Elevated BDNF mRNA expression in the medial prefrontal cortex after d-amphetamine reinstated conditioned place preference in rats.

Drug addiction behavior that is established and maintained by psychostimulants has been shown to be associated with the expression of brain-derived neurotrophic factor (BDNF) in the mesolimbic dopamine (DA) system. Cocaine has been used for most prior studies testing this effect of psychostimulants and therefore relatively little is known about its counterpart amphetamine (AMP). To fill this gap, the present study was designed to test whether BDNF mRNA expression levels in the DA terminal regions were changed specifically by d-AMP-induced conditioned place preference (CPP) followed by drug-primed reinstatement. The dose of d-AMP, 1mg/kg, was confirmed to significantly induce CPP. Using this dose, a group of rats was initially subjected to d-AMP CPP, which was followed by entry into an extinction protocol with an additional 3-day withdrawal before a drug-primed reinstatement test was carried out. Following extinction of d-AMP CPP, a lower dose of d-AMP, namely 0.75mg/kg, was able to significantly reinstate CPP. The BDNF mRNA levels in the selected brain areas were determined by real-time polymerase chain reaction (PCR) after the CPP and reinstatement. The BDNF mRNA level in the medial prefrontal cortex (mPFC) was significantly increased after the reinstatement, but not the CPP test. And, none of the other four assessed brain areas showed any change in BDNF mRNA level after d-AMP CPP or reinstatement. These findings support the notion that BDNF is involved in drug-seeking behavior and indicate that d-AMP reinstatement after extinction may be linked to an increase in BDNF mRNA expression in the mPFC.

Different effects of 5-HT receptor agonists on operant response in rats under DRL 10-s and DRL 30-s schedules.

5-hydroxytryptamine (5-HT ) is thought to be involved in a wide range of behavioral functions. Based on binding evidence, 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT), 1-(2,5-dimethoxy-4-indophenyl)-2-aminopropane (DOI), and m-chloro-phenyl-biguanide (m-CPBG) are selective 5-HT1a, 5-HT2, and 5-HT3 receptor agonists, respectively. The present study examined the effects of these 5-HT receptor agonists on operant behavior maintained on the differential reinforcement for low rate response 10-second (DRL 10-s) and 30-second (DRL 30-s) schedules of reinforcement. Water-deprived rats were trained to press a lever in response to DRL 10-s and DRL 30-s schedules. After a stable baseline was set, each subject was then repeatedly challenged using one drug with different doses through peripheral administration. The dose ranges were 0.025, 0.05, and 0.1 mg/kg (SC) for 8-OH-DPAT; 0.5, 1, and 2 mg/kg (SC) for DOI; and 1.3 and 9 mg/kg (IP) for m-CPBG. The overall results of the present work indicate that distinct profiles of operant response on DRL 10-s and DRL 30-s schedules were produced by 8-OH-DPAT, DOI, and, m-CPBG, based on quantitative and qualitative data analyses. All three 5-HT receptor agonists caused the number of responses to the DRL 10-s schedule to decrease significantly in a dose-related fashion. The operant performance on the DRL 10-s schedule was more sensitive to drug treatment than was that on the DRL 30-s schedule. Analyses of inter-response time (IRT) distributions revealed that different time bins were shifted by each of these three agents. The current data indicate that 8-OH-DPAT, DOI and m-CPBG can significantly alter operant response maintained on a DRL schedule. The distinct operant performance for each drug is believed to be derived from drug activation of its own specific 5-HT subtype receptors.

Effects of SCH23390 and raclopride on a run-climb-run behavioral task in rats.

The present study was designed to compare the putative differential behavioral consequences of treatment with SCH23390 (a selective dopamine D1 receptor blocker) and raclopride (a selective dopamine D2 receptor blocker) by employing a run-climb-run (RCR) behavioral task of different lengths. Rats were trained to traverse an uncovered floor alleyway (150 cm), climb a vertical rope (70 or 130 cm), and run across an upper board (100 cm) to access water for the reinforcement. At doses of 0.05, 0.10 and 0.15 mg/kg administered intraperitoneally 60 min before the behavioral session, both SCH23390 and raclopride significantly increased the total time to complete the tasks in a dose-related fashion. Microstructural analysis on the RCR behavioral performance revealed that the most apparent impairment induced by either drug was observed as the subject shifted motion from the end of the floor alleyway to the rope when hopping or to initiate climbing. However, the motion shift from climbing to running on the upper board was significantly impaired by raclopride, but not by SCH23390. Surprisingly, neither SCH23390 nor raclopride affected the climbing response itself. Running responses on the floor alleyway board were significantly disrupted by raclopride, whereas those on the upper board were significantly disrupted by SCH23390. Deficits induced by both drugs were more profound for the longer compared to the shorter rope, and were most notably shown at the transition area from running to climbing. These data indicate that both dopamine D1 and D2 receptors are involved in the RCR behavior performance. The results also suggest that the cost of motoric demand for behavioral performance is important for evaluating of the effects of drugs blocking dopamine receptors.

Operant performance following tail-pinch in the rat: effects of d-amphetamine.

To extend the investigation of tail-pinch induced behavioral changes, rats performing on a differential reinforcement of low rates of 10 sec (DRL10), a fixed-interval of 60 sec (F160), and a fixed-ratio of 20 (FR20) schedules were exposed to a paper clip applied to the tail. While a 10 min tail-pinch conducted 1 hr before operant sessions significantly altered the DRL10 behavior, this stressor had little effect on either F160 or FR20 responding. Marked DRL10 behavior performance changes following tail-pinch included increases in the number of lever presses, decreases in the number of the reinforcers, and disruption in the frequency distribution of inter-response times (IRT). These DRL10 operant deficits were diminished when the subject received a tail-pinch pretreatment followed by d-amphetamine treatment (0.2 and 2.0 mg/kg). In combination with biochemical data from others, the present results suggest that catecholamine systems are involved in modulation of DRL10 behavior following tail-pinch.

Distinct accumbal subareas are involved in place conditioning of amphetamine and cocaine.

In considering the heterogeneous function of the nucleus accumbens (NAC), the present work evaluated the conditioned place preference (CPP) after local infusion of d-amphetamine (AMP; 10, 15 microg/side) or cocaine (COC; 50, 100 microg/side) into two subareas of NAC, core and shell. A regular two-compartment CPP apparatus was used to test the place conditioning effects after 6 pairings of drug in one compartment and 6 pairings of vehicle in the other one. Significant CPP was observed with either AMP infused in the core area or COC infused into the shell area. Neither AMP in shell nor COC in core significantly produced CPP. These results indicate important differences between two neural substrates within NAC for the rewarding effects of AMP and COC on the CPP task.

Influences on water intake in the rat after lesions of the septal subareas.

It has been suggested that the septum plays an inhibitory role in the behavioral function. Recent work has shown that the septum is heterogeneous from the neuroanatomical perspective. The present study examined the water intake of rats lesioned with kainic acid (0.5 microg/0.5 microl/site) on three septal subregions: anterior medial (MSa), posterior medial (MSp), and lateral (LS) sites. Drinking volume was enhanced mostly in rats with the MSp lesion, and so was locomotor activity. However, these two measures were not significantly correlated. This polydipsia induced by MSp lesion was also found in a chronic domain. Another experiment further determined the dipsogenic effects of polyethylene glycol (PEG; 20%) and hypertonic saline (1 M NaCl) in MSp lesioned rats. Water intake increased significantly after administration of the hypertonic saline treatment, but not after injection of PEG. However, this disparity approached a nonsignificant level 8 hr after thirst challenges were conducted. In addition to revealing septal hyperdipsia. these data suggest that the septal subareas can be functionally heterogeneous in drinking behavior. The dipsogenic response profiles for the cellular and extracellular thirst challenges could be differentially affected by kainate lesion in the MSp.

Influence of SCH23390 and spiperone on the expression of conditioned place preference induced by d-amphetamine or cocaine in the rat.

The present study investigated the effects of selective dopamine D1 and D2 receptor antagonists, SCH23390 and spiperone, on the expression of conditioned place preference (CPP) induced by either d-amphetamine or cocaine. The CPP protocol consisted of three phases: pre-conditioning exploration, conditioning, and a post-conditioning test. The data indicated that CPP was significantly induced by intraperitoneal injection of either d-amphetamine (2 mg/kg) or cocaine (10 mg/kg). The expression of d-amphetamine CPP was significantly inhibited by SCH23390 (0.08, 0.16 mg/kg) and spiperone (0.15 mg/kg) when given alone before the post-conditioning test session. In contrast, such pretreatment to produce antagonistic effects was not observed for cocaine CPP. However, the expression of cocaine CPP was significantly attenuated by a combination of SCH23390 and spiperone administered prior to the test session. These data indicate that the rewarding properties of d-amphetamine and cocaine as expressed under the CPP task may depend upon different neural substrates. The degrees of D1 and D2 receptors involved in mediating the expression of CPP induced by d-amphetamine and cocaine are different.

Quantifying operant behavior deficits in rats with bilateral 6-hydroxydopamine lesions of the ventrolateral striatum.

The present study examined the effects of bilateral 6-hydroxydopamine lesions in the ventrolateral striatum on the operant behavior of rats. Use of the specially modified operant chambers allowed the measurement of forelimb response force and duration as well as the time intervals between selected behavior in the press-consume-press sequence. More specifically, four time intervals between separate behavioral events were measured: 1) the time from the end of forelimb response to entry of muzzle into the reinforcement well, 2) the time from muzzle entry to the first tongue lick of the water reinforcer; 3) the time from the last lick to muzzle withdrawal from the reinforcement well, and 4) the time from muzzle withdrawal to the beginning of the next forelimb operant response. As determined by neurochemical (HPLC) analysis, the lesioned group exhibited dopamine levels that were 35% of the control group. The operant behavioral deficits were most profoundly appeared in the first week of postoperative test. Behaviorally, the lesioned group exhibited longer forelimb response durations (bradykinesia), and decrements were seen in both the number of muzzle entries and the number of recorded licks during reward consumption. Furthermore, the lesion significantly increased the average latency to switch from the forelimb response to the entry of the muzzle into the reward well. The latency from well entry to the first tongue extension to the reward was also increased by the lesion. These data support the view that the rodent neostriatum is important in the control of behavioral sequences for psychomotor function and at the same time demonstrate the utility of new quantitative behavioral methods for investigating such functions.

Microinjections of SCH23390 and haloperidol into striatal subregions differentially affect water intake in the rat.

The nigrostriatal dopamine neurotransmission system is known to mediate various types of functional behaviors. Systemic administration of dopamine antagonists impairs drinking behavior induced by different dipsogenic agents. The present study was designed to further characterize the potential neural loci for hypodipsia under dopamine receptor blockade of the striatum via local administrations of SCH23390 and haloperidol. Water deprived rats were bilaterally implanted with cannula aimed at either the dorsomedial or ventrolateral striatum (DMS or VLS). Water intake was measured for 30 min after the intrastriatal microinjection. Results showed that the VLS site was more sensitive to DA receptor blockade than the DMS site to affect water consumption. However, only local infusion of haloperidol, rather than SCH23390, into the VLS site significantly reduced water intake. These data indicate that drinking performance can be sensitive to the heterogeneous functions of the striatum associated with specific dopamine receptor subtypes.

Chronic effects of haloperidol and SCH23390 on operant and licking behaviors in the rat.

Behavioral effects under chronic treatment of haloperidol and SCH23390 were examined for 18 days. Water deprived rats were trained either to perform an operant response on a fixed ratio 20 (FR20) schedule of reinforcement or to lick from a tube in a separate test. Both drugs completely impaired operant responding over all chronic administration days. Whether the lick performance was significantly affected differed depending upon the variable measured in that task. In contrast to the operant results, analysis of the microstructure of licking revealed very distinctive profiles of licking for each drug. Although the lick volume was consistantly reduced by both drugs across days, the decreased numbers of licks first observed were subsequently reversed back to the control level. However, the time courses and the reversal patterns were different for each drug. Haloperidol persistantly reduced the burst size for licking, whereas SCH23390 gradually enlarged it. For interlick interval (ILI) data, haloperidol had more impact on licks with longer ILI without influencing licks of shorter ILI. Contrarily, SCH23390 tended to lengthen the licks with shorter ILI without prolonging them enough to be classified as longer ILI licks. The dissociation of behavioral effects induced by chronic treatment of haloperidol and SCH23390 can be attributed to the drugs' blockade of different DA receptor subtypes. Also, the present study illustrates that using multiple behavioral tasks can be helpful to differentiate the subtle drug actions induced by DA receptor antagonists.

Distinguishing between haloperidol's and decamethonium's disruptive effects on operant behavior in rats: use of measurements that complement response rate.

The behavioral effects of haloperidol (0.04 to 0.16 mg/kg) and nonparalytic doses of decamethonium (0.2 to 0.8 mg/kg) were studied with operant methods that permitted the measurement of response rate, peak force of response, duration of response, and duration of the rat's head entry into the reinforcement dipper well. Type of operant response topography (forelimb press or forelimb grasp-and-pull) and peak force (low or high) required for reinforcement delivery were independent variables. The low-force, press-topography condition yielded qualitatively different profiles for the two drugs. Haloperidol increased peak force and duration of operant response, increased maximum head entry duration, and temporally dissociated forelimb and head entry behavior. Decamethonium decreased force and duration of operant response, did not appreciably affect maximum head entry duration, and did not influence the normal temporal coupling of forelimb and head entry responses. The haloperidol effects were seen as reflections of pseudo-Parkinsonism, not muscle weakness, which appeared to be the primary source of decamethonium's behavioral effects.

A new rodent model for neuroleptic-induced pseudo-parkinsonism: low doses of haloperidol increase forelimb tremor in the rat.

Rats were trained to use the right forelimb to exert continuous downward pressure on a force transducer and simultaneously to drink sweetened milk from a dipper controlled by the emitted force. Oscillations in forelimb force during this performance were spectrally analyzed to describe the tremorogenic effects of haloperidol (0.04, 0.08, or 0.16 mg/kg). Haloperidol reduced time-on-task and increased the variance of force oscillations in the 10.0-25.0-Hz frequency band. When atropine sulfate (5 mg/kg) was given along with haloperidol, time-on-task was partially restored, and the effects of haloperidol on the 10.0-25.0-Hz band were diminished. These data suggest that the behavioral deficits produced by relatively low doses of haloperidol in rats are analogous (and possibly homologous) to neuroleptic-induced Parkinsonian symptoms in humans.

Haloperidol produces within-session increments in operant response duration in rats.

On the basis of previously reported observations that haloperidol induces within-session decrements in operant response rate in rats, it was suspected that other measures of operant behavior may also display within-session changes after treatment with this neuroleptic. Accordingly, haloperidol (0.02, 0.04, 0.08, 0.16 mg/kg) was administered to six rats trained on a fixed-ratio 20 schedule of liquid food reinforcement, and response duration was recorded as a measure of drug effects independent of response rate. Significant within-session decrements in response rate and increments in response duration were observed as responding ensued. At the 0.08 mg/kg dose, 300% increases in response duration were seen during the last third of the responses made in a session. The progressive slowing of individual motor acts were interpreted as neuroleptic-induced Parkinsonism in the rat, and within-session decrements in response rate were likewise seen as a manifestation of the same pharmacological effects that increased response duration.

Effects of pimozide on emitted force, duration and rate of operant response maintained at low and high levels of required force.

The behavioral effects of pimozide (0.25, 0.5, and 1.0 mg/kg) were assessed in two separate experiments in which session mean peak force, maximum peak force, and response duration served as the dependent variables complementing operant response rate. In the first experiment, two groups of water-deprived rats were trained on a continuous reinforcement (CRF) schedule for reaching out and pressing downward on a force transducer with peak forces of at least 4 g (low-force group) or 40 g (high-force group). In the second experiment a pull-type response topography and fixed ratio 20 were used, and force requirements were 4 g for the low-force group and 100 g for the high-force condition. Under these conditions pimozide decreased response rate and increased response duration irrespective of response topography, required force, or schedule of reinforcement. Neither mean peak force nor maximum peak force were significantly decreased by the drug, and in the low-force CRF condition a small but significant dose-related rise in mean peak force was observed. It was hypothesized that neuroleptics exert their motor-impairing effects primarily in the temporal domain of behavior but do not appreciably affect the force dimension of performance capacity. And these temporal domain effects may be reflected in differences in the kinetic requirements for the overall behavior and not just the response itself. Additionally, the possibility that some of the observed effects could be accounted for by "anhedonia" was addressed.