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© Springer-Verlag 1997
O R I G I N A L I N V E S T I G AT I O N
&roles:F. Borsini · R. Cesana · J. Kelly · B. E. Leonard M. McNamara · J. Richards · L. Seiden
BIMT 17: a putative antidepressant with a fast onset of action?
&misc:Received: 10 March 1997 / Final version: 9 July 1997
&p.1:Abstract BIMT 17, the only compound reported to be a full 5-HT1A agonist and a 5-HT2A antagonist at the frontal cortex, was assessed in three animal paradigms sensitive to antidepressants in rats: olfactory bulbectomy (OB), differential-reinforcement-of-low rate 72-s (DRL 72-s) and learned helplessness (LH). In the OB rats, BIMT 17, given once daily for 14 consecutive days at an IP dose of 10 mg/kg, but not of 20 mg/kg, reduced the increase in ambulation of OB rats, 24 h after the last administration. In the DRL 72-s test, BIMT 17 had a different profile than imipramine. A single IP injection of 5, 10, 15 or 20 mg/kg BIMT 17, in contrast to the same doses of imipramine, did not affect response and reinforcement rate in DRL 72-s 1 h after the administration. On the other hand, BIMT 17 slightly shifted the peak of the interresponse time (IRT) distribution towards shorter IRT duration, while imipramine shifted the peak of the IRT distribution towards longer IRT duration. In the LH test, acute oral doses (36, 48 or 60 mg/kg) of BIMT 17, given 30 min before testing, reduced the number of escape failures in LH without altering the intertrial crossings. This effect was also induced by a repeated, but not single, administration with 8 or 16 mg/kg imipramine. The plasma levels following IP 10 or oral 48 mg/kg BIMT 17 were in the same range. These results indicate that BIMT 17 does not behave like imipramine in all the tests, and suggest that BIMT 17 acts through different mechanisms of action than imipramine. Only clinical trials will tell whether these mechanisms will be relevant, but if so, BIMT 17 might induce a faster onset of therapeutic activity.
&kwd:Key words BIMT 17 · Animal models of depression · Serotonin&bdy:
BIMT 17 (2H-benzimidazol-2-one, 1,3-dihydro-1-[2-[4[3-(trifluoromethyl)phenyl]-1-piperazinyl]ethyl]) (Fig. 1) has recently been described as the first compound capable of inhibiting forskolin-stimulated cAMP (Borsini et al. 1995b) and electrical activity (Borsini et al. 1995a) in the rat frontal cortex, by activating postsynaptic serotonin (5HT) 5-HT1A receptors and antagonizing 5-HT2A receptors (Borsini et al. 1995a,b). This mechanism of action in the frontal cortex has also been reported for antidepressants following chronic treatment (Borsini 1994). Thus, BIMT 17 may represent a prototype of a novel class of antidepressants with a potential faster onset of action. Interestingly, in the chronic mild stress model in mice, an animal model sensitive to antidepressants, a single administration of BIMT 17 had an effect similar to that following repeated treatment with fluoxetine (Willner 1995). An antidepressant-like effect of BIMT 17 has also been reported in the forced swimming test in mice (Cesana et al. 1995). The purpose of the present study was two-fold: a) to evaluate further the antidepressant potential of BIMT 17 in the rat and b) to confirm the faster onset of activity of BIMT 17. The effect of repeated treatment with BIMT 17 was evaluated in the olfactory bulbectomy (OB) model and the effect of a single administration of BIMT 17 in the
F. Borsini (u) · R. Cesana Boehringer Ingelheim Italia, Via Lorenzini 8, I-20139 Milano, Italy J. Kelly · B.E. Leonard · M. McNamara Pharmacology Department, University College Galway, Ireland J. Richards · L. Seiden Department of Pharmacological and Physiological Sciences, University of Chicago, 947 East 58th Street, Chicago, IL 60637, USA&/fn-block:
Fig. 1 Chemical structure of BIMT 17ig.c:&/f
2:This was according to Richards et al. Apparatus. BIMT 17 chloride was mixed with Tween 80 until a milky. The timing resolution of the system was 0. &p. positioned 90 cm above the floor of the apparatus. OB + 20 mg/kg BIMT 17. Specialities Septodont. and L. Drug administration. L. The doses of BIMT 17 refer to its salt form. The rats were housed two per cage in hanging stainless steel wire cages. the data was further analysed using the Student Newman Keuls test. calculated as a salt. This was administered IP at 1 ml/kg given a single daily dose between 0800 and 1000 hours of 10 or 20 mg/kg for 14 consecutive days. The animals were housed four per cage in plastic bottomed cages and were given a 1-week acclimatisation period prior to surgery. Animals. The open field consisted of a circular base. On non-training days (weekends).2:After surgery.2:Male Sprague-Dawley rats were obtained from Harlan Olac. These results are expressed as group mean and standard error of the mean.5 cm from the nearest side. in compliance with national and international laws and policies (EEC Council Directive 86/609. The operant chambers were 20. DRL 72-s This experiment was carried out by J. Lighting was controlled on a 12-h light: dark cycle (light period: 0800–2000 hours) and temperature was maintaned between 20 and 22°C.5 cm wide. After the operation. France). After completion of overnight DRL 72-s training the rats were trained during daily (5 days a week) 1-h sessions on the DRL 72-s schedule. Olfactory bulbectomy (OB). The operant chambers had grid floors. A lever was mounted 3 cm above the grid floor 4. FT 1-min schedule. Lights were on in the colony room from 0700 to 1900 hours. UK (weight on arrival: 230–250 g).2:Both BIMT 17 chloride and imipramine chloride (Ciba) were injected IP 1 h before the session in doses of 0. A solenoid operated dipper was located 10 cm to the left of the lever. 0600 and 0800 hours.1. At the time of surgery. &p. to evaluate further the possibility of a faster onset of action of BIMT 17.5 cm long. These results are expressed as group median and inter-quartile range. NIH Guide for the Care and Use of Laboratory Animals.. Reinforcement consisted of lifting the dipper (0. For sham animals. &p. The compounds Material and methods Procedures involving animals and their care were conducted in conformity with the institutional guidelines. Access to water was restricted to 20 min per day. = J L 358. J. Two drill holes of 2 mm diameter were made in the skull 5 mm rostral to the bregma and 2 mm lateral to the midline. S. &p. If any statistically significant change was found. &p. they were handled daily throughout the recovery period to eliminate any aggressiveness that would otherwise arise (Leonard and Tuite 1981). M. 10. and off when the training session ended. rats were individually marked with an ear notch for identification purposes. Training. rearing (number of times the rat simultaneously raised both forepaws off the floor of the apparatus). Sufficient pressure was applied to ensure that the periostium on the underlying bone had been penetrated. bleeding was controlled by plugging the holes with haemostatic sponge (Haemofibrine. the dura was carefully pierced and the wound closed. Illumination was provided by a 60-W bulb. Access to the dipper was through a round 4. The rats were then shifted to a DRL-72-s training regimen. DRL 72-s overnight training consisted of six 1-h sessions with a 30-min time-out (house light off) between each session. Rats which did not acquire the lever press response after five overnight training sessions were manually trained to press it.15 N was required for a lever press to be detected. Oxytetracycline dusting powder was sprinkled on the wound prior to closure. All measurements were carried out in a darkened room on the morning of day 15 between . (1977). Each animal was placed in the centre of the open field apparatus and the following parameters were measured over a 3-min period: ambulation (number of squares crossed). and B. The animals were allowed to recover for 14 days following surgery. 12. Body weights of each animal in all the groups were measured at days 1 and 15. Wisc. Dec. Bulbectomized rats This experiment was carried out by M. sham + 20 mg/kg BIMT 17. (1994). the “learned helplessness” (LH) model was used. The operant chambers were enclosed in 80 quart Coleman ice chests to attenuate external stimuli. “Open field”.2:Fifteen male Sprague-Dawley rats (Hotzman. 15 and 20 mg/kg. USA) weighing between 350 and 500 g at the time of drug administration were used. K. OB + 10 mg/kg BIMT 17. The rats had received daily 1-h DRL 72-s schedule sessions for 10 weeks before drug testing was initiated. 5. to give a concentration of either 10 or 20 mg/ml. Drug administration. OB + vehicle.379 differential-reinforcement-of-low rate 72-s (DRL 72-s) paradigm.5% w/v 2-2–2 tribromoethanol (10 ml/kg IP) essentially as described by Cairncross at al. 20. Animals. The head was shaved and a midline sagittal incision was made extending at least 1 cm rostral to the bregma. the olfactory bulbs were aspirated using a water suction pump. Kruskal-Wallis test was performed on the data relative to the “open field” test.2:The rats were tested on day 15 of experiment.5 cm deep and 23. 85–23. The stimulus light was turned on when a training session began.5 cm diameter hole in the front panel. the rats were given 20 min access to water between 1000 and 1400 hours.2:Two-way analysis of variance was performed to evaluate body weights. grooming (number of times the rat stopped and groomed itself) and defaecation (the number of faecal boli deposited). the rats were allocated to six groups (nine animals in each group): sham + vehicle. Madison. E. Saline was then added. 90 cm in diameter which was divided into 10 cm squares by faint yellow lines. The rats were trained overnight on the DRL 72-s schedule for 10 nights.2:Upon arrival in the colony the rats were adapted to the 20 min per day access to water regimen for 1 week. sham + 10 mg/kg BIMT 17. 1987. The schedule contingencies were programmed using the SKED-11 software system. &p. &p.025 ml) from a water trough to within reach of the rat’s tongue for a period of 4 s. these data were then tested using a MannWhitney U-test. For OB animals. 1985). A downward force of approximately 0.01 s. Care was taken not to damage the frontal cortex. &p. homogenous dispersion was obtained. On training days the rats received 20 min access to water at the end of their training session. This apparatus is essentially as described by Gray and Lalljee (1974). Food (4% Tek Lab rat chow) was available ad lib. They were allowed free access to food and water throughout the course of the experiment. &p. If any statistically significant changes were found. A stimulus light mounted 15 cm above the floor on the back wall of the chamber provided the only illumination within the chamber. Controls received injections of vehicle alone. The wall surrounding the base consisted of a 75 cm high aluminium sheet. In addition. The operant chambers were connected to a PDP-11/73 micro computer via a Coulbourn Lablinc interface. Fans mounted on the ice chests provided ventilation and masking noise.. The rats were then trained to bar press in over-night training sessions using an alternative FR1. aluminium front and back walls and Plexiglas sides. NIH publication no.2:Bilateral olfactory bulbectomy was performed in rats anaesthetised with 2. and the effect of a single administration of BIMT 17 was evaluated in comparison to single and repeated treatments with imipramine. Data analysis. R.
.2:The shuttle-box test was carried out according to Geoffroy et al. &p. Data analysis. randomized inescapable shocks (15 s duration. IS+ 24. The animals were kept in the experimental room with a regulated environment (21 ± 1°C. On day 4. 4. The overall level of significance was set at P<0. Control values (average of the Thursdays which occurred during each drug’s dose response determination) for each measure were compared to each drug dose (including vehicle) using within group t-tests. Imipramine was given either acutely or repeatedly. The effect of treatment versus IS+ group on the number of escape failures was analysed by the Kruskal-Wallis test followed by the Dunn test. Cary. &p. Ugo Basile). IS+ 0. In the first ten trials the rats were required to change side once (FR1) and in the following 15 trials the rats had to change side twice (FR2) in order to terminate both shock and light. &p. Decreases in the value of PkA indicate that the rats are responding in a less coherent fashion and that the distribution of waiting times (IRTs) is more dispersed. Water was then incrementally added to obtain the appropriate drug dose. The bars to the right of the burst component indicate the pause component of the IRT distribution (IRTs ≥ 6s). Four two-way shuttle-boxes (Ugo Basile.2:Naive male Crl:(WI)BR Wistar rats (Charles River Italia S. The statistical evaluations were carried out with the program system SAS (SAS Institute Inc. C. Control performance was the average of the Thursdays which occurred during each drug’s dose-response determination. Animals. Rats well trained on the DRL 72-s schedule generate IRT distributions which have well defined peaks. The Wilcoxon two-sample test was used to compare the results relative to the control groups (IS+ versus IS–). Italy) weighing 200–250 g were used. every 30 ± 24 s) to the grid floor.05. and peak location (PkL) measures were determined for each rat..07 on a DEC Computer.8-mA scrambled electric shock). 2 and 3 and the test compound in the last injection (day 4). The test took place on day 4 between 08. in fact. The peak location (PkL) is the median IRT duration which bisects the shaded region above the corresponding negative exponential. that rats exposed to inescapable shock often do not fail to learn FR1 shuttle-box response. Control groups (IS– and IS+) received the corresponding vehicle treatment. Control rats were placed in identical chambers for 45 min but shock was not given (IS–). Version 6. mod. The experimental groups were 6 (IS–. Doses of imipramine refer to the base. Eight of the rats received the BIMT 17 dose determination first. (1990). 50–55% humidity and 12-h light-dark cycle. followed by the imipramine dose determination. The remaining seven rats received the imipramine dose determination first. The repeated treatment with imipramine was performed as follows: the first injection was done 2 h after the session of inescapable shock (day 1). the animals were injected 30 min before the shuttle-box test.2:Three different experiments were conducted: one with BIMT 17 base given acutely and the others with imipramine given acutely or chronically.). After determining that there was no effect of treatment order. All the helplessness induction trials were performed on day 1 in the morning. North Carolina).p. injections were given twice daily (0800 and 1600 hours) on day 2 and 3. The vehicle for imipramine was saline. PkL indicates the central location of the peak.group on the number of intertrial crossings was analysed by the Kruskal-Wallis test. Imipramine hydrochloride (Sigma) was dissolved in saline and given IP at doses of 4.30 and 12. light on at 7. The connected dots indicate the expected appearance of the pause component of the IRT distribution if the rat emitted the same number of responses. A two-factor ANOVA. &p.. The compound was solubilized in the following vehicle: 25% v/v polyethylene glycol-400 acidified (1. After an initial 5-min habituation. The shock duration was 15 s max with an intertrial time of 25 s. in a volume of 5 ml/kg. In the experiment with BIMT 17. was done for both BIMT 17 and imipramine to determine if treatment order affected the results.2:Response rate. BIMT 17 and imipramine were tested using a cross-over procedure. 7502) were used. 1. 8 and 16 mg/kg on a volume of 2 ml/kg. Richards et al. Data analysis. with treatment order as a betweengroups factor and drug dose as a within-groups factor. VMS.2:The behavioural data (FR2 escape failures and intertrial crossings) were expressed as medians and interquartile range. PkA indicates the area of the IRT distribution peak and PkL indicates the location of the IRT distribution peak. The animals were housed two per cage in makrolon cages (42×26×15 h cm) with free access to water and food (4RF21 pellets “Mucedola Srl”. peak area (PkA). An increase in PkL indicates the rats are waiting longer. The number of escape failures (i. It has already been noted. Tween 80. In Fig. the data were combined and analyzed using within group t-tests. &p. reinforcement rate. IS+ 36. the failure to respond during the 15-s shock on) and the number of intertrial crossings (i. Acute treated rats received vehicle on day 1. The PkA and PkL metrics quantitatively characterize the profile of the interresponse time (IRT) distributions generated by responding on the DRL 72-s schedule (Richards and Seiden 1991.380 were dissolved in vehicle (V) to form a solution of 1 ml/kg for injection. Two experimental blocks per week were carried out. The doses of BIMT 17 and imipramine were given in ascending order. All four dependent variables (response rate.e. This expected curve is called the corresponding negative exponential. 80 mg/ml water. whereas the FR2 response is always reduced (Maier and Seligman 1976). The effect of treatment versus IS. o. PkA is indicated by the shaded region of the obtained IRT histogram above the corresponding negative exponential. followed by the BIMT 17 dose determination.A. Settimo Milanese. 96 rats were divided into four experimental blocks of 24 rats each (four rats for each group). peak area and peak location) were examined in this fashion. BIMT 17 was combined with Tween 80 in a ratio of 1 mg BIMT 17 to 4 mg Tween 80. but randomly in time with respect to the preceding response.s. 1993a). For the evaluation of rat’s performances. The single shaded histogram bar on the left indicates the burst component of the IRT distribution (IRT < 6 s). The aim of the latter comparison was to verify if the motility of the drug-treated animals was greater than the motility of unshocked rats. A glass vial and a small magnetic stir rod were used to obtain a homogeneous dispersion.e. a total of 25 unsignalled escape trials were presented to each animal (each trial started with the simultaneous onset of a light and a 0. Drug doses were prepared immediately before administration. Learned helplessness This experiment was carried out by R. A decrease in the value of PkL indicates the rats are not waiting as long between responses.00 a. Electric footshock was delivered to the animals by mean of eight chambers (22×22×27 h cm) with grey Plexiglas walls and covers and with the floor consisting of stainless-steel grids (Ugo Basile). &p. This result indicated that there was no effect of treatment order. Helplessness (LH) induction. Calco. the number of side-to-side crossing during the 25-s shock off) was recorded by mean of a computerized data acquisition system (Basilink. twotailed.m.30 a.. Drug administration. reinforcement rate. only the FR2 escape responses were analysed. the BIMT 17 vehicle histogram (top left histogram) illustrates how the profile of the IRT distributions was quantitatively characterized by PkA and PkL. was used for the BIMT 17 vehicle injection. IS+ 48 and IS+ 60 mg/kg BIMT 17 base) of 16 rats each. .m.2 mA. This procedure resulted in a clear fluid with no visible particles. Drugs were administered on Tuesday and Friday of each week. Shuttle-box test. Increases in the value of PkA indicate that the rats are responding in a more coherent fashion and that the IRT distribution is less dispersed. 30 min before testing. Italy). The results of this analysis showed no significant between group differences. BIMT 17 was given orally. A Bonferroni correction was used to guard against a type 1 error due to multiple comparisons.2:Rats were randomly assigned to groups and subjected to one session of inescapable footshock (IS+).5% citric) distilled water. A constant-current shocker delivered 60 scrambled.
03.56. 5).05 versus Sham-vehicle.14.5 ml water. Thus. 48) = 0.272]. Learned helplessness . 70) = 0. conditioned with 2.0001] toward longer IRT durations in a dose-dependent fashion (Fig. The imipramine induced shift of PkL toward longer waiting times is graphically shown in the IRT distribution presented on the right side of Fig. The dried samples were taken up in mobile phase and injected into the HPLC system (Perkin-Elmer).0001] and increased [F(5. Chronic 10 mg/kg BIMT 17 (P<0. 70) = 1. 48) = 0. # P<0.7. 48) = 1.26 ml/min. ISS 200 autosampler. DRL 72-s There was no significant effect of acute BIMT 17 treatment on response [F(5. Three different groups of animals (six rats/group) were administered IP with 10 or 20 mg/kg or orally with 48 mg/kg. BIMT 17 shifted the peak of the IRT distribution [F(5. 24 h after the last injection. P < 0. 2).5 ml methanol followed by 2.381 Plasma levels of BIMT 17 Plasma samples (0. Rats were placed in the “open field” after 2 weeks of treatment with BIMT 17 (IP. of nine animals. BIMT 17 and internal standard were separated with a reverse phase ODS Hypersyl column (100×2. 4. The analytes were eluted twice with 1 ml ammonia solution in methanol (33%) and taken to dryness under nitrogen stream at about 45°C.34. P = 0.. 70) = 3. and interquartile range (in brackets). the lesion effect was still evident [F(1. Detection was UV at λ = 210 nm. grooming or defecation (data not shown). 48) = 5. P < 0.000]. 70) = 7. was used but produced an almost catatonic state and failures in nine of 15 rats to respond in the operant chamber 1 h after administration. In contrast. P < 0. 4) shows that the BIMT 17 and imipramine had very different effects on the IRT distribution profiles. The BIMT 17-induced shift of PkL toward shorter waiting times is graphically shown in the IRT distributions presented in Fig. 70) = 1. or lesion*drug interaction [F(2. imipramine had no effect on PkA [F(5.5 M KH2PO4. or lesion*drug interaction [F(2. but no drug effect [F(2. Examination of the histograms (Fig. but there was no drug effect [F(2. 3). P = 0. vortexed and added to solid-phase extraction cartridges. In contrast. The reversal of the escape deficit was obtained without affecting the number of intertrial crossings (Table 1). The single adFig. respectively (P<0. BIMT 17 decreased PkA [F(5.64.0001] at 15 and 20 mg/kg: such a decrease is reflected in the IRT histograms presented on the left side of Fig. The effect was statistically significant after administration of 48 mg/kg BIMT 17. adjusted to 3 ml with 0. 3. 4). This effect was significant at the 15 mg/kg dose.37. grooming or defecation in sham or OB rats (data not shown). P = 0. once daily). 3). in contrast.05) (Fig. Column temperature was set at 40°C and flow rate was 0. whereas imipramine decreased [F(5. Mobile phase consisted of 28% (v/v) of acetonitrile and 72% of the following acqueous solution: 10 mM KH2PO4 and triethylamine adjusted at pH 3 with 2 M H3PO4. At this time. i. 3).18] and reinforcement [F(5. consisting of a Series LC 250 pump. A typical hyperactivity was observed in OB rats when placed in the “open field” apparatus when compared to shams (P < 0. A higher dose of BIMT 17. approximate retention time for BIMT 17 and internal standard were 10 and 13 min. Fig. The cartridges were washed twice with 1 ml of a mixture acetonitrile/water (1:1. 70) = 27. P < 0.5 ml) were addedd with 100 µl of internal standard (1 µg/ml BIMT 31).970]. P = 0. 48) = 22.1 mm i. Results are expressed as median. imipramine shifted the peak of the IRT distribution [F(5. On day 15. the mean body weights (g ± SEM) of the control OB group were reduced when compared to its sham group prior to treatment (344 ± 6 and 370 ± 9.873]. HewlettPackard). 70) = 12.98.e.c:&/f Results Bulbectomized rats Prior to drug treatment. P = 0. P = 0.0001] these parameters. Plasma levels were collected 60 min after the IP injections or 30 min after the oral administration. 40 mg/kg. under these chromatographic conditions. 70) = 15. respectively. v/v) and dried under vacuum for about 30 min. respectively (top panel on The acute oral administration of BIMT 17 caused a reduction (Kruskal-Wallis test P = 0.05 versus OB-vehicleig.695]. BIMT 17 increased the dispersion of the waiting time distribution (Fig. 3). 2).01). as demonstrated by a significant lesion effect [F(1. * P<0. no significant differences were found in rearing.62] (Fig. The test was carried out on day 15.04. UV/Vis detector mod LC 95 and Turbochrom data collection system. respectively).d.65. 4. 48) = 0. 2 Effect of BIMT 17 in the “open-field” in sham and OB rats. P < 0. without affecting rearing.61] rate up to a dose of 20 mg/kg (top panel on Fig.01] toward shorter IRT durations (bottom right panel of Fig.041) in the number of escape failures of helpless rats during the shuttle-box test session (Fig.05) but not 20 mg/kg BIMT 17 treatment significantly reduced the increase in ambulation of OB rats (Fig.018]. 3).14. the mean body weights for OB and sham control groups were 361 ± 13 and 417 ± 9.
c: Group Acute bimt 17 IS− IS+ IS+ IS+ IS+ IS+ Acute imipramine ISIS+ IS+ IS+ IS+ Repeated imipramine IS− IS+ IS+ IS+ IS+ Treatment Dose mg/kg No.5–4) (0–0. 30 min before testing on day 4.27 ± 0.5 1.05 &/tbl.36. Plasma levels The plasma levels (µg/ml) of BIMT 17 60 min after its IP administration with 10 or 20 mg/kg were (mean ± SEM) 1. 4).0 0. and peak location.5) (1–4.5 0. 3 Effects of BIMT 17 and imipramine on response rate. Peak area and location are metrics designed to measure the IRT distribution profile (see text and Fig.59 ± 0. peak area. Imipramine was given as a single dose 30 min before testing. Repeated administered imipramine was given i. and repeated imipramine on the intertrial crossings in the LH test in rats&/tbl. 8 and 16 mg/kg imipramine (Fig. of intertrial crossings median (interquartile range) BIMT 17 was given orally 30 min before testing.0 2.5) (0–1. imipramine and vehicle were given IP 1 h before the session.5 0 0.5) (0. 2 h after inescapable shock on day 1.05 and 1. 8 and 16 mg/kg being the active doses.5) (0–0.0 2.382 Fig. twice daily on days 2 and 3. Group sizes were constituted by 16 rats for BIMT 17 experiment and by 8 rats for the imipramine experiments. 8 and 16 mg/kg imipramine decreased the number of escape failures of helpless rats during the shuttle-box test session (Kruskal-Wallis test P = 0. whereas 30 min after its oral administration with 48 mg/kg they were (mean ± SEM) 1.88).05 versus respective V-groupig.5 0 0 0 0.18.p. The repeated administration of 4.5 2.5–2) (0–1) (0–0) (0–0) (0–1) (0–2. KruskalWallis test: P >0. * P < 0. 5) was completely devoid of effects (Kruskal-Wallis test P = 0.5 (0–1) (0–3) (0–1) (1–4. reinforcement rate. BIMT 17.c:&/f Table 1 Effect of acute BIMT 17 and imipramine.0 0 0.5) (0–1) ministration of 4. Imipramine was given as a single or a repeated administration.5 2. respectively.001).5) (0. without affecting the number of intertrial crossings (Table 1).: Vehicle Vehicle BIMT 17 BIMT 17 BIMT 17 BIMT 17 Vehicle Vehicle Imipramine Imipramine Imipramine Vehicle Vehicle Imipramine Imipramine Imipramine 0 0 24 36 48 60 0 0 4 8 16 0 0 4 8 16 0.24 ± 0. .
Both the OB and LH models only permit the observation of very strong effects. Repeated administration of BIMT 17 at a dose of 10 mg/kg significantly reversed the increased ambulation associated with the OB rat in the “open field” test. and this finding has been confirmed in the present study where IP 10 mg/kg and oral 48 mg/kg gave similar plasma levels. Interestingly. Similarly. The dashed vertical line indicates the 72-s IRT duration requirement for reinforcement. 1995). Reversal of increase in ambulation in OB rats has also been reported for antidepressants. Song and Leonard 1994). as also evidenced by DRL 72-s. OB rats have an altered 5-HT metabolism in the frontal cortex (Lumia et al. Bulbectomized rats The ablation of the olfactory bulbs induces many behavioural changes in the rat (Leonard 1984. the single histogram bar at the far right indicates the relative frequency of IRTs > 144 s in the tail of the obtained IRT distribution. both typical and atypical including selective serotonin reuptake inhibitors. but only the lowest dose induced a statistically significant effect. Van Riezen and Leonard 1990). Alternatively. 4 Interresponse time (IRT) histograms visualizing how BIMT 17 and imipramine effected the DRL 72-s IRT distribution profiles. The single dot at the far right of the histogram indicates the relative frequency of IRTs > 144 s predicted to occur in the tail of the corresponding negative exponential. shaded histogram bar on the left) indicating bursting and a second mode at longer IRT durations (IRT ≥ 6 s. This might explain why the effect of BIMT 17 in the OB model apparently appears similar at both doses. Jesberger and Richardson 1986). 1977. where the highest dose exerted an effect which was smaller than that observed at a lower dose. and in oth- . BIMT 17 is less potent than when given IP (Cesana et al. Leonard 1984. The histograms represent the averaged relative frequencies of 15 ratsig. The IRT distributions are bimodal with one mode at short IRT durations (IRT < 6 s. The BIMT 17 vehicle histogram (top left histogram) illustrates how the IRT distribution profile is quantitatively characterized by the peak area (PkA) and peak location (PkL) metrics (see text for explanation). open histogram bars) indicating pausing. The primary index of potential antidepressant activity is reversal of increased ambulation scores (Jancsar and Leonard 1983. the data were expressed as median and interquartiles because the distribution of values was not Gaussian. but not for centrally acting drugs which lack antidepressant activity (Jancsar and Leonard 1983. The triangle in the burst category indicates the relative frequency of burst responses predicted by extrapolation of the corresponding negative exponential into the burst component. which produces similar plasma levels of 10 mg/kg IP) interfere with rat performance. it may be that doses greater than 10 mg/kg IP (or 48 mg/kg PO. 10 and 20 mg/kg. due to either the small window for drugs to restore a normal behaviour or to the variability of data. 1984.383 Fig. Leonard 1982. 1992).c:&/f Discussion Given orally. In fact. Similar consideration holds with the LH model. Song and Leonard 1994): repeated treatment with these antidepressants seems to be necessary to observe an effect (Van Riezen et al. This second mode shows that the rats have learned to wait between responses (although not quite long enough). in both tests.
In fact. but not after acute. 1995a). an agonist at 5-HT1A receptors and an antagonist upon 5-HT2A receptors in the frontal cortex (Borsini et al. # P < 0. reversal of LH is obtained after repeated. BIMT 17 shifted the PkL toward shorter IRT durations while imipramine shifted PkL toward longer IRT durations.b). Marek et al. amphetamine) but in contrast to antidepressants. BIMT 17 induced an antidepressant-like behavioural effect in the LH paradigm after a single administration whilst antidepressants must be given repeatedly. 1995a. a 5-HT2A antagonist and 5HT1A agonist in the cortex. showed an antidepressant-like effect in the OB model following chronic administration. Learned helplessness Among the animal models of depression. 1985. DRL 72-s The profiles of the acute effects of BIMT 17 and the antidepressant imipramine on DRL 72-s performance are very different. 1993b) has shown that a variety of antidepressant compounds shift the peak of the IRT distribution toward longer IRT durations without decreasing PkA. and interquartile range (in brackets). imipramine (Ceci et al. 1993) increase the firing rate of frontocortical neurons.b) differentiates it from the other agonists. BIMT 17 has a direct action on postsynaptic 5-HT receptors (Borsini et al. 1993) and. BIMT 17 had no effect on response and reinforcement rate while imipramine decreased response rate and increased reinforcement rate. 1995a. 1995a. The OB rodent model has been suggested as a model for agitated hyposerotonergic depression (Lumia et al. 5 Effect of BIMT 17 and imipramine on the escape failures in the LH test. Single doses of 8-OH-DPAT and buspirone (Borsini et al. but not imipramine. It has been suggested that antidepressants may act in DRL 72-s by blocking 5-HT2 receptors and activating 5HT1A receptors (Marek and Seiden 1988b. BIMT 17. This shift in the IRT distribution toward longer waiting times results in an increased reinforcement rate. whereas BIMT 17 decreases the firing rate by activating 5-HT1A receptors (Borsini et al. Previous re- search (Seiden et al. BIMT 17. selective 5-HT reuptake inhibitors and electroconvulsive shock. 30 min before testing on day 4. The fact that BIMT 17 is the first full postsynaptic agonist (Borsini et al. in these animals. Although BIMT 17 did not affect response and reinforcement rate at the doses tested. Reversal of LH is also obtained after acute administration of psychostimulant compounds (e. 1992) or fluoxetine (Ceci et al. induced decreases in PkA. BIMT 17 (top panel) was given orally 30 min before testing. It may be that this difference in the mechanism of action can explain the different effect of BIMT 17 in comparison with antidepressants in this animal model. 1992). repeated administration with amitriptyline and trazodone had the common effect of reducing the number of 5-HT1A receptors in the frontal cortex but not in other areas (Gurevich et al. By these criteria. Imipramine was given as a single ( middle panel) or a repeated (bottom panel) administration. In addition. 1993). 1989a. . The shift in the peak toward shorter waiting times and the increased dispersion of the waiting times indicates that BIMT 17 disrupted performance at doses greater than 10 mg/kg. Even if not all 5-HT1A agonists exert an antidepressant-like effect in this test (Richards et al. whereas all the other compounds referred to seem to activate presynaptic 5-HT mechanisms. OB mice showed an increase in 5-HT2 receptors only in the frontal cortex (Gurevich et al. * P < 0. Repeated administered imipramine was given IP 2 h after inescapable shock on day 1. In addition. the LH possesses an interesting degree of “face validity” and “predictive validity” (Willner 1991). Richard et al.b). twice daily on days 2 and 3. Results are expressed as median. 1995). Thus. monoamine oxidase inhibitors.01 versus IS+ (Wilcoxon two-sample test). it should be pointed out that 8-OH-DPAT and buspirone do not behave as agonists in the frontal cortex (Borsini et al. administration of tricyclic antidepressants.384 Fig. 1994).01 versus vehicle-treated group (Dunn test)ig. Group sizes were constituted by 16 rats for BIMT 17 experiment and by eight rats for the imipramine experiments. Marek and Seiden 1988a. In fact. increasing the number of intertrial crossings (Christensen 1993). BIMT 17 does not resemble the acute effects of antidepressants on the DRL 72-s screen. did not affect DRL 72-s behaviour. The reversal of LH observed after the single administration of BIMT 17 did not seem to depend on modification of the number of intertrial crossings. it is surprising that BIMT 17. Cesana et al. 1995b). 1995a). Imipramine was given as a single dose 30 min before testing.g. it did significantly affect the IRT distribution profile.c:&/f er brain regions. they also affect locomotor activity. Thus.
General discussion The purpose of this study was two-fold. Thus. Baschirotto A. Firstly. by using the LH paradigm where it was found that BIMT 17 induced antidepressant-like effect after a single administration. imipramine was also active in the LH. de Montis MG (1995) Imipramine and fluoxetine prevent the stress-induced . These differences are not surprising. Donetti A (1995b) BIMT 17. the forced swimming test (Cesana et al. Synapse 15: 243–245 Barone P. directly activates postsynaptic 5HT inhibitory responses in the cortex. Martin P. It is worth noting that BIMT 17 and not imipramine induced an effect in the LH. 1995). Borsini F (1993) Effect of fluoxetine on the spontaneous electrical activity of fronto-cortical neurons. these results indicate that BIMT 17 does not behave like imipramine in all the tests and suggest that BIMT 17 acts through different mechanisms of action than imipramine. 1995). a 5-HT 2A antagonist and 5HT1A receptor full agonist. Eur J Pharmacol 183: 1900–1901 Borsini F (1994) Balance between cortical 5-HT 1A and 5-HT2 receptor function: hypothesis for a faster antidepressant action. when the postsynaptic component of imipramine is unmasked after repeated treatment. are more effective on the dendro-somatic region than on the postsynaptic 5-HT receptors (Adell and Artigas 1991. whereas imipramine and not BIMT 17 induced an effect on DRL 72-s. in rat cerebral cortex. &p. taken as a whole. Taddei A. In conclusion. Naunyn-Schmiedeberg’s Arch Pharmacol 352: 283–290 Borsini F. Donetti A (1995a) BIMT 17 a 5-HT 1A agonist/5-HT2A antagonist. BIMT 17 was tested in OB rats. such effects correlate with antidepressant concentrations in the anterior neocortex (Petty et al. the different activities of acute BIMT 17 and imipramine may depend on their postsynaptic (BIMT 17) and presynaptic (imipramine) component. but if so. to confirm the faster onset of action of BIMT 17. the different effect of BIMT 17 and the 5-HT uptake inhibitors may depend on this different location of action. Nakai et al.385 It is worth noting that a supersensitivity of 5-HT2 receptors possibly related to LH behaviour has been suggested and that the repeated administration of antidepressants induces a down-regulation of these receptors which parallels the reversal of LH behaviour (Barone et al. NaunynSchmiedeberg’s Arch Pharmacol 352: 276–282 Cairncross KD. it has been reported that fluoxetine. In addition. Artigas F (1991) Differential effects of clomipramine given locally or systemically on extracellular 5-hydroxytryptamine in raphe nuclei and frontal cortex. Tagliamonte. Moreover. 1982). as has already been seen in the chronic mild stress procedure (Willner 1995). in the LH procedure and in the DRL 72-s model and induced antidepressant-like effects in the former two paradigms but not in the latter one. 8OH-DPAT. Atger F. Bel and Artigas 1993). Baschirotto A. 1994) and that tricyclic antidepressants normalize this change (Sherman and Petty 1982). Thus. References Adell A. The prefrontal cortex was the only brain region in which microinjection of 5-HT reversed learned helplessness 1 h after injection (Petty and Sherman 1980). Ghieri O. Artigas F (1993) Chronic treatment with fluvoxamine increases extracellular serotonin in frontal cortex but not in raphe nuclei. Antonini G. Interestingly. Only clinical trials will tell whether these mechanisms will be relevant. Artigas 1993. 1990. after repeated treatment. Borsini F (1992) Effect of 5HT monoamine uptake blockers and 5HT1A agonists on spontaneous firing rate of prefrontal cortical neurons. Giraldo E. Fillion G (1990) Increased number of 5HT2 receptors in the “learned helplessness” rat. produces an effect when microinjected at postsynaptic receptors in the LH paradigm whereas the action of a low dose of fluoxetine in the DRL 72-s paradigm is reduced after destruction of 5-HT-containing neurons by a selective 5-HT neurotoxin (Marek et al. after a single administration. BIMT 17 might induce a faster onset of therapeutic activity. This may be also substantiated by the fact that the 5-HT1A agonist. Secondly. A study to investigate the exact mechanism of action of BIMT 17 in the LH is currently under investigation. it has been reported that LH induction modifies in vivo cortical 5HT release (Petty and Sherman 1983. an effect common to antidepressants (see Borsini 1994). Puech A. 1988) and it has been shown that their effect is due to the activation of postsynaptic 5-HT1A receptors (Martin et al.2:Acknowledgements This study was mostly supported by IMI contract no. to confirm the potential antidepressant activity of BIMT 17 as previously shown in two animal paradigms sensitive to antidepressants. 1990). 1989a). Monferini E. 1995) and the chronic mild stress procedure (Willner 1995). it is not surprising that BIMT 17 can reverse the behavioural deficit of rats in the LH after a single administration.1 Ceci A. Cox B. a new potential antidepressant. Bietti G. Parenti M. Bietti G. Wren AF. 5HT uptake inhibitors. Physiol Behav 19: 485–487 Ceci A. Behav Pharmacol 6: 688–694 Christensen AV (1993) Learned helplessness: an animal model of depression? Neuropsychopharmacology 9: 167S Gambarana C. Similar results with fuoxetine and the 5-HT neurotoxin were also obtained in the LH test (Martin et al. 1991). although a different experimental protocol of the test was used (Gambarana et al.e. as these two compounds may exert different effects on the 5-HT-containing neurons. Thus. Naunyn-Schmiedeberg’s Arch Pharmacol 343: 237–244 Artigas F (1993) 5-HT and antidepressants: new views from microdialysis studies. Pharmacol Res 30: 1–11 Borsini F. Ceci A. Trends Pharmacol Sci 14: 262 Bel N. Schnieden H (1977) Effects of olfactory bulbectomy and domicile on stress induced corticosterone release in the rat. 1990. the helpless behavior is reversed in the LH model by repeated treatment with putative 5-HT1A agonists (Giral et al. Ciprandi C. i. in the forced swimming test in mice. Petty et al. Due to the serotonergic activity of BIMT 17. 55312/46. Soc Neurosci Abstr 18: 317. Interestingly. Eur J Pharmacol 250: 461–464 Cesana R. Borsini F (1995) The effect of BIMT 17. prevents the escape deficit in this animal model also by activating 5HT1A receptors.
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