Effects of different opiates on hypothalamic monoamine turnover and on plasma LH levels in pro-oestrous rats

Opiate inhibition of luteinizing hormone (LH) appears to involve changes in hypothalamic monoaminergic activity. Agonists of mu-, kappa- and sigma-opioid receptors and an opiate antagonist were administered at the onset of the preovulatory LH surge and their effects on hypothalamic monoamine turnover, and on plasma LH levels, investigated. The opiate antagonist, naloxone, significantly increased both noradrenaline (NA) turnover and plasma LH levels. Morphine (mu-agonist), significantly decreased NA concentration and plasma LH levels, but significantly increased dopamine (DA) and serotonin (5-HT) activity. Levorphanol (another mu-agonist) significantly decreased both NA and 5-HT concentrations and had no effect on circulating LH. Cyclazocine, ketocyclazocine and tifluadom (kappa-agonists) increased NA turnover but only tifluadom increased 5-HT turnover, also reducing LH levels significantly. N-Allylnormetazocine (SKF 10,047; sigma-agonist) increased 5-HT activity but did not alter LH levels. This study has confirmed the existence of a heterogenous group of opioid receptors within the hypothalamus which modulate monoamine neurotransmitters controlling LH release.     

Ontogeny of mu-, delta- and kappa-opioid receptors mediating inhibition of neurotransmitter release and adenylate cyclase activity in rat brain

The ontogeny was examined of functional opioid receptors mediating presynaptic inhibition of neurotransmitter release and inhibition of dopamine (DA)-sensitive adenylate cyclase in the rat brain, using highly selective agonists for mu-, delta- and kappa-receptors. On gestational day 17 (E17) strong inhibitory effects of the selective mu-agonist DAGO on the electrically evoked release of [3H]noradrenaline from cortical slices and of the selective kappa-agonist U-50,488 on the electrically evoked release of [3H]DA from striatal slices were found. Electrically evoked release of [3H]acetylcholine from striatal slices was not detectable before postnatal day 7 (P7), but on that day it was already strongly inhibited by the selective delta-agonist DPDPE. Although mu- and delta-opioid receptors coupled to DA-sensitive adenylate cyclase in the striatum are likely to be physically associated in an opioid receptor complex in the adult, they were found to develop asynchronously. Whereas selective activation of mu-receptors with DAGO resulted in an inhibition of D1 dopamine receptor-stimulated adenylate cyclase activity on E17, activation of delta-receptors with DPDPE was not effective until P14. This study confirms the early appearance of mu- and kappa-opioid receptors and the relatively late development of delta-opioid receptors in the rat brain. Most importantly, it shows that in an early stage of development opioids are already able to mediate modulation of noradrenergic (via activation of mu-receptors) and dopaminergic (via activation of mu- and kappa-receptors) neurotransmission processes. Therefore, these opioid receptor types could play a role in brain development and/or developmental disturbances.     

Kappa-Opioid Receptor Involvement in the Regulation of Pulsatile Luteinizing Hormone Release During Early Pregnancy in the Rat

The object of this study was to gain further insight into endogenous opioid peptide suppression of pulsatile luteinizing hormone (LH) release in early gestation in the rat by examining whether selective blockade of mu -, delta -, or kappa-opioid receptor(s) results in stimulation of pulsatile LH secretion at this time. Previous reports demonstrated stimulation of pulsatile LH release during early gestation by intravenous infusions of naloxone, an endogenous opioid peptide receptor antagonist whose binding is not specific to a single class of opioid peptide receptors. In the present study, naloxone infused intraventricularly similarly stimulated an increase in pulsatile LH release on Days 7 to 8 of gestation. Antagonists of specific opioid peptide receptor subtypes were thus given by this route. Administration of nor-binaltorphimine, an antagonist of kappa-opioid receptors, but not β-funaltrexamine or ICI 174, 864, antagonists of mu- and delta-opioid receptors, respectively, exerted a stimulatory action on both LH pulse amplitude and frequency similar to that of naloxone, indicating involvement of this opioid peptide receptor subtype in the endogenous opioid peptide suppression of pulsatile LH release in early gestation in the rat.     

Rostral ventrolateral medullary opioid receptor subtypes in the inhibitory effect of electroacupuncture on reflex autonomic response in cats

Electroacupuncture (EA) is used in traditional Chinese medicine to treat arrhythmias, hypertension and myocardial ischemia. Our previous work suggests that the inhibitory effect of EA on the pressor reflex induced by bradykinin (BK) applied to the gallbladder is due, in part, to the activation of opioid receptors, most likely located in the rostral ventrolateral medulla (rVLM). However, specific opioid receptor subtypes, and hence the neurotransmitters. responsible for this inhibition are unknown. Therefore, in anesthetized cats, BK (10 microg/ml) was applied to the gallbladder to induce transient reflex increases in arterial blood pressure (BP). EA (1-2 mA, 5 Hz, 0.5 ms pulses) was delivered through acupuncture needles inserted bilaterally into Neiguan and Jianshi acupoints on forelimbs, overlying the median nerves. EA attenuated the BK-induced pressor response by 39%. Opioid receptor subtype antagonists or agonists were microinjected unilaterally into the rVLM. The mu- and delta-receptor antagonists CTOP and ICI 174,864, respectively, significantly attenuated the EA-induced inhibition for at least 30 min. The K-receptor antagonist (nor-BNI) was less effective and was shorter acting. Like EA, microinjection of mu- and delta-opioid agonists, DAGO and DADLE, respectively, into the rVLM significantly decreased the pressor responses. In contrast, the kappa-opioid agonist, U50,488, failed to alter the BK-induced pressor response. We conclude that a significant portion of inhibition of the gallbladder pressor response by EA is related to activation of mu- and delta-opioid receptors in the rVLM. The endogenous neurotransmitters for mu- and delta-opioid receptors, beta-endorphins and enkephalins, in the rVLM, therefore appear to play a role in the EA-related modulation of cardiovascular reflex responses. Conversely, dynorphin is less likely to be involved in this response.     

Stimulation of mu- and delta-opioid receptors enhances phosphoinositide metabolism in mouse spinal cord: evidence for subtypes of delta-receptors.

The accumulation of inositol phosphates (IPs) induced by agonist-activated opioid receptors was analysed in mouse spinal cord slices pre-labelled with myo-[3H]inositol. Agonists showing selectivity to mu-opioid receptors, morphine and [D-Ala2,MePhe4, Gly(ol)5]enkephalin (DAMGO), promoted concentration-dependent increases in the formation of IPs. The activation of delta-opioid receptors by the selective agonists [D-Pen2,5]enkephalin (DPDPE) and [D-Ala2]deltorphin II produced similar increases in phosphoinositide (PI) metabolism. Pre-treatment of the slices with pertussis toxin (PTX) blocked the effect of opioid agonists on IP production. The involvement of Gi/Go-protein (guanine nucleotide-binding protein) classes in this opioid effect is therefore suggested. The activity of the opioid agonists was reduced by the opioid antagonists naltrexone and naloxone. The antagonist at delta1-receptors, 7-benzylidenenaltrexone (BNTX), exhibited greater potency than the antagonists at delta2-receptors, naltriben methanesulphonate (NTB) or naltrindrole 5'-isothiocyanate (NT II), in reducing the activating effect of DPDPE on phosphoinositide metabolism. Conversely, NTB and NT II were more potent antagonists of the activity of [D-Ala2]deltorphin II than BNTX. This work demonstrates the coupling of spinal mu- and delta-opioid receptors to phospholipase C and the generation of IPs. It also provides biochemical evidence for pharmacological subtypes of delta-opioid receptors in the activation of this signalling pathway.     

Alpha2-adrenoceptor-mediated restraint of norepinephrine synthesis, release, and turnover during immobilization in rats

Stress-related release of norepinephrine (NE) in the brain and periphery probably underlies several neuroendocrine and neurocirculatory responses. NE might influence its own synthesis, release, and turnover, by negative feedback regulation via alpha2-adrenoceptors. We examined central and peripheral noradrenergic function by measuring concentrations of NE, dihydroxyphenylglycol (DHPG), and dihydroxyphenylacetic acid (DOPAC) in hypothalamic paraventricular nucleus (PVN) microdialysate and arterial plasma simultaneously during immobilization (IMMO) in conscious rats. The alpha2-adrenoceptor antagonist yohimbine (YOH) was injected i.p. or perfused locally into the PVN via the microdialysis probe. The i.p. YOH increased plasma NE, epinephrine (EPI), DHPG, dihydroxyphenylalanine, and DOPAC levels by 4.3, 7.3, 2.5, 0.6 and 1.8-fold and PVN microdialysate NE, DHPG, and DOPAC by 1. 2, 0.6 and 0.5-fold. The i.p. YOH also enhanced effects of IMMO on plasma and microdialysate NE, DHPG, and DOPAC. YOH delivered via the PVN microdialysis probe did not affect microdialysate or plasma levels of the analytes at baseline and only slightly augmented microdialysate NE responses to IMMO. The results indicate that alpha2-adrenoceptors tonically restrain NE synthesis, release, and turnover in sympathetic nerves and limit IMMO-induced peripheral noradrenergic activation. In the PVN, alpha2-adrenoceptors do not appear to contribute to these processes tonically and exert relatively little restraint on IMMO-induced local noradrenergic activation.     

Interactions Between Opioid Peptides and Adrenaline-Containing Neurons Modulate Luteinizing Hormone Secretion in Male Rats

There is increasing evidence that the opioid inhibition of luteinizing hormone (LH) secretion is mediated, at least in part, by catecholaminergic mechanisms. This study determined the effects of selective manipulation of noradrenergic and adrenergic systems on the ability of opiate receptor blockade to induce the release of LH in adult male rats. Selective depletion of hypothalamic noradrenaline levels by 80% following 6-hydroxydopamine infusions into the central tegmental tract did not alter the 2- to 3-fold increase in serum LH levels following opiate receptor blockade with naloxone (2.5 mg/kg). In contrast, both selective depletion of hypothalamic adrenaline by prior treatment with the phenylethanolamine N-methyltransferase inhibitor, LY134046 (2 × 50 mg/kg) and non-selective depletion of all three catecholamines with α-methyl-p-tyrosine (250 mg/kg), abolished the naloxone-induced increase in LH. These results suggest that the inhibition of LH secretion by endogenous opioid peptides is influenced by catecholaminergic neurotransmission and further support the view that adrenaline rather than noradrenaline or dopamine is of importance in this context.     

Noradrenaline release in the medial preoptic area during the rat oestrous cycle: temporal relationship with plasma secretory surges of prolactin and luteinising hormone

During the rat oestrous cycle, the afternoon of pro-oestrous is characterised by preovulatory surges of luteinising hormone (LH) and prolactin. On the afternoon of oestrous, a secretory surge of prolactin has also been reported. Because the medial preoptic area (MPOA) is known to regulate prolactin and LH secretory surges and noradrenaline has been demonstrated to stimulate these hormones release, we evaluated whether noradrenaline release in the MPOA was temporally associated with plasma prolactin and LH surges in cycling rats. During the 4 days of oestrous cycle, noradrenaline concentrations were determined in microdialysates from the MPOA, collected at 30-min intervals from 10.30 h to 19.00 h. Plasma prolactin and LH levels were measured in blood samples withdrawn hourly from 14.00 h to 19.00 h on pro-oestrous and from 13.00 h to 18.00 h on the other days of the cycle. On the afternoons of both pro-oestrous and oestrous, noradrenaline levels increased at 14.00 h and remained elevated until 16.30 h. Conversely, they were low and constant throughout metoestrous and dioestrous. Correlating with noradrenaline release in the MPOA, plasma prolactin surges occurred during the afternoons of both pro-oestrous and oestrous. On pro-oestrous, the afternoon LH surge was also preceded by the increase in MPOA noradrenaline whereas, during oestrous, LH secretion was low and unaltered. A temporal association between noradrenaline release and prolactin secretion suggests that noradrenergic neurotransmission in the MPOA regulates prolactin surges in female rats. Moreover, our data also suggest that MPOA noradrenaline requires specific conditions to physiologically regulate LH secretion, which seems to occur during the afternoon of pro-oestrous.     

Dopamine depletion produces augmented behavioral responses to a mu-, but not a delta-opioid receptor agonist in the nucleus accumbens: lack of a role for receptor upregulation.

Microinjection of either mu- or delta-opioid agonists into the nucleus accumbens produces an increased locomotor activity, and when the dopaminergic innervation of the nucleus accumbens is bilaterally lesioned, the locomotor response to the microinjection of mixed mu- and delta-opioid agonists is augmented. To determine whether the lesion-induced augmentation to opioids is specific to mu- or delta-opioid receptor activation, dopamine innervation of the nucleus accumbens was lesioned with 6-hydroxydopamine (6-OHDA), and the motor stimulant response to intra-accumbens microinjection of the selective mu-opioid agonist, Tyr-D-Ala-Gly-mePhe-Gly-OH (DAMGO), was compared to that of the delta-opioid agonist, [D-penicillamine2,5]-enkephalin (DPDPE). The lesions caused a 95% depletion of tissue dopamine levels in the nucleus accumbens of the DAMGO-injected rats compared to sham-lesioned rats. Horizontal and vertical photocell counts were significantly increased in response to DAMGO in 6-OHDA-lesioned compared to the sham-lesioned rats. This behavioral augmentation was dose dependent and blocked by naloxone. In rats with similar accumbal dopamine depletions (94%), the locomotor response to DPDPE was not enhanced. The augmentation in the behavioral response to DAMGO was not associated with a change in the Bmax or Kd of [125I]DAMGO binding in nucleus accumbens homogenates from lesioned rats. Likewise, using quantitative receptor autoradiography, no difference between 6-OHDA- and sham-lesioned rats was observed in [125I]DAMGO or [125I]DPDPE binding. Therefore, the augmented behavioral response to opioids in the nucleus accumbens following dopamine depletion relies predominately on mu-opioid receptor stimulation. However, this augmentation is not mediated by an alteration in the number or affinity of these receptors.     

Dexamethasone blocking effects on mu- and delta-opioid-induced seizures involves kappa-opioid activity in the rabbit

Previous data indicate that intracerebroventricular administration of agonists for mu- and delta-opioid receptors induces limbic seizures in rats, but no data are reported in rabbits. We found that the mu- and delta-opioid peptides [D-Ala(2)-N,Me-Phe(4)-Gly(5)-ol]enkephalin (DAMGO), beta-endorphin and deltorphin II, induced EEG non-convulsive hippocampal seizures, and changes in hippocampal background EEG, physical parameters and overt behaviour after central administration. Dexamethasone pre-treatment prevented DAMGO-, deltorphin II- and beta-endorphin-induced seizures as well as changes in background EEG, physical parameters and overt behaviour induced by mu-opioid agonists. Dexamethasone antagonism on opioid action was blocked by pre-treatment with a protein synthesis inhibitor, cycloheximide or by the kappa-opioid antagonist nor-binaltorphimine. Our data suggest that dexamethasone influences opioid actions at mu- and delta-receptors via a protein synthesis mechanism involving kappa-opioid receptors.     

Alteration in hypothalamic neuropeptide Y (NPY) secretion may underlie female reproductive ageing: induction of steroid-induced luteinising hormone surge by NPY in ovariectomised aged rats

A large body of evidence suggests that a defect in the hypothalamic function may be the primary cause of reproductive ageing in female rats. We have previously shown that luteinising hormone (LH)-surge associated changes in hypothalamic neuropeptide Y (NPY) gene expression and median eminence (ME) NPY levels seen in young rats do not occur in middle-aged (MA) rats. The present study examined whether hypothalamic NPY release is altered during the steroid-induced LH surge in ovariectomised (OVX) MA rats, and whether exogenous NPY initiates steroid-induced LH surge in OVX old rats. In the first study, NPY release from the ME-arcuate nucleus, as assessed by the push-pull cannula technique, was significantly increased before and during the progesterone-induced LH surge in oestrogen (E(2))-primed ovariectomised young rats (2-3 months old). This antecedent increase in NPY release seen in young rats was not apparent in MA rats (11-13 months old) in association with a delayed and attenuated LH surge. In the second study, whereas progesterone failed to induce LH surges in E(2)-primed ovariectomised old rats (23-25 months old), intracerebroventricular NPY (0.1-0.5 microg) injections at 1100, 1200 and 13.00 h resulted in LH surge induction in E(2) + progesterone-primed ovariectomised old rats. Because increased hypothalamic NPY synthesis and release is obligatory for the preovulatory LH discharge in young rats, the present findings suggest that alteration in NPY release from the ME-arcuate nucleus contributes to the delayed and reduced LH surges in MA rats and may be involved in the subsequent loss of the LH surges in old rats.     

A role for central postsynaptic alpha 2-adrenoceptors in glucoregulation.

Central or peripheral administration of the alpha 2-adrenoceptor agonist clonidine causes marked hyperglycemia in the rat. It is not clear whether this effect is mediated within the brain at either pre- or postsynaptic alpha 2-adrenoceptors or whether it is due to peripheral alpha 2-agonist actions. We employed computerized mass spectrometry to measure noradrenaline (NA) and its primary neuronal metabolite 3,4-dihydroxyphenylglycol (DHPG) in the medial basal hypothalamus of rats treated acutely with clonidine, the alpha 2-antagonist yohimbine, the postganglionic noradrenergic blocker guanethidine and the neuroglycopenic agent 2-deoxy-D-glucose (2-DG). That clonidine's hyperglycemic effect was due, in part, to an action at central alpha 2-adrenoceptors was indicated by the ability of guanethidine to significantly inhibit the glucose response. Because of clonidine's inhibition of hypothalamic NA release (assessed by the DHPG/NA ratio), presumably by presynaptic agonism, these data indicated that postsynaptic receptor stimulation by clonidine was involved in activating glucose release. Yohimbine markedly increased the hypothalamic DHPG/NA ratio, reflecting presynaptic stimulation of NA release, but at the same time inhibited the hyperglycemic response due to 2-DG administration. This latter effect to block hyperglycemia is consistent with antagonism of postsynaptic alpha 2-adrenoceptors involved in mediating hepatic glucose output. These data indicate a major role for postsynaptic alpha 2-adrenoceptors in glucoregulation.     

Repeated electroconvulsive shock downregulates the opioid receptors in rat brain

Ten consecutive daily electroconvulsive shocks (ECSs), which produce maximal tonic and clonic convulsions, caused reductions of mu- and delta-opioid receptor binding in the hypothalamus, hippocampus and caudate nucleus, but not in the frontal cortex and brainstem. These changes of opioid receptor binding were not observed in rats receiving a single ECS. Scatchard analysis revealed that ECS-induced reduction of mu- and delta-receptor binding was due to a decrease in the binding sites but not to a change in the binding affinity. Time course studies showed that 7 days after the end of 10 consecutive daily ECSs, both mu- and delta-receptor binding remained lower than those of sham controls. However, the effects of ECS on the opioid receptor binding disappeared in 2-3 weeks. These observations are consistent with the hypothesis that ECS treatments increase the release of opioid peptides in certain brain regions which in turn down-regulate the opioid receptors.     

Enhanced responses to stress induced by fat-feeding in rats: relationship between hypothalamic noradrenaline and blood glucose

High-fat-feeding in rats has been reported to enhance stress reactions, as assessed by elevation of blood glucose and corticosterone levels. This study was designed to investigate the relationship between changes in blood glucose and hypothalamic neuronal noradrenaline activity (HNNA), as indexed by the ratio of dihydroxyphenylethynyleneglycol (DHPG) to noradrenaline (NA), following physiological stress in high-fat-fed rats. Two groups of adult male Wistar rats were fed isocaloric diets high in fat (59% of calories) or starch (70% of calories). After 3 weeks each of these groups was further subdivided into (a) control, (b) 2 min ambient temperature (20°C) swim or (c) 2 swim in ice-cold water. Animals were decapitated 20 min after commencing the swim; trunk blood and sample of medial basal hypothalamus were obtained. Computerized gas chromatography/mass spectrometry was used to measure hypothalamic DHPG and NA concentrations. There were no differences between fat- and starch-fed rats in basal levels of serum glucose, insulin or corticosterone and no differences in DHPG, NA or DHPG/NA ratio. Compared to starch-fed rats, ambients swim stress in the fat-fed group produced significantly larger serum glucose (P < 0.01), serum corticosterone (P < 0.05), DHPG (P < 0.05), and DHPG/NA (< 0.01) responses. Following cold swim stress similar differences between fat- and starch-fed animals were observed. In addition, serum insulin was found to be significantly suppressed in the fat-fed group (P < 0.05) following cold swim. Serum glucose, insulin and corticosterone levels correlated significantly with the DHPG/NA ratio across all groups (R = 0.54, P < 0.001; R = −0.36, P < 0.01;and, R = 0.53 P < 0.01, respectively). In conclusion, these studies provide further evidence for enhancement of stress responses in fat-fed rats and suggest these responses may be mediated through increased hypothalamic noradrenergic activity.     

Neonatal testosterone modifies LH secretion in the adult female rat by altering the opioid-noradrenergic interaction in the medial preoptic area

Noradrenergic-opioid interaction in the medial preoptic area (MPOA) was examined in ovariectomised adult rats which were oestrogen-treated and had been injected neonatally with either testosterone propionate (TP) or vehicle (oil). The first experiment involved electrical stimulation of the ventral noradrenergic tract (VNAT) in anaesthetised rats. Blood samples were collected before and after the stimulation to determine plasma levels of luteinising hormone (LH). Approximately half of the animals received naloxone i.v. 15 min before the onset of stimulation. In all groups, stimulation of VNAT elicited a significant increase in plasma LH concentration. However, pretreatment with naloxone in androgenised rats, but not in oil-treated animals, almost doubled the LH increment due to stimulation. Naloxone had no effect on plasma LH concentrations in unstimulated control rats. In the second experiment hypothalamic slices containing the MPOA were preincubated with [3H]noradrenaline [( 3H]NA) and then subjected to electrical field stimulation under the conditions of (a) no drug added and (b, c) morphine superfusion without and with naloxone. The opioid agonist morphine significantly reduced the net release of [3H]NA in normal and TP-treated female rats. Addition of equimolar naloxone reversed this effect in normal females, whereas in the androgenised group it not only reversed this effect but elicited a significant increase in [3H]NA release. From these data we conclude that (1) neonatal testosterone treatment alters noradrenergic-opioid interaction regulating LH secretion in adult females and (2) the site of this change may be the presynaptic opioid input to the noradrenergic terminals in the MPOA.     

Mu- and delta-opioid receptor antagonists reduce levodopa-induced dyskinesia in the MPTP-lesioned primate model of Parkinson's disease

Long-term treatment of Parkinson's disease with levodopa is complicated by the emergence of involuntary movements, known as levodopa-induced dyskinesia. It has been hypothesized that increased opioid transmission in striatal output pathways may be responsible for the generation of dyskinesia. In this study, we have investigated the effect of blockade of opioid peptide transmission on levodopa-induced dyskinesia in a primate model of Parkinson's disease-the MPTP-lesioned marmoset. Coadministration of nonselective and mu- or delta-subtype-selective opioid receptor antagonists with levodopa resulted in a significant decrease in dyskinesia. There was no attenuation of the anti-parkinsonian actions of levodopa. These data suggest that specific mu- or delta-opioid receptor antagonists might be applicable clinically in the treatment of levodopa-induced dyskinesia in Parkinson's disease.     

Pre- and postsynaptic inhibition by opioids in rat striatum.

The physiological role of opioid peptides in the rat striatum was sought by intracellular recording in vitro. Excitatory synaptic potentials (mediated by glutamate or aspartate) and inhibitory synaptic potentials (mediated by GABA) were isolated pharmacologically and/or by positioning the stimulation electrode over the external capsule. Opioid agonists and antagonists selective for mu-, delta-, and kappa-receptors were applied by superfusion. Two main actions of opioids were observed. First, mu- and delta-selective opioids presynaptically inhibited the excitatory postsynaptic potential, whereas only delta-selective opioids decreased the inhibitory synaptic potential. Second, a small subpopulation of cells (not medium spiny neurons) were hyperpolarized by delta-selective agonists. The results indicate that the main action of opioids on striatal neurons is presynaptic inhibition of the corticostriate excitatory synaptic input.     

Effects of the noradrenergic neurotoxin DSP4 on growth and the gonadal steroid- and PMSG-induced release of LH in the prepubertal female rat

We have examined the effect of DSP4 treatment on PMSG-induced ovulation. A marked attenuation of the stimulatory effects of PMSG (7.5 I.U.) by DSP4 was evidenced by the significantly lower number of corpora lutea present in the ovaries of those animals which ovulated compared to controls. In addition, ovarian weight was lower in the DSP4 group. In a further experiment, we examined the effect of DSP4 on the induction of an LH surge by progesterone (P) in estradiol benzoate (EB) primed rats. DSP4 administration 2 hours prior to P eliminated the LH surge seen in controls. In view of our previous observations that DSP4 can interact with opioid receptors, we attempted to block its inhibitory effect on PMSG and EB/P stimulations. Coinjection of naloxone, an opioid antagonist, only partially prevented the influence of DSP4. It seems likely, therefore, that opioid receptors are not involved in the inhibitory effects of DSP4 described here. In further experiments, we studied the effects of DSP4 on spontaneous sexual maturation in female rats. DSP4 was administered (50 mg/kg, IP) on either day 5, day 23, day 29, or both day 24 and day 26 of life. Growth was inhibited and vaginal opening (VO) was significantly delayed in all except the day 29 group. However, VO occurred at the same body weight as the controls. By the end of the experiment, hypothalamic noradrenaline levels were not significantly different between control and DSP4-treated animals. The lack of an effect of DSP4 on the progression to puberty may be due to sufficient recovery of the central noradrenergic systems during the time course of the experiments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Noradrenergic modulation of gonadotrophin‐inhibitory hormone gene expression in the brain of Japanese quail

Gonadotrophin‐inhibitory hormone (GnIH) is a hypothalamic neuropeptide that inhibits gonadotrophin synthesis and release in birds and mammals. In Japanese quail, GnIH neurones express the noradrenergic receptor and receive noradrenergic innervation. Treatment with noradrenaline (NA) stimulates GnIH release from diencephalic tissue blocks in vitro. However, the effects of NA on hypothalamic GnIH gene expression have not been determined. We investigated noradrenergic regulation of GnIH gene expression in the brain of male quail using the selective noradrenergic neurotoxin N‐(2‐chloroethyl)‐N‐ethyl‐2‐bromobenzylamine hydrochloride (DSP‐4). We first showed that DSP‐4 reduced the number of noradrenergic (dopamine‐β‐hydroxylase immunoreactive) cells in the locus coeruleus (LoC) and specifically lowered the NA concentration in the hypothalamus of male quail. Other monoamines, such as dopamine and serotonin, were not affected by drug treatment. DSP‐4 did not decrease the numbers of noradrenergic cells of the lateral tegmental cell group, nor the plasma NA concentration. Decreased hypothalamic NA levels after DSP‐4 treatment did not change GnIH gene expression in the brains of quail during their interaction with conspecifics. On the other hand, GnIH gene expression increased in the brains of quail socially isolated for 1 hour after DSP‐4 treatment. These results suggest that some noradrenergic neurones have inhibitory effects on GnIH gene expression of the hypothalamus in solitary quail.