Acute and chronic effects of desipramine and clorgyline on alpha(2)-adrenoceptors regulating noradrenergic transmission in the rat brain: a dual-probe microdialysis study

1. The effects of desipramine (3 mg kg(-1) i.p.) and clorgyline (1 mg kg(-1) i.p.) on extracellular noradrenaline (NA) in the locus coeruleus (LC) and cingulate cortex were assessed in freely-moving rats by dual-probe microdialysis. Functional activities of alpha(2)-adrenoceptors regulating NA release in the LC and cingulate cortex were determined by systemic (0.3 mg kg(-1) i.p.) or local (0.1 - 100 microM) clonidine administration. 2. Extracellular NA was increased in the LC and cingulate cortex following acute desipramine but not clorgyline treatment. Systemic clonidine decreased NA similarly in desipramine-, clorgyline-, and saline-treated animals, in both brain areas. 3. Long-term (twice daily, 14 days) but not short-term (twice daily, 7 days) desipramine, and long-term clorgyline (once daily, 21 days) treatments increased NA (3 fold) in cingulate cortex but not in the LC. Following long-term treatments, responses of NA to systemic clonidine were attenuated in the LC and cingulate cortex. 4. Clonidine perfusion by reverse dialysis into the cingulate cortex decreased local NA (-55 +/- 9%). The effect was attenuated by long-term desipramine (-31 +/- 9%) and clorgyline (-10 +/- 2%) treatments. 5. Clonidine perfusion by reverse dialysis into the LC decreased NA in the LC (-89 +/- 2%) and in cingulate cortex (-52 +/- 12%). This effect was attenuated in the LC following long-term desipramine (-72 +/- 4%) and clorgyline (-62 +/- 12%) treatments but it was not modified in the cingulate cortex (-57 +/- 10% and -68 +/- 6%, respectively). 6. These findings demonstrate that chronic desipramine or clorgyline treatments increase NA in noradrenergic terminal areas and desensitize alpha(2)-adrenoceptors modulating local NA release at somatodendritic and terminal levels. However, somatodendritic alpha(2)-adrenoceptors that control LC firing activity are not desensitized.     

N-[1-(2-benzo[b]thiophenyl)Cyclohexyl]- piperidine (BTCP) exerts cocaine-like actions on drug-maintained responding in rats.

The effects of N-[1-(2-benzo[b]thiophenyl)cyclohexyl]- piperidine (BTCP), a phencyclidine derivative that acts as a potent dopamine reuptake inhibitor, were examined on cocaine self-administration in rats. The effects of BTCP (0, 4, 8, 16, and 32 mg/kg, i.p.) on cocaine self-administration were tested against cocaine doses on both the ascending (0.0625 mg/infusion) and descending (0.25 mg/infusion) limb of the dose-response function. BTCP decreased self-administration of the 0.25-mg cocaine dose in a dose-dependent manner. A 16-mg/kg dose of BTCP that strongly suppressed self-administration of the 0.25-mg cocaine dose increased the intake at the 0.0625-mg dose of cocaine. Moreover, cocaine and BTCP pretreatments produced similar patterns of decreases in self-administration of cocaine on the descending limb of the dose-response function. The results suggest that BTCP has cocaine-like actions and produces a leftward shift of the dose-response curve for cocaine self-administration, indicating that the phencyclidine analog may substitute under certain conditions for the reinforcing effects of cocaine in self-administering rats.     

Activation of noradrenergic locus coeruleus neurons by clozapine and haloperidol: involvement of glutamatergic mechanisms

The major brain noradrenergic nucleus locus coeruleus (LC) has long been thought to be involved in states of alertness and cognitive processes. These functional characteristics make this nucleus interesting with regard to the signs of schizophrenia, especially the negative symptoms of the disease. In the present in-vivo electrophysiological study we analyse a putative interaction between endogenous kynurenic acid (KYNA) and the antipsychotic drugs clozapine and haloperidol on noradrenergic LC neurons. Previous studies have shown that systemically administered antipsychotic drugs increase the neuronal activity of LC noradrenaline (NA) neurons. In line with these findings, our results show that clozapine (1.25-10 mg/kg i.v.) and haloperidol (0.05-0.08 mg/kg i.v.) increased the firing rate of LC NA neurons in anaesthetized rats. Pretreatment with PNU 156561A (40 mg/kg i.v., 3 h), a potent inhibitor of kynurenine 3-hydroxylase, produced a 2-fold increase in rat brain KYNA levels. This treatment prevented the increase in firing rate of LC NA neurons induced by haloperidol (0.05-0.08 mg/kg i.v.) and clozapine in high doses (2.5-10 mg/kg i.v.). However, the excitatory action of the lowest dose of clozapine (1.25 mg/kg i.v.) was not abolished by elevated levels of brain KYNA. Furthermore, pretreatment with L-701,324 (4 mg/kg i.v.) a selective antagonist at the glycine site of the NMDA receptor prevented the excitatory effects of both clozapine and haloperidol. The present results suggest that the excitation of LC NA neurons by haloperidol and clozapine involves a glutamatergic component.     

Inhibition of 5-hydroxytryptamine reuptake by the antidepressant citalopram in the locus coeruleus modulates the rat brain noradrenergic transmission in vivo

The in vivo effect of the serotonin (5-HT) reuptake inhibitor antidepressant citalopram, administered in the locus coeruleus (LC), on noradrenergic transmission was evaluated in the rat brain. In dual-probe microdialysis assays, citalopram (0.1-100 microM), in a concentration-dependent manner, increased extracellular noradrenaline (NA) in the LC and simultaneously decreased extracellular NA in the cingulate cortex (Cg). These effects of citalopram were abolished by pretreatment with the 5-HT synthesis inhibitor p-chlorophenylalanine (400 mg/kg, i.p.). When the alpha(2)-adrenoceptor antagonist RS79948 (1 microM) was perfused in the LC, local citalopram increased NA dialysate in the LC but no longer modified NA dialysate in the Cg. In electrophysiological experiments, the administration of citalopram (100 microM) in the LC by reversal dialysis, decreased the firing rate of LC neurones. The results demonstrate in vivo that local administration of citalopram in the LC leads to a decreased release of NA in the Cg. This modulation seems to be the result of an increase in NA concentration in the LC and the subsequent inhibition of LC neurones via alpha(2)-adrenoceptors. The effects of citalopram are dependent on the presence of endogenous 5-HT in the LC.     

Priming with BTCP, a dopamine reuptake blocker, reinstates cocaine-seeking and enhances cocaine cue-induced reinstatement

N-[1-(2-benzo[b]thiophenyl)cyclohexyl]piperidine (BTCP), a potent dopamine reuptake inhibitor, substitutes for the reinforcing effects of cocaine and meets other criteria for possible agonist pharmacotherapeutic potential. The purpose of this study was to determine (1) whether BTCP modifies reinstatement of cocaine-seeking elicited by cocaine-related environmental stimuli and (2) whether this compound produces priming effects. Male Wistar rats were trained to associate discriminative stimuli (S(D)) with cocaine availability (0.25 mg/infusion) versus non-reward and then were subjected to repeated extinction sessions during which the reinforcer and S(D) were withheld. Subsequent presentation of the cocaine S(D) produced recovery of cocaine-seeking. BTCP (2.5-30 mg/kg; i.p.) did not attenuate the conditioned reinstatement induced by the cocaine S(D) but, rather, potentiated this effect at 10 mg/kg. To test whether BTCP, by itself, exerts priming effects, different groups of rats were trained to self-administer cocaine (0.25 mg/infusion) for 2 weeks. After a 2-week extinction period, BTCP (5, 10 and 20 mg/kg, i.p.) reinstated cocaine-seeking, showing that BTCP not only increases cocaine-seeking induced by cocaine-related stimuli but also produces priming effects following abstinence. The results suggest that, in cocaine abstinent rats, BTCP produces cocaine-like effects.     

The effects of the phencyclidine analogs BTCP and TCP on nigrostriatal dopamine neuronal activity

Extracellular single unit recording techniques were used to evaluate the effects of two phencyclidine (PCP) derivatives. N-[1-(2-benzo(b)thiophenyl)cyclohexyl]piperidine (BTCP) and N-[1-(2-thiophenyl)cyclohexyl]piperidine (TCP) on the electrophysiological activity of antidromically identified nigrostriatal dopamine (DA) neurons in chloral hydrate-anesthetized rats. I.v. BTCP produced a dose-dependent decrease in the firing rate of identified nigrostriatal DA neurons whereas TCP elicited a dose-dependent biphasic effect which was characterized by an activation of cell firing at low doses followed by a reversal of the response with larger doses. A hemitransection of the brain anterior to the substantia nigra significantly reduced the inhibitory effect of BTCP while this surgical procedure did not affect the response to TCP. However, iontophoretic application of BTCP induced a current-dependent inhibition of the spontaneous activity of cells while local application of TCP had no effect on the firing rate of these cells. These data indicate that PCP analogs are able to interact with the nigrostriatal DAergic pathway through distinct and opposing mechanisms. The results are discussed in light of recent observations that BTCP is selective for the DA uptake site while TCP is selective for the high affinity PCP binding site.     

Noradrenaline release in rat locus coeruleus is regulated by both opioid and alpha(2) -adrenoceptors.

Extracellular fluid levels of noradrenaline (NA) in the locus coeruleus (LC) during naloxone-precipitated morphine withdrawal with pretreatment of yohimbine (1 mg kg (-1), s.c.) or clonidine (1 mg kg (-1), s.c.) were measured in rats. There was a significant increase in the NA level after the injection of naloxone (2 mg kg (-1), i.p.) in the morphine-dependent rats. Moreover, the NA levels in the LC markedly increased during the 30-60 min following the naloxone (i.p.) challenge in the morphine-dependent rats pretreated with yohimbine. In contrast, the naloxone challenge in morphine-dependent rats pretreated with clonidine notably decreased the levels of NA in the LC. Behavioral signs of withdrawal were observed following the naloxone challenge in the morphine-dependent rats pretreated with yohimbine, with minimal signs in the morphine-infused rats pretreated with clonidine, and none in the saline-infused controls. These results directly suggest that NA increased within the LC after the naloxone challenge in morphine-dependent animals pretreated with yohimbine may be, at least in part, regulated by alpha(2) -adrenoceptors in the LC.     

Further characterization of brain 3,4-dihydroxyphenylethyleneglycol (DHPG) formation: dependence on noradrenergic activity and site of formation

The dependence of brain and plasma 3,4-dihydroxyphenylethyleneglycol (DHPG) formation upon CNS noradrenergic neutronal activity was evaluated following manipulations that are known to alter the firing rate of the locus coeruleus (LC) neurons and as a consequence, noradrenaline (NA) release and turnover. In addition, the relative degree of intraneuronal formation of brain DHPG was assessed by studying the metabolism of released NA during uptake inhibition. Electrical stimulation of the LC for 20 min induced an increase in rat cortical (40-42%), hypothalamic (22-29%) and plasma (68-79%) total DHPG and 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG) levels. Two hours following administration of the alpha-2 adrenoceptor antagonist yohimbine (10 mg/kg, i.p.), rat brain cortical conjugated DHPG and MHPG as well as free MHPG concentrations were increased whereas cortical free DHPG levels remained unchanged. The same treatment also increased plasma total DHPG and MHPG levels. In mice given the NA uptake inhibitor desipramine (10 mg/kg, i.p.) 2 h prior to sacrifice, brain free DHPG and MHPG concentrations were significantly reduced by 30 and 40%, respectively, whereas yohimbine (1-20 mg/kg, i.p.) induced a dose-dependent increase in brain DHPG (60-80%) and MHPG (60-220%) concentrations. Pretreatment with desipramine (10 mg/kg, i.p.) 30 min prior to yohimbine reduced, in rat, or abolished, in mice, the yohimbine-induced elevation of brain DHPG levels. In contrast, desipramine augmented the effect of yohimbine on brain MHPG levels resulting in a shift to the left of the dose response curves. These findings indicate that brain and plasma DHPG levels are sensitive to changes in brain noradrenergic neuronal impulse flow.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pharmacological characterization of the discriminative stimulus properties of the phencyclidine analog, N-[1-(2-benzo(b)thiophenyl)-cyclohexyl]piperidine

Rationale: Although both cocaine and the phencyclidine analog, BTCP, have dopamine (DA) re-uptake blocking properties, under some conditions their behavioral effects can be differentiated. Therefore, we examined whether the discriminative stimulus (DS) effects of BTCP are different from those of cocaine. Objectives: To compare the effects of monoamine re-uptake blockers, varying in their in vitro potencies as inhibitors of DA, norepinephrine (NE), or serotonin re-uptake, in different groups of rats trained to discriminate either BTCP or cocaine from saline. Additionally, drugs from other pharmacological classes were tested in both groups. Methods: Rats were trained to discriminate either BTCP (5 mg/kg, i.p.) or cocaine (10 mg/kg, i.p.) from saline under a two-lever FR10 drug discrimination procedure. Results: BTCP and cocaine cross-substituted in BTCP- and cocaine-trained rats. The DA re-uptake blockers, mazindol, indatraline, methylphenidate, GBR12909, and GBR12935, occasioned dose-related drug-lever (DL) selection both in cocaine- and in BTCP-trained rats, with potencies that were significantly correlated. In contrast, the NE re-uptake blockers, nisoxetine, desipramine, and nortriptyline, produced higher levels of DL selection in BTCP-trained rats than in cocaine-trained rats, a profile like that reported in low-dose cocaine-trained rats. Drugs from other classes acted similarly in both discriminations. Further, the α₁-adrenergic antagonist prazosin dose dependently blocked the DS effects of the training dose of BTCP, but not of cocaine. Conclusions: Theresults suggest that the DS effects of BTCP are similar to cocaine, and resemble those of a low training dose of cocaine. Received: 6 October 1998 / Final version: 19 March 1999     

1-[1-(2-Benzo[b]thiopheneyl)cyclohexyl]piperidine hydrochloride (BTCP) yields two active primary metabolites in vivo. Identification and quantification of BTCP primary metabolites in mice plasma, urine, and brain and their affinity for the neuronal dopamine transporter.

1-[1-(2-Benzo[b]thiopheneyl)cyclohexyl]piperidine hydrochloride (BTCP) and cocaine bind to the neuronal dopamine transporter (DAT) to strongly inhibit dopamine (DA) reuptake. Although similar to acute administration, cocaine and BTCP produce sensitization and tolerance, respectively, on chronic administration. We previously found that liver microsomes produced two primary metabolites from BTCP with a high affinity for DAT. Because such metabolites, if produced in vivo, could account for the pharmacological difference with cocaine, it was important to compare BTCP biotransformations in vitro and in vivo. Therefore, we identified and quantified BTCP and primary metabolites in mice urine, plasma, and brain after acute i.p. administration. The low recovery yield suggest that BTCP might behave like its close analogue, phencyclidine, with long-term storage of metabolites. Two active metabolites found in vitro were found in mice brain with estimated half-life values similar to that of BTCP (0.3 h). Although respective brain concentrations were 20 and 40 times lower than that of BTCP, their potency to displace in vivo [³H]BTCP bound to the DAT was 50 and 10 times higher, respectively, than that of BTCP. They could, therefore, contribute to the inhibition of DA transport and play an important role in BTCP pharmacology. They could also explain the differences between BTCP and cocaine on repeated administration.     

Comparative responses of the central adrenaline- and noradrenaline-containing neurons after reserpine injections

The responses of the noradrenaline (NA)- and adrenaline (A)-containing neurons to a reserpine treatment have been studied in the rat brain by using biochemical indices of the neuronal activity. Three days after multiple reserpine injections, tyrosine hydroxylase activity was significantly increased in the locus coeruleus (LC), A1-C1 and C2 regions. No change in this activity was observed in the A2 region. Furthermore, the NA and A endogenous levels were markedly reduced both in NA and A cell bodies and/or terminals, suggesting a reserpine action on NA and A neurons. The NA turnover was unchanged in all the regions analyzed. Conversely, the A turnover was reduced in the LC, A2 and C2 regions and in the nucleus periventricularis of the hypothalamus. This result suggests a different degree of sensitivity and/or response of the NA and A neurons following reserpine administration.     

1-[1-(2-Benzo[b]thiopheneyl)cyclohexyl]piperidine hydrochloride (BTCP) yields two active primary metabolites in vivo: Identification and quantification of BTCP primary metabolites in mice plasma, urine, and brain and their affinity for the neuronal dopamine transporter

1-[1-(2-Benzo[b]thiopheneyl)cyclohexyl]piperidine hydrochloride (BTCP) and cocaine bind to the neuronal dopamine transporter (DAT) to strongly inhibit dopamine (DA) reuptake. Although similar to acute administration, cocaine and BTCP produce sensitization and tolerance, respectively, on chronic administration. We previously found that liver microsomes produced two primary metabolites from BTCP with a high affinity for DAT. Because such metabolites, if produced in vivo, could account for the pharmacological difference with cocaine, it was important to compare BTCP biotransformations in vitro and in vivo. Therefore, we identified and quantified BTCP and primary metabolites in mice urine, plasma, and brain after acute i.p. administration. The low recovery yield suggest that BTCP might behave like its close analogue, phencyclidine, with long-term storage of metabolites. Two active metabolites found in vitro were found in mice brain with estimated half-life values similar to that of BTCP (0.3 h). Although respective brain concentrations were 20 and 40 times lower than that of BTCP, their potency to displace in vivo [³H]BTCP bound to the DAT was 50 and 10 times higher, respectively, than that of BTCP. They could, therefore, contribute to the inhibition of DA transport and play an important role in BTCP pharmacology. They could also explain the differences between BTCP and cocaine on repeated administration.     

BTCP is a potent reinforcer in rats: comparison of behavior maintained on fixed- and progressive-ratio schedules

N-[1-(2-benzo[b]thiophenyl)cyclohexyl]piperidine (BTCP) is a phencyclidine (PCP) derivative that acts as a potent dopamine (DA) reuptake inhibitor. Earlier studies have shown that BTCP can substitute for the reinforcing effects of cocaine. Therefore, the aim of the study was to further characterize the reinforcing effects of BTCP. The reinforcing actions of BTCP were compared to those of cocaine at equimolar concentrations in drug-na?ve rats. Two groups of animals were implanted with jugular catheters and trained to intravenously self-administer BTCP or cocaine (0.25 mg/infusion) on a fixed-ratio five schedule (FR 5) of reinforcement. Both BTCP and cocaine produced comparable inverted U-shaped dose-effect curves on this schedule over doses of 0.03, 0.06, 0.125, and 0.25 mg/infusion. Two doses (0.125 and 0.25 mg/infusion) that produced reliable self-administration in all the animals for cocaine and BTCP were then tested on a progressive-ratio schedule. At each dose, BTCP supported higher breaking points (BPs) than cocaine. The results demonstrate that rats readily acquire responding maintained by BTCP and suggest that BTCP may have greater reinforcing effects than cocaine at equimolar concentrations.     

Control of 5-hydroxytryptamine release in the dorsal raphe nucleus by the noradrenergic system in rat brain. Role of alpha-adrenoceptors.

The interactions between the brainstem serotonergic (5-hydroxytryptamine, 5-HT) and noradrenergic (NA) systems are important for the pathophysiology and treatment of affective disorders. We examined the influence of alpha-adrenoceptors on 5-HT and NA release in the rat dorsal raphe nucleus (DR) using microdialysis. 5-HT and NA concentrations in DR dialysates were virtually suppressed by TTX and increased by veratridine. The local and systemic administration of the alpha(1)-adrenoceptor antagonist prazosin reduced the DR 5-HT output but not that of NA. The maximal 5-HT reduction induced by local prazosin administration (-78% at 100 microM) was more marked than by its systemic administration (-43% at 0.3 mg/kg). The local application of NA and desipramine, to increase the tone on DR alpha(1)-adrenoceptors, did not enhance 5-HT release. The local (100 microM) or systemic (0.1-1 mg/kg s.c.) administration of clonidine reduced 5-HT and NA release (-48 and -79%, respectively, at 1 mg/kg), an effect reversed by RX-821002, which by itself increased both amines when given systemically. DSP-4 pretreatment prevented the effects of clonidine on 5-HT, suggesting the participation of alpha(2)-adrenoceptors on NA elements. Moreover, the systemic effect of clonidine on 5-HT (but not NA) was cancelled by lesion of the lateral habenula and by anesthesia, and was slightly enhanced by cortical transection. These data support the view that alpha(1)-adrenoceptors in the DR tonically stimulate 5-HT release, possibly at nearly maximal tone. Likewise, the 5-HT release is modulated by alpha(2)-adrenoceptors in NA neurons and in forebrain areas involved in the distal control of 5-HT neurons.     

Nociceptin, Phe(1)psi-nociceptin(1 - 13), nocistatin and prepronociceptin(154 - 181) effects on calcium channel currents and a potassium current in rat locus coeruleus in vitro

1. The actions of the neuropeptide nociceptin, the putative nociceptin receptor antagonist [Phe1psi(CH(2)-NH)Gly(2)]-nociceptin-(1 - 13)NH(2) (Phe(1)psi-nociceptin(1 - 13)) and the putative nociceptin precursor products nocistatin (rat prepronociceptin(125 - 132)) and rat prepronociceptin(154 - 181) were examined on membrane properties of rat locus coeruleus (LC) neurons using whole cell patch clamp techniques. 2. Nociceptin inhibited I(Ba) in all LC neurons, (pD(2) of 8.9, maximum inhibition 50%). The inhibition of I(Ba) by nociceptin was associated with slowing of the activation of I(Ba) and could be significantly reversed by a strong depolarizing prepulse. Phe(1)psi-nociceptin(1 - 13) also inhibited I(Ba) in LC neurons (notional pD(2) of 7.6, maximum inhibition 18%). Application of Phe(1)psi-nociceptin(1 - 13) (1 microM) significantly occluded the subsequent effects of a co-application of nociceptin (3 nM) on I(Ba). 3. As previously reported for nociceptin, Phe(1)psi-nociceptin(1 - 13) caused an outward current in LC neurons voltage clamped at -60 mV (pD(2) of 7.1, maximum current 50% of that of methionine enkephalin, 10 microM). The Phe(1)psi-nociceptin(1 - 13) induced current reversed polarity at -112 mV and exhibited pronounced inward rectification. Phe(1)psi-nociceptin(1 - 13) (1 microM) reversibly inhibited the current caused by nociceptin (300 nM) by 30%. 4. Neither nocistatin nor rat prepronociceptin(154 - 181) inhibited I(Ba) in LC neurons, or prevented the subsequent inhibition by nociceptin. Neither nocistatin or prepronociceptin(154 - 181) affected the membrane properties of LC neurons. 5. This study demonstrates that nociceptin modulates somatic I(Ba) in rat LC neurons. The putative ORL1 antagonist Phe(1)psi-nociceptin(1 - 13) exhibited partial agonist activity at inhibiting I(Ba) and opening K(+) channels in LC. Other putative nociceptin precursor products were without effect on LC cells.     

Electrophysiological effects of cocaine on central monoaminergic neurons

The electrical activity of three different single, identified, spontaneously firing central neurons was monitored by extracellular microelectrodes. Intravenous cocaine administration (0.25-2 mg/kg) elicited an activation of cerebellar Purkinje neurons (PN) and an inhibition of serotonergic dorsal raphe (DRN) and noradrenergic locus coeruleus (LC) neurons. These effects were not temporally correlated with the increase in mean arterial pressure elicited by the intravenous administration of cocaine. In addition, the administration of procaine, a structurally related local anesthetic, did not significantly affect any of the three central neurons studied. Our results suggest that cocaine has potent effects on the activity of DRN, LC and PN neurons, which are not directly related to its cardiovascular or local anesthetic actions.     

Non reciprocal cross-sensitization between cocaine and BTCP on locomotor activity in the rat

Measurement of locomotor sensitization was employed to characterize the effect of intermittent treatment with N-[1-(2-benzo[b]thiophenyl)cyclohexyl]piperidine (BTCP) and cocaine in the rat. Like cocaine, BTCP possesses high affinity for the dopamine transporter and inhibits dopamine reuptake. Although both drugs exhibit similar behavioral and neurochemical profiles with acute administration, there is tentative evidence to suggest that following chronic treatment BTCP does not induce neurochemical sensitization, and can attenuate cocaine-induced neurochemical sensitization in the striatum. Male Wistar rats were randomly divided into five groups after determining baseline locomotor activity. Three groups were treated with either saline (saline/saline), cocaine (20 mg/kg; cocaine/cocaine), or BTCP (10 mg/kg; BTCP/BTCP) for 10 days. The remaining two groups were treated with cocaine (20 mg/kg) or BTCP (10 mg/kg) for 3 days, followed by administration of BTCP (10 mg/kg; cocaine/BTCP) or cocaine (20 mg/kg; BTCP/cocaine) for 7 days. Locomotor sensitization was observed in all groups. However, although cross-sensitization on the day of substitution (day 4) was found in the BTCP/cocaine group, cross-sensitization was not observed in the cocaine/BTCP group. These results suggest that although the locomotor-activating effects of BTCP and cocaine are similar, the two drugs do not act identically, and different neural mechanisms may underlie BTCP and cocaineinduced sensitization.     

No apparent difference in the effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on the sympathetic system in NMRI and C57 BL/6 mice

The parkinsonism-inducing neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) has been shown to possess marked species as well as strain differences in toxicity on central catecholaminergic systems. In the present study the effects on the peripheral sympathetic nervous system following treatment with MPTP, as well as its metabolite 1-methyl-4-phenylpyridine (MPP+) and the catecholamine neurotoxin 6-hydroxydopamine (6-OHDA) were studied in mice of the NMRI and C57 BL/6 strains, two strains that possess marked difference in MPTP toxicity on central catecholaminergic neurons. No strain differences in the depletions of noradrenaline (NA) in iris and heart auricula and of NA and dopamine (DA) in superior cervical ganglion or in the reduction of the in vitro [3H]NA uptake in iris or heart auricula were found following MPTP treatment (2 X 40 mg/kg s.c., 2 and 7 days). Treatment with the NA uptake blocker desipramine (DMI) did not affect the MPTP-induced NA depletion in either strain. Following treatment with MPP+ (30 mg/kg i.v., 7 days) no differences in the two strains were seen on the reduction of NA levels in iris and heart auricula or decrease in [3H]NA uptake. In addition, no differences were found on NA levels in iris and heart auricula after 6-OHDA treatment (15 mg/kg i.v., 7 days). The data indicate that in the NMRI and C57 BL/6 mice peripheral NA neurons do not possess any notable strain difference in the vulnerability to MPTP or in the mechanism of action of MPTP.     

Regulation of central noradrenergic activity by 5-HT(3) receptors located in the locus coeruleus of the rat

A functional interaction between serotonergic and noradrenergic systems has been shown in the locus coeruleus (LC). Noradrenaline (NA) levels in the prefrontal cortex (PFC) are dependent on the firing rate of LC neurons, which is controlled by α₂ adrenoceptors (α2ADR). The aim of the present study was to investigate the role of 5-HT₃ receptors (5HT3R) in the modulation of central noradrenergic activity. We measured extracellular NA concentrations in the LC and PFC by dual-probe microdialysis in awake rats and the firing rate of LC neurons by electrophysiological techniques in vitro. Administration of the 5HT3R agonists SR57227 (1–100 μM) and m-chlorophenylbiguanide (mCPBG, 1–100 μM) into the LC increased NA in this nucleus (E_max = 675 ± 121% and E_max = 5575 ± 1371%, respectively) and decreased NA in the PFC (E_max = ?49 ± 6% and E_max = ?25 ± 11%, respectively). Administration of the 5HT3R antagonist Y25130 (50 μM) into LC attenuated SR57227 effect in the LC (E_max = 323 ± 28%) and PFC (E_max = ?37 ± 7%). The α2ADR antagonist RS79948 (1 μM) blocked the SR57227 effect in the PFC but it did not change the effect in the LC (E_max = 677 ± 202%). In electrophysiological assays, both mCPBG (1–10 μM) and SR57227 (1–10 μM) reduced the firing rate of about 50% of tested LC neurons (maximal effect = ?37 ± 2% and ?31 ± 4%, respectively); this effect was partially blocked by Y25130 (50 μM). Administration of RS79948 (1 μM) reversed the inhibition induced by mCPBG. Competition radioligand assays against [³H]UK14304 and [³H]RX821002 (α2ADR selective drugs) in the rat brain cortex showed a very weak affinity of SR57227 for α2ADR, whereas the affinity of mCPBG for α2ADR was 17-fold higher than that of SR57227 for α2ADR. The present results suggest that 5HT3R stimulate NA release in the LC, which promotes simultaneously a decrease in the firing rate of LC neurons through α2ADR and then a decrease of NA release in terminal areas such as the PFC.     

Characterization of noradrenaline release in the locus coeruleus of freely moving awake rats by in vivo microdialysis

Rationale The origin and regulation of noradrenaline (NA) in the locus coeruleus (LC) is unknown.Objectives The neurochemical features of NA overflow (nerve impulse dependence, neurotransmitter synthesis, vesicle storage, reuptake, ₂-adrenoceptor-mediated regulation) were characterized in the LC.Methods Brain microdialysis was performed in awake rats. Dialysates were analyzed for NA.Results NA in the LC decreased via local infusion of Ca²⁺-free medium (–42±5%) or the sodium channel blocker tetrodotoxine (TTX) (–47±8%) but increased (333±40%) via KCl-induced depolarization. The tyrosine hydroxylase (TH) inhibitor -methyl-p-tyrosine (250 mg kg^–1, i.p.) and the vesicle depletory drug reserpine (5 mg kg^–1, i.p.) decreased NA. Therefore, extracellular NA in the LC satisfies the criteria for an impulse flow-dependent vesicular exocytosis of neuronal origin. Local perfusion of the ₂-adrenoceptor agonist clonidine (0.1–100 M) decreased NA (E_max=–79±5%) in the LC, whereas the opposite effect (E_max=268±53%) was observed with the _2A-adrenoceptor antagonist BRL44408 (0.1–100 M). This suggests a tonic modulation of NA release through local _2A-adrenoceptors. The selective NA reuptake inhibitor desipramine (DMI) (0.1–100 M) administered into the LC increased NA in the LC (E_max=223±40%) and simultaneously decreased NA in the cingulate cortex, confirming the modulation exerted by NA in the LC on firing activity of noradrenergic cells and on the subsequent NA release in noradrenergic terminals.Conclusion Synaptic processes underlying NA release in the LC are similar to those in noradrenergic terminal areas. NA in the LC could represent local somatodendritic release, but also the presence of neurotransmitter release from collateral axon terminals.