Cyclic and age-related changes in norepinephrine concentrations in the medial preoptic area and arcuate nucleus

High-performance liquid chromatography with electrochemical detection (HPLC-EC) and Palkovits' microdissection technique were used to measure norepinephrine (NE) concentrations in the medial preoptic area (MPA) and arcuate nucleus (AN) during various stages of the estrous cycle. NE was measured seven times at 2-h intervals between 1000 h and 2200 h on the days of proestrus and diestrus in young (4-month-old) rats and four times at 2-h intervals between 1400 h and 2000 h in old (20–22-month-old) persistently diestrous rats. On the day of proestrus in young animals, NE increased progressively from low levels at 1000 h to peak levels at 2000 h, followed by a sharp decline at 2200 h. In contrast, no changes in NE occurred on the day of diestrus. Unlike the young proestrous rats, but similar to the young diestrous rats, no changes in NE concentrations either in the MPA or in the AN occurred in the old persistently diestrous rats. These data demonstrate that NE concentrations in the MPA and AN change during the estrous cycle. We believe the increase in NE on the aftemoon of proestrus is related to the surge in serum luteinizing hormone (LH) that occurs simultaneously in this stage of the estrous cycle. The lack of change in NE concentrations in the young diestrous and persistently diestrous old animals is consistent with the well-established absence of changes in serum LH in these animals.     

Effect of exogenous dopamine on hypothalamic dopamine and norepinephrine concentrations in the neonatal brain in rats

Hypothalamic dopamine (DA) and norepinephrine (NE) concentrations were studied in the neonatal rats after acute (postnatal day 4) or chronic (postnatal days 1-10) DA injections (0.5 mg in 5% dextrose in 0.45% saline). Acute injection of DA twice on postnatal day 4 resulted in an increase of hypothalamic DA and NE concentrations 16 hr later. Chronic treatment with the DA (twice in a day) for 10 days resulted in a reduction of NE concentration in the hypothalamus. The results of these studies suggest that the amount and duration of exposure to exogenous DA during postnatal development may result in divergent effects on hypothalamic catecholamine concentration.     

In vivo changes in responsiveness of the caudate nucleus tol-DOPA infusion as a function of the estrous cycle

In the present experiment we measured changes in the responsiveness of the nigrostriatal dopaminergic system tol-DOPA as a function of the estrous cycle in the freely behaving animal with push-pull perfusion. Intact female rats were implanted with a push-pull cannula directed at the caudate nucleus and perfused repetitively on each day of their estrous cycle. During the perfusion, the response of the caudate nucleus was tested with two successive increasing doses ofl-DOPA (1.0 and 10 μM) infused directly through the push side of the cannula. In response to 1.0 μMl-DOPA, significantly greater dopamine release was obtained for female rats perfused on proestrus, with all 6 females showing maximal responsiveness at this estrous cycle stage. There was an overall 3–4 fold greater increase in dopamine release to the 10 μMl-DOPA infusion, regardless of estrous cycle day. However, with the use of this 10 μMl_DOPA dose, the significant effect at proestrus was maintained. These results demonstrate that the responsiveness of the caudate nucleus tol-DOPA is maximal at proestrus. These data together with previous work from our laboratory indicate that the natural physiological changes which occur in hormone levels, in particular changes in progesterone during proestrus, exert a substantial influence upon this extra-hypothalamic central nervous system site.     

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.     

Aging alters norepinephrine release in the medial preoptic area in response to steroid priming in ovariectomized rats

Changes in luteinizing hormone (LH) secretion that are observed in aging animals have been attributed to a reduction in hypothalamic norepinephrine (NE). The reason for the reduction in NE levels with aging is unclear. We hypothesized that the responsiveness of noradrenergic neurons to ovarian steroids is altered during aging. To test this, regularly cycling female Sprague-Dawley rats (young: 4-5 months old and middle age: 8-11 months old) were implanted with a push-pull cannula in the medial preoptic area (MPA) and ovariectomized bilaterally. On the 8th day after ovariectomy, they were injected with estrogen (30 microg/100 microl corn oil, s.c.) at 1000 h and on the 9th day they were implanted with a jugular catheter. On the 10th day they were injected with progesterone (2 mg/100 microl corn oil, s.c.) at 1000 h and subjected to push-pull perfusion. Perfusate samples from the MPA were collected at the rate of 10 microl/min every 30 min from 1300 to 1800 h and blood samples (0.3 ml) were collected hourly. The perfusate samples were analyzed for NE and dopamine (DA) concentrations using high performance liquid chromatography with electrochemical detection and serum LH levels were determined by RIA. In young animals, NE release (mean+/-S.E., pg/min) was 4.0+/-1.1 pg/min at 1300 h and increased significantly (p<0.05) to 10.4+/-4.3 pg/min at 1500 h and remained elevated until 1600 h and then declined to 6.8+/-2.5 at 1730 h. In contrast, the increase in NE release occurred briefly in middle-aged animals and was delayed by an hour. LH patterns in both age groups followed the pattern in NE release. There was no change in the release of DA in both young and middle-aged animals. It is concluded that the altered responsiveness of noradrenergic neurons to steroid priming in middle-aged rats probably plays a critical role in the alterations seen in LH secretion in older animals.     

Altered hypothalamic dopamine depletion rate and LHRH content in noncyclic hamsters

Effect of light deprivation on hypothalamic dopamine (DA) depletion rate and luteinizing hormone releasing hormone (LHRH) content was evaluated in golden hamsters. Hamsters maintained in continuous darkness (CD) became acyclic after about 6 weeks while another group of hamsters maintained vaginal cyclicity under 20 lux light intensity for 14 hours a day. All hamsters were ovariectomized subsequently. Luteinizing hormone releasing hormone levels were significantly higher in the medial basal hypothalamus (MBH) and the serum gonadotropin levels were significantly lower in CD hamsters when compared with 20 lux group. However, gonadotropin release in response to exogenous LHRH was not significantly different in CD or 20 lux group of hamsters. Examination of hypothalamic catecholamine levels one hour after α-methyl-p-tyrosine injection indicated that the DA depletion was higher in CD animals. Results indicate that light deprivation probably enhances DA depletion rate in the MBH, suggesting that increased activity in dopaminergic neurons may inhibit the release of MBH LHRH in hamsters kept in continuous darkness.     

Locus coeruleus lesions decrease norepinephrine input into the medial preoptic area and medial basal hypothalamus and block the LH, FSH and prolactin preovulatory surge

The aim of this work was to study the role of the dorsal noradrenergic ascending pathway (DNAP), which originates in the locus coeruleus (LC) on the preovulatory surge of luteinizing hormone (LH) follicle-stimulating hormone (FSH) and prolactin (PRL) by producing bilateral electrolytic lesions (cathodal or anodal) in this nucleus. LC lesions were placed at 11.00 h on proestrus in female rats with regular 4-day estrous cycles. Intact rats, sham-operated as well as animals with missed lesions served as controls. In Experiment I, anodal current was applied and hourly blood samples were withdrawn (from 13.00 to 17.00 h) via a jugular catheter from conscious, freely moving rats for determination of plasma LH, FSH and PRL concentrations. In Expt. II, Expt. I was repeated using cathodal current and collecting blood samples hourly from 13.00 to 18.00 h. In both experiments the animals were sacrificed on the next morning when the occurrence of ovulation was checked. The medial septal area (MSA), medial preoptic area (MPOA), and medial basal hypothalamus (MBH) were dissected and assayed for norepinephrine (NE), dopamine (DA) and 5-hydroxyindoleacetic acid (5-HIAA) content. Experiment III was performed in order to test if a hormonal discharge occurred immediately after lesion placement. Blood samples were collected immediately before and 15, 30, 60 and 90 min postoperatively (from 11.00 to 12.30 h). Either anodal or cathodal lesions blocked the proestrous surge of LH, FSH and PRL. The hypothesis that the lesions advanced or delayed these hormonal surges was rejected since we found no increases in the hormonal levels from 11.00 to 12.30 or from 13.00 to 18.00 h, and ovulation was not observed on the following morning in the lesioned animals. Since control, sham-operated and missed-lesion groups exhibited LH, FSH and PRL surges and ovulation, this blockage appears to be caused by the destruction of the LC neurons. Also, this blockade was correlated with a decrease in the NA content in the MPOA and MBH, but not in the MSA, whereas the DA and 5-HIAA content were not changed in all groups examined. The results lead us to suggest that the integrity of noradrenergic afferent input from the LC to luteinizing hormone-releasing hormone neurons in the MPOA and MBH is essential for triggering the preovulatory surge mechanisms for gonadotrophins and PRL.     

Striatal dopamine release is altered by glucose and insulin during push-pull perfusion of the rat's caudate nucleus

An individual site in the caudate-putamen complex of the unrestrained, fasted rat was tritium-labeled with 1.0 μCi of either³H-DA or ³H-NE. The amine was microinjected in a volume of 0.5 μl through a permanently implanted 20 ga guide cannula. Then 30 min later, a push-pull cannula assembly was lowered through the guide and the labeled site was perfused with an artificial CSF solution at a rate of 25 μl/min. Each perfusion lasted for 5 min with a 10 min interval between successive perfusions. At the mid-point of a sequence of seven perfusions, one of two experimental procedures was introduced: (1) the rat was offered food pellets which were readily consumed; or (2) one of three compounds was added to the CSF perfusate: (a) 55 μg/gml of D-glucose; (b) 4 mU/μl of insulin; or (c) 10 μg/μl of 2-DG. The results showed that the efflux of³H-DA increases as the rat feeds and during the subsequent perfusions. An immediate rise occurs in the output of the amine also when insulin is added to the perfusate; conversely, a slight suppression in ³H output is caused by glucose during its perfusion but a consistent elevation of radioactivity follows thereafter. 2-DG has no effect on the kinetics of catecholamine activity in the caudate-putamen. The patterns of release of ³H-NE are similar to those of ³H-DA under the same experimental conditions; however, the magnitude of the change is substantially attenuated. Although catecholamine activity within the caudate-putamen is usually implicated in motor control, our results support the possibility that dopamine in this structure may play a functional role in the regulation of food intake.     

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.     

Further Studies on the Suppression of Luteinizing Hormone Release Due to Activation of Medial Preoptic-Anterior Hypothalamic Area μ-Opioid Receptors

Our laboratory has previously demonstrated that specific activation of μ-opioid receptors in the medial preoptic-anterior hypothalamic area with [D-Ala², N Me-Phe⁴, Gly-ol⁵]-enkephalin (DAGO) suppresses luteinizing hormone secretion in the ovariectomized rat [29]. In the present study, three experiments were undertaken to ascertain whether changes in the activity of norepinephrine or dopamine neurons modulate the decrease in luteinizing hormone release in response to DAGO. The first experiment utilized push-pull perfusion in conjunction with HPLC to assess in vivo norepinephrine release in the medial preoptic-anterior hypothalamic area in response to perfusion of this site with DAGO (5 μg/h). DAGO significantly decreased luteinizing hormone release, but perfusate norepinephrine levels did not change. In the second experiment, push-pull perfusion in the medial preoptic-anterior hypothalamic area with cerebrospinal fluid (CSF) or CSF containing DAGO was done in rats pretreated with the norepinephrine synthesis inhibitor, FLA-63. This drug pretreatment had no effect on the DAGO-induced suppression of luteinizing hormone secretion. In Experiment 3, push-pull perfusion in the medial preoptic-anterior hypothalamic area with CSF followed by CSF containing DAGO was done in rats pretreated with vehicle, or a dopamine receptor antagonist, either pimozide or d-butaclamol. Neither dopamine receptor antagonist had any effect on the DAGO-induced suppression of luteinizing hormone release. Thus, these studies do not support a role for alterations in the activity of norepinephrine or dopmaine neurons in mediating the suppression of luteinizing hormone release in response to activation of μ-opioid receptors in the medial preoptic-anterior hypothalamic area in the ovariectomized rat. Copyright © 1997 Elsevier Science Inc.     

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.     

Starvation reduces norepinephrine activities in both hypothalamus and heart in rats

Norepinephrine (NE) activities in both hypothalamus and heart were simultaneously assessed in rats after food-deprivation for 2 days. The technique of gas chromatography-mass spectrometry was employed for the analysis of NE and its primary neuronal metabolite, 3,4-dehydroxyphenylethylene glycol (DHPG), and the ratio of DHPG to NE was used as an index of NE activity. Hypothalamic DHPG/NE ratio was significantly decreased by fasting and was completely reversed by a single day of refeeding. These changes in hypothalamic DHPG/NE ratio were parallel to those in cardiac DHPG/NE ratio. Supporting the concept in which hypothalamic NE neurons play an important role in modulating the sympathetic outflow, it is suggested the decrease in hypothalamic NE activity contributes to the reduction in cardiac NE activity during fasting.     

Differences in norepinephrine and dopamine neurotransmitter storage systems

Low doses of d-amphetamine (d-AMP) produced a 50% or greater decrease in the firing rates of both dopamine (DA) neurons (substantia nigra zone compacta) and norepinephrine (NE) neurons (locus coeruleus). However, pretreatment with the tyrosine hydroxylase inhibitor α-methyl-para-tyrosine (α-MT) blocked the d-AMP-induced reduction in DA neuron firing rate, but had no effect on the d-AMP-induced reduction in NE cell firing rate. Similarly, α-MT administered subsequent to d-AMP readily reversed the d-AMP-induced decrease in the firing rates of DA cells, but caused no significant reversal in NE cell firing rates. These electrophysiological findings, in conjunction with biochemical and behavioral data, support the hypothesis that there is a difference in the DA and NE neurotransmitter storage mechanism. In the DA neuron, there appears to be a slow transfer between stored and readily-releasable (newly synthesized) amine pools so that, following synthesis inhibition, there is little DA available for release. However, in the NE neuron, there is a more rapid mobilization of stored amine to readily releasable sites, such that d-AMP continues to cause the release of NE even though synthesis of transmitter is blocked.     

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.     

Brain catecholamine content during the estrous cycle and in steroid-primed rats

Norepinephrine (NE) and dopamine (DA) levels were measured in several estrogen concentrating brain regions over the estrous cycle and in steroid-primed ovariectomized rats under experimental conditions used to study sexual receptivity. Norepinephrine content in the ventromedial hypothalamus (VMH), lateral septum (LS) and medial preoptic area (POA) varied during the estrous cycle. The highest NE levels were found during metestrus in these brain regions. In the ovariectomized rat, NE levels were increased in the POA and LS 12 hours (hr) after the injection of 3 micrograms of estradiol benzoate (65% and 33%, respectively). Norepinephrine content in the POA and LS remained elevated 24 hr and 48 hr (0.5 mg progesterone at 42 hr) after treatment but were not different from control levels at 96 hr. Dopamine concentrations were not significantly altered over the estrous cycle or by the administration of estradiol benzoate in any of the brain regions studied. These results suggest that estrogen may selectively alter noradrenergic activity in brain regions that are implicated in the control of sexual receptivity and/or ovulation.     

Push-pull perfusion reveals meal-dependent changes in the release of bombesin-like peptides in the rat paraventricular nucleus

Bombesin (BN)-like peptides have been implicated in the regulation of ingestive behavior. The main objective of the present study was to monitor the dynamics of central BN-like peptide release in relationship to spontaneous meal ingestion and termination. Peptide level fluctuations were determined using in vivo push-pull perfusion of the hypothalamic paraventricular nucleus (PVN) and off PVN sites, combined with ex vivo radioimmunoassay. Analysis across all meals revealed significant differences between preprandial, prandial and postprandial extracellular BN-like immunoreactivity (BLI) at the anterior aspect of the PVN, with about a 3-fold diminution during a meal as compared to before or after a meal. Meal-related fluctuations were not detected at more distal hypothalamic sites or at sites within the caudate nucleus. When the analysis was restricted exclusively to the first meal after dark onset, a similar pattern of change in the interstitial levels of PVN BLI was generally observed; levels being higher preprandially as compared to the prandially (albeit by a smaller magnitude), and the termination of the first meal being accompanied by a robust (about 3-fold) increase in BLI. This is the first demonstration of site specific in vivo release of BN-like peptides in relation to feeding status and it further supports the physiological role of this family of peptides in the regulation of food intake.     

Effect of dopamine receptor blockade or norepinephrine synthesis inhibition on acute, ovariectomy-induced increases in pulsatile luteinizing hormone release in the rat

The initial aim of the present studies was to examine the influence of blockade of dopamine (DA) receptors with pimozide or inhibition of norepinephrine (NE) synthesis with U-14,624 on acute, ovariectomy (OVX)-induced changes in pulsatile LH release. Either treatment instituted at the time of OVX suppressed or inhibited the rapid increase in LH pulse amplitude and frequency normally occurring within 24 hr following ovarian removal on diestrus 1. While administration of pimozide at either 24 hr or 48 hr following OVX suppressed pulsatile LH release by selectively reducing LH pulse frequency, by 8 days following OVX pimozide failed to exert any effect on LH pulse frequency and therefore on pulsatile LH secretion. To determine if there was a transient critical period following OVX of at least 2 days but less than 8 when endogenous DA was excitatory to pulsatile LH release, piribedil (a DA receptor agonist) was given 24 hr following OVX. Rather than increase LH secretion, piribedil markedly suppressed pulsatile LH release indicating that DA does not stimulate LH secretion in acutely ovariectomized rats. These experiments indicate that

1. (1) NE is involved in stimulating the acute, OVX-induced increase that occurs in pulsatile LH release;

2. (2) DA receptorblockade by pimozide has a differential effect on pulsatile LH secretion which depends on the time following OVX when the compound is administered;

3. (3) this differential effect cannot be explained by a transient critical period of a few days duration following OVX during which DA is excitatory to pulsatile LH release.

Hypothalamic microinjection of norepinephrine (NE) elevates and prolongs basal efflux of unmetabolized NE from injection site

The effect of a microinjection of norepinephrine (NE) into the diencephalon of the rat was examined in terms of the subsequent endogenous activity of this amine. A push-pull guide cannula was first implanted stereotaxically with the tip resting in the rat's hypothalamus. Several days later, NE in a dose of 20 ng, 95 ng or 3.2 μg was microinjected in a volume of 0.5 μl into the perifornical region. After a 30 min interval, the injection site was perfused, by means of push-pull cannulae with an artificial CSF at a rate of 25 μl/min. The samples of perfusate collected one hour after the microinjection were analyzed by HPLC with electrochemical detection. The results indicate that an unexpectedly high level of unmetabolized NE can be detected at the perfusion site. This NE is greater than the picogram quantities of NE present endogenously in the rat's hypothalamus. The persistent presence of the exogenously applied amine within a microinjection site could explain the longevity of a given pharmacological effect of an amine often seen with this injection procedure. Also, it suggests that the local metabolic degradation of the amine is not as rapid as one would expect.     

Distribution of norepinephrine and dopamine in cerebral cortical areas of the rat

Concentrations of norepinephrine and dopamine were determined using enzyme isotope assay in 27 microdissected cerebral cortical areas of the rat. A detailed map is presented for microdissection of rat cerebral cortex. Norepinephrine was found in low but still measurable quantities throughout the cortex. Differences between cortical areas are also low. Relatively highest levels were demonstrated in the pyriform, insular and entorhinal cortices. The distribution of dopamine was found to be uneven with a maximal regional difference of 1:24. Concentration of dopamine was in all areas lower than that of norepinephrine. The highest dopamine concentration (2,4 ng/mg protein) was measured in the rostral pyriform cortex but other mesocortical (cingulate, frontal, insular and entorhinal) dopaminergic areas also contained relatively high amounts. Except for the caudal occipital and caudal entorhinal cortices all regions studied contained measurable quantities of dopamine. Its low concentration relative to norepinephrine (below 15%) suggests that in the cortical areas studied dopamine is present as the precursor of norepinephrine.     

Effects of dopamine and norepinephrine on neuronal activity of the olfactory tubercle

The effects of iontophoretic administration of norepinephrine (NE) and dopamine (DA) on olfactory tubercle (OT) neurons that respond to lateral hypothalamus (LH) or locus coeruleus (LC) electrical stimulation were studied. NE and DA decreased the frequency of OT neurons which were increased or decreased by the LH stimulation. An increased firing of OT neurons following NE or DA administration was less frequently observed. NE administration decreased the firing of OT neurons that responded to LC stimulation. These results suggest that the LC fibers which reach the OT use NE as a neurotransmitter. DA administration also suppressed the unitary discharge of OT neurons responding to LC stimulation. The increase in frequency of OT neurons observed following LH stimulation cannot be attributed to DA. The possibility that other suspected neural transmitters are involved in this effect is discussed.