systems of the substantia nigra and/or ascending norepinephrine pathway have a strong inhibitory influence on the secondary generalization of limbic seizures. The hypothesis of nigral mechanisms as a control system for secondary generalization is discussed.

The effects of intracerebroventricularly injected corticotropin-releasing factor (CRF) on the central nervous system: Behavioural and biochemical studies

Intracerebroventricularly (ICV) administered corticotropin-releasing factor (CRF) produces behavioural activation in rats. The present study was designed to investigate the effects of ICV administered CRF on not only locomotor activity, but also the turnover rates of dopamine (DA) and norepinephrine (NE) in various discrete brain regions in rats. ICV administration of 1 μg CRF produced a significant increase in locomotor activity, while ICV administration of 10 μg CRF caused slow stereotypy with prominent grooming. The 3,4-dihydroxyphenylacetic acid/DA ratio, e.g. DA utilization, was increased in the frontal cortex (FC), striatum, hippocampus (HIPP) and amygdala. DA utilization in the FC increased in a dose dependent manner, suggesting that the hyperactivity of the mesocortical DA system is relevant to the grooming response. The 3-methoxy-4-hydroxy-phenylglycol/NE ratio was increased in the FC and HIPP, indicating the involvement of the dorsal NE pathway in ICV CRF-induced behavioural changes in rats. These findings are discussed in connection with the hypothesis that CRF produces behavioural changes consistent with increasing emotionality, especially anxiety, and may serve as a neuroendocrine modulator of stress-enhanced behaviour.

Involvement of spinal monoaminergic pathways in antinociception produced by substance P and neurotensin in rodents

The antinociceptive effects of substance P and of neurotensin have been determined in rodents after depletion of serotonin (5-HT) or noradrenaline (NA) in the spinal cord. The antinociceptive effect of substance P, given intraventricularly, in rats and mice was blocked after depletion of 5-HT in the spinal cord with the neurotoxin 5,7-dihydroxytryptamine (5,7-DHT) or with the inhibitor of the synthesis of 5-HT, p-chlorophenylalanine (PCPA), but not after depletion of NA in the spinal cord with the neurotoxin 6-hydroxydopamine (6-OHDA). Conversely, the antinociceptive effect of neurotensin in mice was blocked after lesion of spinal NA pathways with 6-OHDA. When 5-HT spinal pathways of mice were lesioned with 5,7-DHT, neurotensin-induced antinociception was blocked 7 but not 15 days after the lesion. p-Chlorophenylalanine failed to prevent this effect of neurotensin. The results suggest that the antinociceptive effect of substance P depends on the integrity of spinal 5-HT neurones, whereas that of neurotensin depends on spinal NA neurones and, only to a limited extent, on 5-HT neurones. It seems that different descending systems are involved in the antinociception elicited by these two neuropeptides.

Lesions of the substantia nigra retard Pavlovian eye-blink but not heart rate conditioning in the rabbit.

Bilateral radio-frequency (RF) lesions of the substantia nigra retarded Pavlovian eye-blink (EB) conditioning without affecting concurrent heart rate (HR) conditioning. Dopamine (DA) depletion occurred only in the caudate nucleus, whereas norepinephrine (NE) depletion was limited to the hypothalamus. Bilateral 6-hydroxydopamine (6-OHDA) lesions of substantia nigra retarded acquisition of both EB and HR responses. Six-OHDA lesions produced significant NE depletion in the nucleus acumbens/septi, frontal cortices, and hypothalamus, as well as DA depletion in the caudate nucleus. Trials required to reach EB conditioning criterion was significantly correlated with caudate DA levels. The magnitude of conditioned bradycardia was on the other hand significantly correlated with hypothalamic NE levels. These results suggest that interruption of the nigro-striatal dopaminergic pathway retards Pavlovian somatomotor learning without affecting concurrent autonomic learning, although the latter may depend on an intact ascending NE pathway to the hypothalamus, which passes through the tegmentum and thus is also destroyed in some cases by substantia nigra lesions.

Enhancement of central transmission to sympathetic preganglionic neurons by phosphodiesterase inhibitors and its prevention by clonidine

The effects of 3 phosphodiesterase inhibitors, aminophylline, isobutylmethylxanthine (IBMX), and RO 20-1724, were tested on descending intraspinal and spinal reflex transmission to sympathetic preganglionic neurons in unanesthetized spinal cats. Sympathetic discharges, recorded from upper thoracic preganglionic white rami, were evoked by stimulation (0.1 Hz) of descending excitatory pathways in the cervical dorsolateral funiculus (intraspinal) or of adjacent intercostal nerves (spinal reflex). Each phosphodiesterase rapidly and markedly enhanced transmission through intraspinal pathways but only slowly and modestly enhanced transmission through spinal reflex pathways. Pretreatment with a methyltyrosine-reserpine combination, chlorpromazine, or prazosin markedly restricted the enhancement of intraspinal transmission by IBMX to levels typically produced on spinal reflex pathways. Clonidine markedly depressed transmission through both pathways and prevented enhancement by the phosphodiesterase inhibitors. Yohimbine or tolazoline antagonized the depressant effects of clonidine and restored the ability of the phosphodiesterase inhibitors to enhance transmission. Somatic spinal reflexes were not affected by the phosphodiesterase inhibitors. The results suggest that descending norepinephrine pathways to sympathetic preganglionic neurons activate adenylate cyclase to generate cyclic AMP which increases neuronal excitability, possibly by phosphorylating membrane proteins. Clonidine appears to depress neuronal excitability by inhibiting adenylate cyclase through activation of α 2-adrenergic receptors.

Role of ascending and descending noradrenergic pathways in the antinociceptive effect of baclofen and clonidine

Baclofen and clonidine interact with central noradrenaline (NA) pathways by a variety of mechanisms. The specific role of ascending and descending pathways in antinociception produced by these agents was examined by lesioning the dorsal bundle (DB), locus coeruleus (LC) and descending NA pathways by the microinjection of the neurotoxin 6-hydroxydopamine (6-OHDA). Lesions were verified using high-performance liquid chromatography analysis of NA. Both baclofen and clonidine were injected intraperitoneally in all experiments. The antinociceptive effect of baclofen in the tail-flick test was inhibited 7–21 days after DB lesions. This manipulation decreased NA levels in cortex, hippocampus and hypothalamus but did not alter spinal cord levels. Lesions of the LC potentiated the effect of baclofen 12–16 days postlesion. NA levels were reduced in all the regions just mentioned. DB lesions produced a transient decrease in the effect of clonidine, being observed 7 but not 12–16 days postlesion. Neither acute depletion of NA levels with α-methyl-p-tyrosine (α-MPT), nor LC lesions significantly affected antinociception produced by clonidine. Intraspinal 6-OHDA potentiated the antinociceptive action of clonidine in the tail-flick test. This treatment markedly reduced spinal cord NA levels, but had minimal effects on brain NA. The results of this and previous studies in this laboratory suggest that the antinociceptive effect of baclofen is mediated by interactions with both ascending and descending NA pathways. These pathways appear to interact in a complex manner. Interpretation of data for clonidine is complicated because lesions can both deplete endogenous NA as well as inducing postsynaptic supersensitivity of α 2-receptors . Clonidine does not depend on endogenous NA pathways for producing antinociception because acute depletion of NA with α-MPT does not alter its action. Spinal sites of action are of importance following systemic clonidine because intraspinal 6-OHDA produces supersensitivity. Altering NA activity in ascending pathways alone produces a transient inhibition of the effect of clonidine, but supersensitivity is not apparent. Simultaneous lesions of both ascending and descending pathways do not produce supersensitivity, again suggesting important interactions between such pathways can occur.

Relative activity of metabolic pathways for norepinephrine in endogenous depression

Thirteen patients with endogenous depression, compared to 25 normal controls, had a significantly greater ratio of the urinary excretion of norepinephrine plus its metabolite normetanephrine to either the sum of the two urinary norepinephrine metabolites 3-methoxy-4-hydroxyphenylglycol plus vanillylmandelic acid or to the sum of urinary norepinephrine and all of its metabolites. As urinary levels of norepinephrine and normetanephrine are derived from an extraneuronal metabolic pathway, while levels of 3-methoxy-4-hydroxyphenylglycol and vanillylmandelic acid are more representative of total norepinephrine metabolism, these results suggest that there is a shift in endogenous depression to extraneuronal metabolic pathways for norepinephrine and its metabolites.

Path analysis of psychopharmacological data: Catecholamine breakdown in man

This article applies path analysis to the problem of characterizing metabolic alterations in catecholamine systems when only static or single measures of precursor and products are available. In this article, simulated data from kinetic models of norepinephrine metabolism were examined using path analysis. The method of path analysis, which is based on correlations between compartments and not actual flow, correctly identified operative and nonoperative metabolic pathways in simulated models. In addition, actual data from normal subject as to the 24-hour urinary values for norepinephrine, normetanephrine, vanillymandelic acid, and 3-methoxy-4-hydroxyphenylglycol were analyzed using path analysis. It was found that metabolic routes that are known to be incomplete did not fit these data, whereas the known metabolic pathways for norepinephrine were correctly identified using path analysis. These results suggest that path analysis may provide a useful tool in identifying normal and abnormal catecholamine metabolic pathways in experimental situations where only single values of precursor and products are available.

The naples high (NHE) and low excitable (NLE) rat strains: A progressive report

Female sexual behavior was observed in ovariectomized female rats treated with varying doses of estrogen and progesterone, following electrolytic midbrain lesions. Severe deficits in receptivity were produced by lesions aimed at the dorsal and ventral norepinephrine pathways (DV lesions) and by lesions intended to destroy meso-limbic (A-10) dopamine-producing cells. Significant but less profound impairments were seen following lesions aimed at the serotonin-producing raphe nucleus or the nigro-striatal dopamine cells. Systemically administered d-amphetamine was found to increase receptivity in severely impaired A-10 and DV animals. The observed deficits may result from damage to midbrain structures involved in the sensory-motor or hormonal mediation of the lordosis reflex, or from disruption of a neurochemical system involved in control of lordosis.

Heart receptors for VIP, PHI and secretin are able to activate adenylate cyclase and to mediate inotropic and chronotropic effects. Species variations and physiopathology

We have assessed the presence of VIP/PHI/secretin receptors in heart by: (1) testing the ability of the corresponding peptides to activate adenylate cyclase in cardiac membranes from rat, dog, Cynomolgus monkey and man, and (2) examining the ability of the same peptides to exert inotropic and chronotropic effects on heart preparations from rat and Cynomolgus monkey in vitro. Based on their affinity for natural peptides and synthetic analogs, two types of VIP/PHI/secretin receptors were characterized: the relatively nonspecific “secretin/VIP receptor” of rat heart (that is “secretin-preferring” only in that secretin was more efficient than VIP in stimulating adenylate cyclase), and the “VIP/PHI-preferring” receptor of man, monkey and dog heart. Four physiopathological situations affecting secretin/VIP receptors in rat heart were explored: In male rats from the Okamoto strain and the Lyon strain, two strains presenting spontaneous hypertension, heart membranes exhibited a markedly decreased response of adenylate cyclase to secretin/VIP, with lesser alterations in the responses to isoproterenol and glucagon. This impairment developed in parallel with the occurrence of hypertension and was reproduced in normotensive rats submitted to chronic isoproterenol treatment (but not in Goldblatt hypertensive rats). These findings are consistent with a hyperactivity of norepinephrine pathways in spontaneously hypertensive rats, leading to a reduced number of cardiac post-junctional secretin/VIP receptors bound to adenylate cyclase. Heart membranes from genetically obese (fa/fa) Zucker rats also exhibited severely decreased responses to secretin/VIP with lesser alterations in the responses to glucagon and isoproterenol. These anomalies were specific for the heart, and developed in concomitance with obesity. The first anomaly could not be corrected by severe food restriction. Secretin stimulation of heart adenylate cyclase was also selectively altered in streptozotocin-diabetic rats. Thus, two types of diabetic cardiomyopathy were characterized by a severe local alteration of secretin/VIP receptors coupled to adenylate cyclase. Hypothyroidism, provoked in rat by thyroidectomy or propylthiouracil treatment, again induced a marked decrease in secretin-stimulated cardiac adenylate cyclase activity. In rat papillary muscle electrically stimulated in vitro, secretin exerted a positive inotropic effect. This effect was reduced in obese (fa/fa) Zucker rats. In rat right atrium, secretin also exerted a positive chronotropic effects. Based on a comparison of dose-effect curves of secretin- and isoproterenol-activation of adenylate cyclase on the one hand, rate and contractility on the other hand, we conclude that secretin might directly modulate the mechanical properties of rat heart by increasing cyclic AMP. At variance with rat heart membranes, atrial and ventricular membranes from dog, Cynomolgus monkey, and man were endowed with “VIP-preferring” receptors coupled to adenylate cyclase, on which secretin was virtually devoid of biological activity. In Cynomolgus monkey, the in vitro inotropic and chronotropic effects of VIP were compared to those of isoproterenol on the spontaneously beating right atrium, the electrically stimulated left atrium, and ventricular papillary muscle. The data show direct mechanical effects of VIP that might well be mediated by adenylate cyclase activation as was the case for secretin in rat heart.

Spinal 5-HT or NA uptake inhibition potentiates supraspinal morphine antinociception in rats.

Spinal injection of the specific uptake inhibitor of 5-hydroxytryptamine (5-HT), citalopram, or of noradrenaline (NA), desipramine, potentiated the antinociception following intracerebroventricular injection of morphine in rats tested on the hot plate. Combined spinal injection of citalopram and desipramine caused a synergistic potentiation. The unselective and less potent inhibitor of both 5-HT and NA uptake, amitriptyline, did not cause potentiation. The results support the suggested involvement of both 5-HT and NA pathways in supraspinal morphine antinociception.

Brain noradrenaline and anaesthesia: Behavioural and electrophysiological evidence

Neonatal administration of 6-hydroxydopamine to rat pups was used to deplete brain noradrenaline in the locus coeruleus projection system to less than 5% of normal and the response to barbiturate and non-barbiturate anaesthetics examined. The sleeping time in response to administration of thiopentone, pentobarbitone, methohexitone or hexobarbitone was markedly increased in 6-hydroxydopamine-treated rats, as it was for the non-barbiturates chloral hydrate and diisopropylphenol. The sleeping time for other non-barbiturates such as althesin, ketamine and ethyl carbamate (urethane) was not affected in noradrenaline-depleted rats. Similarly, an index of the evoked potential, recorded in the primary somatosensory cortex to supramaximal electrical stimulation of the forepaw, decreased more markedly with increasing doses of thiopentone in 6-hydroxydopamine-treated rats than in controls. Potentiation of the effect of diisopropylphenol on the evoked cortical response was also seen in noradrenaline-depleted rats while the effect of althesin did not differ. It is suggested that brain noradrenaline pathways originating from the locus coeruleus may play an important role in the duration and depth of anaesthesia resulting from barbiturate and some related agents.

Studies on the mechanisms by which clenbuterol, a β-adrenoceptor agonist, enhances 5-HT-mediated behaviour and increases metabolism of 5-HT in the brain of the rat

The head twitch response in mice produced by injection of 5-hydroxytryptophan (100 mg/kg i.p.) and carbidopa (25 mg/kg i.p.) was enhanced by administration of clenbuterol (0.5 mg/kg i.p.), a β-adrenoceptor agonist. Clenbuterol also enhanced the hyperactivity syndrome in rats produced by quipazine (25 mg/kg i.p.), a 5-hydroxytryptamine (5-HT) agonist. This enhancement was not prevented by depletion of 5-HT in brain with p-chlorophenylalanine or after pretreatment with prazosin. The behavioural responses of the rats to administration of the α 2-adrenoceptor agonist, clonidine, was unaltered by acute or longer-term administration of clenbuterol. Following chronic administration of clenbuterol (5 mg/kg daily for 14 days), a procedure resulting in down-regulation of central β-adrenoceptors, a larger dose of clenbuterol was necessary to enhance the quipazine-induced hyperactivity, suggesting that the mechanism of enhancement involved central post-synaptic β-adrenoceptors. Further evidence for this conclusion was that a lesion of central noradrenaline pathways produced by 6-hydroxydopamine did not abolish the clenbuterol-induced enhancement of the quipazine-mediated behaviour. The binding characteristics of 5-HT 2-receptors were unchanged by acute or chronic administration of clenbuterol. Clenbuterol (5 mg/kg) increased the percentage of plasma free (non-albumin bound) tryptophan, plasma free fatty acid concentration and the concentration of tryptophan and 5-hydroxyindoleacetic acid (5-HIAA) in the brain. The increase in 5-HT turnover in brain was prevented by pretreatment with the β 1-adrenoceptor antagonist atenolol, which enters the brain poorly. It is therefore suggested that the clenbuterol-induced increase in 5-HT metabolism results from the increase in the concentration of plasma free fatty acid which increases plasma free tryptophan and thus increases the concentration of tryptophan in brain and 5-HT synthesis in brain. The clenbuterol-induced enhancement of 5-HT-mediated behaviour is therefore not associated with its effect on 5-HT metabolism. The data are discussed in relation to that obtained after administration of antidepressant drugs.

Functional roles of monoaminergic pathways to sympathetic preganglionic neurons.

Despite considerable progress in mapping the central monoaminergic pathways to sympathetic preganglionic neurons in the spinal cord, the respective functional roles of these pathways have not been resolved. Evidence for both excitation and inhibition has been advanced for each of the three monoamines, serotonin, norepinephrine, and epinephrine. Our previous studies on spinal sympathetic pathways to sympathetic preganglionic neurons support the prevailing opinion that serotonin pathways are inhibitory but did not satisfactorily resolve the functional role of the norepinephrine pathways. However, more recent studies showing that intraspinally evoked sympathetic discharges were rapidly and markedly enhanced by phosphodiesterase inhibitors and that this effect was prevented by clonidine have led to formulation of a coherent hypothesis which accommodates much of the conflicting evidence regarding norepinephrine. In addition, evidence for the role of the epinephrine pathways has been obtained by using a selective inhibitor of epinephrine synthesis. The results of these recent studies complement our previous results and suggest that the excitability of sympathetic preganglionic neurons is regulated by excitatory norepinephrine pathways and inhibitory epinephrine pathways that activate or suppress adenylate cyclase to control intraneuronal levels of cyclic AMP.

Alcohol intake and ethanol intoxication in the rat: Effect of a 6-OHDA-induced lesion of the ascending noradrenaline pathways

The ascending noradrenaline (NA) pathways were lesioned by injecting 6-hydroxydopamine (6-OHDA) 16 μg/ 4 μl bilaterally into the posterior mesencephalon in male Long Evans rats. Another group of rats was pretreated with protriptyline (25 mg/kg), a NA uptake blocking agent, 15 min before they received the intracerebral injections of 6-OHDA. The controls received the vehicle only. Spectrofluorimetric determination of the catecholamine concentrations in various parts of the brain revealed a marked degeneration of the ascending NA systems in the group receiving 6-OHDA. Unexpectedly, the DA systems were also affected by the 6-OHDA treatments. Three weeks after the operation the 6-OHDA group showed a transient increase in ethanol intake. In the tilting-plane test, ethanol (2 g/kg, i.p.) impaired the performance of the 6-OHDA-treated rats significantly more than that of the controls. In contrast, the hypothermic effect of ethanol (4 g/kg, i.p.) was significantly smaller in the lesioned rats. Furthermore, the catecholamine levels in various parts of the brain could be significantly correlated with both the extent of ethanol intoxication and the hypothermia. However, the duration of ethanol-induced narcosis (4 g/kg, i.p.) was not affected by the present treatments. These results give further support for the view that the central NA neurons are important in the control of ethanol intake, and that they are also involved in the expression of the acute effects of ethanol administration.

Effects of brain monoamine depletions on thermoregulation in rabbits.

Biochemical analyses revealed that 5,7-dihydroxytryptamine- (5,7-DHT) treated and 6-hydroxydopamine- (6-OHDA) treated rabbits, respectively, had a significant reduction in diencephalic 5-hydroxytryptamine (5-HT) and norepinephrine (NE) with minimal alterations in the other monoamines. Both 5,7-DHT-treated and 6-OHDA-treated animals, although showing the maintenance of normal body temperatures, did exhibit specific alterations in the thermoeffector outputs. The 5-HT-depleted animals displayed an increase in ear skin blood flow, respiratory heat loss, and metabolism at both ambient temperatures (Ta) of 22 and 2 degrees C. The NE-depleted animals displayed a decrease in metabolism at all levels of Ta tested. Also, the peripheral temperature threshold for the onset of heat loss responses were displaced to higher values than in control animals. In addition, the prostaglandin E1-induced fever was attenuated after the pretreatment of rabbits with either 5,7-DHT or 6-OHDA. The data indicate that brain levels of 5-HT and NE alter, in an apparently reciprocal fashion, the thermoregulatory responses of rabbits. Also, the integrity of 5-HT and NE pathways in brain is vital for the full functioning of a prostaglandin in producing a fever.

Locus coeruleus lesions: Learning and extinction

Bilateral destruction of the nucleus locus coeruleus (LC) by the electrolytic lesion produced large depletions of noradrenaline in the cerebral cortex (85%), hippocampus (70%), and cerebellar cortex (83%), and produced a disruption of the acquisition of an L-shaped runway task for food reward, however, acquisition of another learning task, a spatial discrimination in the same apparatus was not impaired. Acquisition of two other operant learning tasks, a food rewarded, lever press response and a successive visual discrimination, were similarly unaffected. No alteration in extinction behavior was found in the runway or in the food rewarded, lever press response. LC rats did habituate more slowly to distracting stimuli in the runway, but no difference could be found between the lesion and sham animals in locomotor activity in photocell cages or in an open field. These results are compared with the behavioral effects observed after 6-hydroxydopamine lesion to the ascending noradrenaline pathways of the central tegmental tract, and suggest that LC lesions produce a markedly different profile of behavioral alterations.

The effects of 5,7-dihydroxytryptamine-induced lesions of the ascending 5-hydroxytryptamine pathways on the sleep-wakefulness cycle

Summary The role of ascending 5-hydroxytryptamine (5-HT) systems in sleep and waking in the rat was investigated by making fairly selective lesions of these systems with the neurotoxin 5,7-dihydroxytryptamine (5,7-HT). 5,7-HT (4 μg/4 μl) was injected bilaterally into the dorsomedial mesencephalic tegmentum close to the 5-HT pathways. Another group of rats was pretreated with chlorimipramine (25 mg/kg, i.p.) 10 min before they received the intracerebral injections of 5,7-HT. The controls received the vehicle only. The EEG and EMG were continuously recorded for 2–4 postoperative days in order to measure the time the animals spent awake and in different sleep stages. The EEG recordings exhibited both in the 5,7-HT group and in the chlorimipramine-5,7-HT group an increase in waking activity and a corresponding decrease in slow wave sleep 2 (SWS 2) characterized by continuous high voltage slow waves. The changes were significant during 3–4 postoperative days. Unspecific neuronal damage (nerve cell degeneration, glial reaction) was observed at the site of 5,7-HT and vehicle injection (0.6 mm × 1.0 mm and 0.4 mm × 0.6 mm, respectively) but not in the region of the raphe, where the main 5-HT bundle ascends. Accumulations of 5-HT and noradrenaline (NA) were observed in the ascending 5-HT and NA pathways, respectively. The uptake-retention of [ 3 H]5-HT and [ 3 H]NA in the cortical slices and hypothalamic homogenates, predicting the degree of degeneration of 5-HT or NA nerve terminals, was correlated to the different EEG states. When [ 3 H]5-HT uptake in the cortical slices was considered, a significant negative intraindividual correlation was obtained between [ 3 H]5-HT uptake and SWS 1 time. Furthermore, the amount of cortical 5-HT terminals was positively correlated with the time spent in SWS 2 and paradoxial sleep (PS). The studies on the uptake of [ 3 H]NA showed that the NA neurons were also affected by the 5,7-HT treatment. Unexpectedly, chlorimipramine was not found to protect the 5-HT nerve terminals from degeneration, whereas the NA terminals were protected. The present results give further support for the view that the ascending 5-HT neurons are important for the maintenance of SWS 2.

Involvement of central catecholamines in the feedback actions of 17β-estradiolbenzoate on luteinizing hormone secretion in the ovariectomized female rat

(1) Evidence has been presented, based on quantitative microfluorimetric estimations of dopamine (DA) and noradrenaline (NA) levels and turnover, and on radioimmunological measurements of serum luteinizing hormone (LH), and follicle-stimulating hormone (FSH) and prolactin levels, that the central inhibitory feedback action of estradiol on LH secretion mainly involves a marked increase in DA turnover of the lateral palisade zone (LPZ) of the median eminence and also involves a reduction of NA turnover, mainly located in the medial preoptic area (MPOA) and in the subependymal layer (SEL). (2) The mechanism for these changes in catecholamine (CA) turnover is discussed. The possibility is favored that they involve the stimulation of central cytosol estrogen receptors which may even be located in the arcuate DA and possibly reticular NA cell bodies themselves. (3) The marked and long-lasting hypersecretion of prolactin caused by estrogen could be involved in mediating the sustained increase of DA turnover in the median eminence and the sustained reduction of NA turnover. (4) The central facilitatory feedback action of estrogen, on the other hand, may be mainly responsible for the sharp increase of NA turnover in the MPOA and SEL and also the associated reduction of DA turnover in the LPZ in the critical period of both adult cyclic rats and of immature female rats treated with PMS. (5) We take the view that these latter NA and DA turnover changes take precedence over cytosol estrogen receptors, which previous evidence indicates (see Sawyer, 1975) are located in preoptic and amygdaloid regions. (These estradiol concentrating neurons then directly and/or indirectly make connections with the NA and DA pathways). (6) FSH secretion is not controlled by DA and NA pathways. (7) The hypothesis given above is based on the assumption that different estradiol concentrating neurons are involved in the central inhibitory and facilitatory feedback action of estradiol and that the estrogen receptors of these respective neurons have differences which allow their differential activation, the inhibitory feedback leading mainly to a marked increase in the ratio DA activity/NA activity in the median eminence, whereas the facilitatory feedback causes a marked reduction of this ratio.