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The Role of 5-HT2A and 5-HT2C Receptors on Harmaline- Induced

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The Role of 5-HT2A and 5-HT2C Receptors on Harmaline- Induced Iranian Journal of Veterinary Research, Shiraz University, 2013, Vol. 14, No. 2, Pages 94-99 The role of 5-HT 2A and 5-HT 2C receptors on harmaline- induced eating behavior in 24-h food-deprived broiler cockerels Zendehdel, M. 1*; Mokhtarpouriani, K. 1; Babapour, V. 1; Pourrahimi, M. 2 and Hamidi, F. 1 1Department of Basic Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran; 2MSc in Physiology, Department of Biology, Faculty of Basic Sciences, University of Damghan, Damghan, Iran *Correspondence: M. Zendehdel, Department of Basic Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran. E-mail: [email protected] (Received 4 Oct 2011; revised version 9 Jan 2013; accepted 14 Jan 2013) Summary This study was designed to examine the effects of intracerebroventricular (ICV) injection of ketanserin (5-HT 2a receptor antagonist) and SB242084 (5-HT 2c receptor antagonist) on harmaline induced feeding and drinking response in 24-h food-deprived (FD24) broiler cockerels. At first, guide cannula was surgically implanted in the right lateral ventricle of chickens. In experiment 1, birds were injected intracerebroventriculary with 0, 25, 50 and 100 µg of harmaline. In experiment 2, chickens received 10 µg ketanserin prior to the injection of harmaline. In experiment 3, birds were administered with harmaline after 3 µg SB242084 and the cumulative food and water intake was determined at 3 h post injection. The results of this study showed that harmaline decreases food consumption and increases water intake in FD24 broiler cockerels (P ≤0.05). The effect of harmaline on food and water intake was significantly attenuated with ketanserin and SB242084 pretreatment (P ≤0.05). These results suggest that there is an interaction between harmaline and 5-HT (via 5-HT 2a and 5-HT 2c receptors) on eating response in chicken. Key words: Harmaline, 5-HT, Eating response, Chicken Introduction content of -carbolines: harmine, harmaline and tetrahydroharmine (THH), collectively -carbolines are a class of alkaloids that known as harmala alkaloids. The have elicited considerable research interest interactions of harmala alkaloids at the 5- (Pfau and Skog, 2004). They are naturally HT, dopamine and benzodiazepine receptors occurring alkaloids that exhibit a wide range are a rather controversial question (Glennon of psychopharmacological effects because of et al ., 2000). Harmaline also induces a shift their binding Archiveto serotonin (5-HT), inof emotional SID reactivity, particularly in benzodiazepine, imidazoline and opiate anxiety displaying anxiogenic and anxiolytic receptors as well as monoamine oxidase actions (Hilber and Chapillon, 2005). It is (MAO) inhibition (Herraiz and Chaparro, thus important to further evaluate the 2005; Herraiz and Chaparro, 2006a, b; potential beneficial or toxic effects of these Herraiz et al ., 2008). In nature, -carboline compounds, using behavioral endpoints of alkaloids are reported to be found in a neural function. number of plants, including Banisteriopsis On the basis of psychopharmacological caapi ( Malpighiaceae ) and Peganum effects of harmaline and its binding to 5-HT harmala L. ( Zygophyllaceae ), their extracts receptors as well as MAO inhibition, we exhibit psychoactive actions mediated hypothesized that 5-HTergic system and/or potentiated by these compounds possibly mediates harmaline signaling in the (Callaway et al ., 2005). The possible use of hypothalamus of birds. Thus, the present P. harmala in modern phyto-indole study was designed for the first time to entheogen preparations is correlated to its investigate whether blocking 5-HT2a and 5- www.SID.ir 94 Iranian Journal of Veterinary Research, Shiraz University, 2013, Vol. 14, No. 2, Pages 94-99 HT 2c receptors can influence harmaline- Orthodontics) trimmed to the exact length of induced feeding and drinking response in the guide cannula was inserted into the guide FD24. cannula while the chicks were not being used for experiments. Lincospectin (Razak, Materials and Methods Tehran, Iran) was applied to the incision to prevent infection. The birds were allowed a Animals minimum of 5 days recovery prior to Day-old Ross 308 broiler cockerels injection. (Eshragh Hatchery, Varamin, Iran) were housed in heated batteries with continuous Experimental procedures lighting until 3 weeks of age. Birds were To determine the possible effects of provided with a mash diet (21% protein and harmaline on food and water intake and its 2,869 kcal/kg of metabolizable energy) and interaction with 5-HTergic system, four water ad libitum . At approximately 2 weeks experiments were conducted. Twenty-four of age, the birds were transferred to birds were used in each experiment (total individual cages. The temperature and used chickens for all experiments was 96). relative humidity of the animal room were Injections were made with a 29-gauge, thin- maintained at 22 ± 1°C and 50%, walled stainless steel injecting cannula respectively, in addition to the continuous which extended 1.0 mm beyond the guide lighting condition. This study was approved cannula. This injecting cannula was by the ethics committee of Faculty of connected through a 60-cm-long PE-20 Veterinary Medicine of Tehran University. tubing to 10-µl Hamilton syringe. Solutions Animal handling and experimental were injected over a period of 60 s. A procedures were performed according to the further 60 s period was allowed to permit the Guide for the Care and Use of Laboratory solution to diffuse from the tip of the Animals published by the US National cannula into the ventricle. All experimental Institutes of Health (NIH Publication procedures were performed during a period NO.85-23, revised 1996). from 10:00 a.m. to 02:00 p.m. Before injection, the birds were removed from their Drugs individual cages, restrained by hand, and Harmaline (a MAO inhibitor), ketanserin then put back into their cages after injection. (5-HT 2a receptor antagonist) and SB242084 Birds were handled and injected daily during (5-HT 2c receptor antagonist) were purchased the 5-day recovery period, in order to from Tocris Cookson Co. (Bristol, UK). All become used to the injection procedure. drugs were dissolved in 1% dimethyl Tubing and syringes were kept in 70% sulfoxide (DMSO). Control group received ethanol, and the glassware was autoclaved to vehicle as control. render materials pyrogen-free. Three hours before the start of the experiments, animals Surgical preparation were deprived of food and water. At 3 weeksArchive of age, broilers were Immediatelyof SIDafter injection, the birds were anesthetized with sodium pentobarbital returned to their cages. Fresh food and water (Sagatal, Rhone Merieux) (25 mg/kg body were supplied at the time of injection and weight, iv) and a 23-gauge thin-walled cumulative food (g) and water (g) intake stainless steel guide cannula was were recorded at 15, 30, 45, 60, 90, 120, 150 stereotaxically implanted into the right and 180 min after injection. Placement of lateral ventricle and the stereotaxic the guide cannula into the ventricle was coordinates were AP = 6.7, L = 0.7, H = 3.5- verified by the presence of cerebrospinal 4 mm below the duramater with the head fluid and intracerebroventricular injection of oriented. The cannula was secured with methylene blue followed by slicing the three stainless steel screws placed in the frozen brain tissue at the end of the calvaria surrounding each guide cannula, experiments. then acrylic dental cement (Pars acryl) was In this study, experiment 1 was applied to the screws and guide cannula. An performed to examine the effect of ICV orthodontic #014 wire (American injections of harmaline on food and water www.SID.ir 95 Iranian Journal of Veterinary Research, Shiraz University, 2013, Vol. 14, No. 2, Pages 94-99 intake in FD24 (n = 6 for each group) following experiments because it was found chickens. The birds received 25, 50 and 100 to induce potent decrease in food intake and µg harmaline in 10-µl vehicle. Control increase in water intake in FD24 animals group was injected with 10 µl of vehicle. without affecting other non-ingestive In experiment 2 and 3 each bird received behavioral parameters. two injections with 15 min interval and fresh In experiment 2, intracerebroventricular food and water were supplied at the time of injection of 100 g harmaline alone second injection and cumulative food (g) decreased food intake and increased water and water (ml) intake were recorded at 15, intake (P ≤0.05). In addition, the effect of 30, 45, 60, 90, 120, 150 and 180 min after harmaline on cumulative food and water the second injection. intake was significantly decreased by 10 µg On the other hand, in experiment 2, the ketanserin pretreatment (P ≤0.05) (Figs. 3 first injection consisted of either 0 or 10 µg and 4). ketanserin in 5-µl vehicle. The second injection consisted of either 0 or 100 µg Vehicle Harmaline (25 µg) harmaline in 5-µl vehicle. Harmaline (50 µg) Harmaline (100 µg ) 80 Experiment 3 was conducted parallel to a 70 a a the experiment 2, except that the chicks a 60 a received 0 or 3 µg SB242084 instead of a 50 a ketanserin. a b 40 a b The dosages of drugs were selected a b a 30 a b c based on previous studies (Von Meyenburg (g) intake Food b c a a c et al ., 2003; Medeiros et al ., 2005; Bungo et 20 b c a a b c 10 al ., 2008) and preliminary experiments. b c c c During the injection of higher doses of 0 15 30 45 60 90 120 150 180 harmaline, the birds were very active, Time (min) excited and sometimes convulsive. Fig. 1: Effect of intracerebroventricular Statistical analysis injection of different doses of harmaline on food intake in 24-h food-deprived (FD24) Cumulative food and water intake were chickens. Vehicle: 1% DMSO: Control. Data analysed by one-way analysis of variance are presented as mean ± SEM.
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