Effects of intracerebroventricular injection of muscimol or GABA on operant feeding in pigs

Young pigs were prepared with lateral intracerebroventricular (ICV) cannulae. They were housed individually in cages fitted with operant panels and could obtain food and water ad lib. The GABA-A receptor agonist muscimol (25-200 nmol) ICV produced an increase in food intake in which the dose-response relation was most obvious 30-60 min after dosing. The 25-nmol dose had no effect on feeding. However, muscimol (50 nmol) caused a significant increase in feeding (p less than 0.01) during the first 30 min after injection, while the 100- and 200-nmol doses increased food intake (p less than 0.01) during the first 60 min. The effect of muscimol (100 nmol) on food intake was completely abolished by the simultaneous administration of the GABA-A receptor antagonist bicuculline (100 nmol). GABA (40-1600 nmol) ICV also produced a dose-related increase in food intake (p less than 0.01) in the 15 min after injection. Only doses of 800 nmol and above were effective. The effects of GABA (1600 nmol) were completely abolished by the simultaneous administration of bicuculline (50 nmol). Neither muscimol nor GABA influenced food intake for the 24-hr time period or water intake during any time period. The results indicate that stimulation of central GABA-A receptors induces operant feeding in the satiated pig.     

GABA(A) receptor activation in the lateral parabrachial nucleus induces water and hypertonic NaCl intake

Inhibitory serotonergic and cholecystokinergic mechanisms in the lateral parabrachial nucleus and central GABAergic mechanisms are involved in the regulation of water and NaCl intake. In the present study we investigated if the GABA(A) receptors in the lateral parabrachial nucleus are involved in the control of water, NaCl and food intake in rats. Male Holtzman rats with stainless steel cannulas implanted bilaterally into the lateral parabrachial nucleus were used. Bilateral injections of muscimol (0.2 nmol/0.2 microl) into the lateral parabrachial nucleus strongly increased 0.3 M NaCl (20.3+/-7.2 vs. saline: 2.6+/-0.9 ml/180 min) without changing water intake induced by the treatment with the diuretic furosemide combined with low dose of the angiotensin converting enzyme inhibitor captopril s.c. In euhydrated and satiated rats, bilateral lateral parabrachial nucleus injections of muscimol (0.2 and 0.5 nmol/0.2 microl) induced 0.3 M NaCl intake (12.1+/-6.5 and 32.5+/-7.3 ml/180 min, respectively, vs. saline: 0.4+/-0.2 ml/180 min) and water intake (5.2+/-2.0 and 7.6+/-2.8 ml/180 min, respectively, vs. saline: 0.8+/-0.4 ml/180 min), but no food intake (2+/-0.4 g/240 min vs. saline: 1+/-0.3 g/240 min). Bilateral lateral parabrachial nucleus injections of the GABA(A) antagonist bicuculline (1.6 nmol/0.2 microl) abolished the effects of muscimol (0.5 nmol/0.2 microl) on 0.3 M NaCl and water intake. Muscimol (0.5 nmol/0.2 microl) into the lateral parabrachial nucleus also induced a slight ingestion of water (4.2+/-1.6 ml/240 min vs. saline: 1.1+/-0.3 ml/240 min) when only water was available, a long lasting (for at least 2 h) increase on mean arterial pressure (14+/-4 mm Hg, vs. saline: -1+/-1 mm Hg) and only a tendency to increase urinary volume and Na+ and K+ renal excretion. Therefore the activation of GABA(A) receptors in the lateral parabrachial nucleus induces strong NaCl intake, a small ingestion of water and pressor responses, without changes on food intake.     

Interaction between nociceptin/orphanin FQ (N/OFQ) and GABA in response to feeding

Nociceptin/orphanin FQ (N/OFQ) and gamma aminobutyric acid (GABA) agonists have been shown to increase feed intake in mammals and birds. In this study, the effect of intracerebroventricular (icv) injection of the potent NOP receptor agonists Nociceptin (1-13) NH(2), the GABA(A) receptor antagonist bicuculline, and the GABA(A) receptor agonist muscimol on feed intake in cockerels was investigated. The icv injection of N/OFQ and muscimol increases feed intake. The effect of N/OFQ on feed intake was strongly blocked by the injection of bicuculline whereas the effect of muscimol was stimulated by N/OFQ. These results suggest that N/OFQ may act at GABA(A) receptors or increases overflow of GABA in the brain of chickens to stimulate feeding.     

Cannabinoid agonist, CP 55,940, facilitates intake of palatable foods when injected into the hindbrain

Cannabinoids have been shown to influence food intake, and until recently, the neural pathways mediating these effects have remained obscure. It has been previously shown that intracerebroventricular injection of delta-9-tetrahydrocannabinol (Δ⁹-THC) causes increased consumption of palatable foods in rats, and we postulated the involvement of the hindbrain in this cannabinoid-induced food intake. Cannulated rats (both female and male groups) trained to consume sweetened condensed milk received either lateral or fourth ventricle injections of CP 55,940 and were presented with sweetened condensed milk 15 min after injection. Rats were injected over a range of doses between 100 pg and 10 μg per rat. Milk intake was recorded for a total of 3 h. Lateral ventricle injection of CP 55,940 increased milk intake at doses in the microgram range. However, CP 55,940 was effective in increasing food intake at nanogram doses when injected into the fourth ventricle. Finally, male rats appeared to be more sensitive to CP 55,940 than female rats inasmuch as milk consumption was increased at the 1 ng dose in male rats, whereas only the 10 ng dose was effective in females. These results indicate that CP 55,940 may act in the hindbrain to influence feeding behavior in rats.     

The differential effects of intravenously administered 8-OH-DPAT on operant food intake in satiated and food-deprived pigs are mediated by central 5-HT(1A) receptors

It has previously been shown that the intravenous administration of the 5-HT(1A) receptor agonist, 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT), increases food intake in satiated pigs and decreases food intake in fasted pigs. The present experiments were conducted to investigate the effects of central administration of the 5-HT(1A) receptor antagonist, N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]-N-2-pyridinyl-cyclohexane carbox-amide maleate (WAY 100635), on the stimulant and depressant effects of 8-OH-DPAT on operant food intake in satiated and hungry pigs. In Experiment 1, 8-OH-DPAT (25 microg/kg) produced an increase in operant feeding during the first 30 min following intravenous administration to satiated pigs. The 8-OH-DPAT-induced hyperphagia was completely abolished by pretreatment with WAY 100635 (0.3 mg) administered by intracerebroventricular injection. In Experiment 2, 8-OH-DPAT (25 microg/kg) administered intravenously 15 min prior to the onset of feeding in pigs that had been fasted for 22.5 h produced a decrease in operant food intake, which was most apparent during the first 30 min of the feeding period. The hypophagic effect was completely abolished by pretreatment with WAY 100635 (0.3 mg icv) administered 30 min before the start of the feeding period. In both experiments, WAY 100635 (0.3 mg icv) did not have any significant effects on feeding. The results of the present study extend previous results in the pig and show that both the hyperphagic and the hypophagic effects of 8-OH-DPAT in satiated and fasted pigs, respectively, are mediated by central 5-HT(1A) receptors.     

Effect of the 5-HT(1A) receptor agonist 8-OH-DPAT on food and water intake in chickens

The effects of the 5-HT(1A) receptor agonist 8-OH-DPAT on food and water intake in 16-week-old male chickens were investigated. Injection of 25 or 50 microg/kg of 8-OH-DPAT 15 min before refeeding starved animals (starved-refed) produced a decrease in food intake 1 h after the start of refeeding. No effect was observed in water intake. The injection of 25 or 50 microg/kg of the 8-OH-DPAT 60 min after the start of refeeding (fed) produced increased food intake, but no effect was observed on water intake. The agonist 8-OH-DPAT (50 microg/kg) injected in water-deprived chickens 15 min before water presentation produced a rapid increase in water intake and an increase in food intake 90 min after the presentation of water. The effect on food intake was mainly apparent 60-90 min after injection. However, when the chickens were water-deprived, the intravenous administration of 8-OH-DPAT 15 min before the presentation of water produced an increase in water intake only 15 min after the start of the experiment. The results show that the effect on food intake of the agonist 8-OH-DPAT in chickens was similar to that observed in mammals. Also, the results show that the agonist-induced increase in water intake may act via a different mechanism. The results show that the 8-OH-DPAT, as in mammals, has a complex effect on food and water intake in chickens and that further works need to be carried out to understand the mechanisms involved in the food and water intake using different animal models.     

GABAergic control of food intake in the meat-type chickens

This study examined the effects of intracerebroventricular injections of gamma-aminobutyric acid (GABA) agonists on short-term food intake in meat-type cockerels. In Experiment 1, birds were injected with various doses of muscimol, a GABA(A) agonist. In Experiment 2, the birds received bicuculline, a GABA(A) antagonist, prior to injection of muscimol. In Experiment 3, the effect of varying doses of baclofen, a GABA(B) agonist, on food intake was determined. The intracerebroventricular injection of muscimol caused a dose-dependent increase in food intake. This effect was significantly attenuated by pretreatment with bicuculline. Food intake was not affected by the intracerebroventricular injection of baclofen. These results suggest that GABA acts within the brain of broilers at a GABA(A), but not GABA(B), receptor to increase voluntary food intake.     

Intraperitoneal and intracranial cholecystokinin depress operant responding for food.

Intraperitoneal injection of 20 I.D.U./kg of cholecystokinin (CCK), a gut hormone, depressed food intake in female rats as measured by an operant response for food. The effect was maximal thirty min after the injection and lasted 60–90 min. Intracranial injection of 0.05, 0.10, or 0.20 I.D.U. of CCK into the lateral ventricle of female rats, produced a dose-related depression of food-rewarded lever pressing. In this case, however, the effect was not observed during the first thirty min, but became apparent thereafter. The results are discussed in terms of possible receptors for CCK within the brain.     

Centrally administered ghrelin potently inhibits water intake induced by angiotensin II and hypovolemia in rats

Ghrelin is a potent, centrally acting orexigenic hormone. Recently, we showed that centrally administered ghrelin is a potent antidipsogenic hormone in 24-h water deprived rats. In this study, we examined the effect of intracerebroventricular (icv) injection of ghrelin on angiotensin II (AII)-induced water intake in rats. We also examined the effects of icv injection of ghrelin on drinking induced by intraperitoneal injection of an isotonic polyethylene glycol (PEG) solution that causes isotonic hypovolemia. Water intake induced by the icv injection of AII or ip injection of PEG was significantly reduced after icv injection of ghrelin, although food intake was stimulated by the hormone. The drinking induced by AII was also inhibited by the icv administration of 4α-phorbol 12, 13-didecanoate, an agonist of the osmosensitive TRPV4 channel. This study showed that ghrelin is a potent antidipsogenic peptide by antagonizing general dipsogenic mechanisms including those activated by AII and hypovolemia in rats.     

中枢CRH在大鼠应激性体温升高和LPS性发热机制中的作用

目的:进一步观察中枢促肾上腺皮质激素释放激素(CRH)在大鼠应激性体温升高和脂多糖(LPS)性发热中枢机制中的作用。方法:第三脑室微量注射CRH受体拮抗剂α-helicalCRH(9-41)和LPS,测定大鼠结肠温度及脑腹中隔区精氨酸加压素(AVP)含量。结果:生理盐水对照组大鼠体温明显升高,最大升幅为(0.88±0.31)℃。第三脑室注射CRH受体拮抗剂α-helicalCRH(9-41)10min后再注射生理盐水组,90min内大鼠结肠温度未见明显波动,90min后体温开始上升,1.5h体温反应指数(TRI_1.5)明显低于生理盐水对照组,而脑腹中隔区AVP含量和TRI_3.5与对照组比较均没有明显差别。第三脑室注射300ng的LPS引起大鼠结肠温度双相性升高,其TRI_3.5明显高于生理盐水对照组。事先向第三脑室注射α-helicalCRH(9-41)(5μg)再注射LPS组,TRI_3.5明显高于LPS组,而脑腹中隔区AVP含量明显低于LPS组。结论:CRH介导应激诱导的早期体温升高。CRH可能通过诱导脑腹中隔区AVP的生成限制大鼠LPS性发热。在大鼠LPS性发热中,CRH可能是一种双相作用分子,一方面本身介导发热体温升高,另一方面又诱生发热体温负调节介质而限制发热体温的升高。     

Electrical activity and feeding correlates of intracranial hypothalamic injection of GABA, muscimol and picrotoxin in the rats

Assemblies of electrodes and a cannula were stereotaxically implanted in the ventromedial (VMH), lateral (LHA) and paraventricular (PVH) hypothalamic areas in male albino rats. Electrical activity of these regions was recorded electrographically before and following intracranial injection (ICI) of GABA, muscimol and picrotoxin. In another set of animals, food intake and water intake were also measured. The activity of the ventromedial hypothalamus changed from slow to fast after ICI of GABA and picrotoxin and fast to slow after muscimol. The activity of the lateral hypothalamus changed from slow to fast with ICI of muscimol and picrotoxin and from fast to slow with GABA, while that of the paraventricular hypothalamic nucleus changed from slow to fast with ICI GABA and fast to slow with muscimol and picrotoxin. ICI of GABA into VMH and LHA and muscimol in VMH, LHA and PVH caused a decrease in food intake. Water intake was also decreased after ICI of GABA in PVH and muscimol in LHA and PVH. On the opposite picrotoxin increased food intake in VMA and LHA and water intake in PVH. The possible interaction of GABAergic drugs with the areas of the brain controlling feeding and drinking is being discussed.     

Increased food intake and carbohydrate preference in the rat following treatment with the serotonin antagonist metergoline

The effect on feeding of antagonism of the serotonergic system was studied. Intraperitoneal injection of one of 8 doses of the serotonin antagonist metergoline (MET) (0.06-1.5 mg.kg-1) given prior to a 3 h food deprivation produced a significant dose-related increase in intake of a standard laboratory diet throughout a 2 h measurement period. A 1 mg/kg dose of MET administered prior to one of 3 isocaloric powdered diets a medium carbohydrate, a high carbohydrate, and a non-carbohydrate diet - produced a significant increase in the intake by rats of all 3 diets in the first hour after food presentation. However, in the second hour MET continued to increase consumption of the high carbohydrate diet, whereas intakes of the medium carbohydrate and non-carbohydrate diets were no longer affected by MET treatment. These data suggest that the serotonergic system is a tonic inhibitor of food intake and of carbohydrate preference.     

Feeding and locomotion responses to centrally injected nociceptin/orphanin FQ in chicks

Nociceptin/orphanin FQ (N/OFQ), the endogenous ligand of the opioid receptor-like receptor or nociceptin receptor (NOP), has been shown to induce feeding, locomotion, anti-stress and anxiolytic effects in rodents after central nervous system injection. In this study, the effect of intracerebroventricular (icv) injection of N/OFQ on feeding and locomotion behavior was evaluated in male broiler-type chickens. The icv injection of N/OFQ caused a moderate but significant increase in feed intake similar to the classical opioid peptides in rats. It also increased feed pecking frequency and feeding time 1 h after injection. Stepping, wing flapping and preening were not affected by N/OFQ. These results suggest that N/OFQ can act within the central nervous system of chickens to increase feed intake.     

Activation of brain somatostatin 2 receptors stimulates feeding in mice: Analysis of food intake microstructure

We recently reported that the oligosomatostatin receptor agonist, ODT8-SST increases food intake in rats via the somatostatin 2 receptor (sst₂). We characterized ingestive behavior following intracerebroventricular (icv) injection of a selective sst₂ agonist in freely fed mice during the light phase. The sst₂ agonist (0.01, 0.03, 0.1, 0.3 or 1 μg/mouse) injected icv under short inhalation anesthesia dose-dependently increased cumulative light phase food intake over 4 h compared to vehicle with a 3.1-times increase at 1 μg/mouse (p < 0.05). Likewise, the sst_2,3,5 agonist octreotide (0.3 or 1 μg/mouse) dose-dependently increased 4-h food intake, whereas selective sst₁ or sst₄ agonists at 1 μg/mouse did not. In vehicle-treated mice, high fat diet increased caloric intake/4 h by 2.8-times compared to regular diet (p < 0.05) and values were further increased 1.4-times/4 h by the sst₂ agonist. Automated continuous assessment of food intake established a 6.6-times higher food intake during the dark phase due to increased number of meals, meal size, meal duration and rate of ingestion compared to non-treated mice during the light phase. During the first 4 h post icv sst₂ agonist injection, mice had a 57% increase in number of meals with a 60% higher rate of ingestion, and a 61% reduction in inter-meal intervals, whereas meal sizes were not altered compared to vehicle. These data indicate that the activation of brain sst₂ receptors potently stimulates the light phase ingestive behavior under basal or high fat diet-stimulated conditions in mice. The shortened inter-meal interval suggests an inhibitory effect of the sst₂ agonist on “satiety”, whereas “satiation” is not altered as indicated by normal meal size.     

Induction of food intake by a GABAergic mechanism in the turkey.

The effect of GABAergic influences on food and water intake in Large White turkeys was investigated. In Experiment 1, food and water intake were monitored in hens injected intracerebroventricularly (ICV) with varying doses of muscimol, a GABA agonist. In Experiment 2, hens were pretreated with ICV injections of picrotoxin, a GABA antagonist, prior to the injection of muscimol. In Experiment 3, the effect of ICV injections of muscimol on water intake was determined in hens not allowed access to food. The ICV injection of muscimol caused a dose-dependent increase in food intake with 75 nmole being the most efficacious dose. Water intake was stimulated following the injection of muscimol when food and water were available ad lib. However, water intake was not affected by the ICV injection of muscimol if food was not simultaneously available, indicating that the increased water intake was a result of increased food intake. The effect of muscimol on food intake was significantly attenuated by pretreatment with picrotoxin. It appears, therefore, that GABA acts within the brain of the turkey to increase food, but not water, intake.     

An fMRI study of acupuncture using independent component analysis

While the hypothalamus has been implicated in the regulation of energy balance, the central mechanisms and neural circuit that coordinate the feeding response to energy deficit have not been fully clarified. To better understand the role of the hypothalamus in mediating hyperphagic responses to food deprivation or glucoprivation, we examined the feeding responses in rats in which the medial hypothalamus (MH) was isolated from the rest of the brain. The isolation of the MH was performed with a Halasz's knife cut, and experiments were performed 7 days after the operation. Food consumption between 9:00 a.m. and 11:00 a.m. in rats which had been fasted overnight was significantly increased compared to that in rats which had access to food ad libitum before the measurement in both the sham and MH-isolated groups, and the absolute values of food consumption in fasted rats were not significantly different between the groups. On the other hand, while an injection of 2-deoxy-d-glucose, which blocks glucose utilization, significantly increased food consumption for 2 h after injection compared to a saline injection in the sham group, it did not increase food intake compared to saline injection in the MH-isolated groups. Thus, it is demonstrated that glucoprivation is not an effective stimulus to induce feeding in MH-isolated rats.     

The role of GABAergic system on the inhibitory effect of ghrelin on food intake in neonatal chicks

Ghrelin is a gut-brain peptide that has a stimulatory effect on food intake in mammals. In contrast, this peptide decreases food intake in neonatal chicks when injected intracerebroventricularly (ICV). In mammals, neuropeptide Y (NPY) mediates the orexigenic effect of ghrelin whereas in chicks it appears that corticotrophin releasing factor (CRF) is partially involved in the inhibitory effect of ghrelin on food intake. Gamma aminobutyric acid (GABA) has a stimulatory effect on food intake in mammals and birds. In this study we investigated whether the anorectic effect of ghrelin is mediated by the GABAergic system. In Experiment 1, 3 h-fasted chicks were given an ICV injection of chicken ghrelin and picrotoxin, a GABA_A receptors antagonist. Picrotoxin decreased food intake compared to the control chicks indicating a stimulatory effect of GABA_A receptors on food intake. However, picrotoxin did not alter the inhibitory effect of ghrelin on food intake. In Experiment 2, THIP hydrochloride, a GABA_A receptor agonist, was used in place of picrotoxin. THIP hydrochloride appeared to partially attenuate the decrease in food intake induced by ghrelin at 30 min postinjection. In Experiment 3, the effect of ICV injection of chicken ghrelin on gene expression of glutamate decarboxylase (GAD)₁ and GAD₂, GABA synthesis enzymes in the brain stem including hypothalamus, was investigated. The ICV injection of chicken ghrelin significantly reduced GAD₂ gene expression. These findings suggest that ghrelin may decrease food intake in neonatal chicks by reducing GABA synthesis and thereby GABA release within brain feeding centers.     

The Injection of Hypocretin-1 into the Nucleus Pontis Oralis Induces either Active Sleep or Wakefulness Depending on the Behavioral State when it is Administered

Study Objectives:

We previously reported that the microinjection of hypocretin (orexin) into the nucleus pontis oralis (NPO) induces a behavioral state that is comparable to naturally occurring active (rapid eye movement) sleep. However, other laboratories have found that wakefulness occurs following injections of hypocretin into the NPO. The present study tested the hypothesis that the discrepancy in behavioral state responses to hypocretin injections is due to the fact that hypocretin was not administered during the same states of sleep or wakefulness.

Design:

Adult cats were implanted with electrodes to record sleep and waking states. Hypocretin-1 (0.25 μL, 500mM) was microinjected into the NPO while the animals were awake or in quiet (non-rapid eye movement) sleep.

Measurements and Results:

When hyprocretin-1 was microinjected into the NPO during quiet sleep, active sleep occurred with a short latency. In addition, there was a significant increase in the time spent in active sleep and in the number of episodes of this state. On the other hand, the injection of hyprocretin-1 during wakefulness resulted not only in a significant increase in wakefulness, but also in a decrease in the percentage and frequency of episodes of active sleep.

Conclusions:

The present data demonstrate that the behavioral state of the animal dictates whether active sleep or wakefulness is induced following the injection of hypocretin. Therefore, we suggest that hypocretin-1 enhances ongoing states of wakefulness and their accompanying patterns of physiologic activity and that hypocretin-1 is also capable of promoting active sleep and the changes in various processes that occur during this state.

Citation:

Xi M; Chase MH. The injection of hypocretin-1 into the nucleus pontis oralis induces either active sleep or wakefulness depending on the behavioral state when it is administered. SLEEP 2010;33(9):1236-1243.     

Losartan blocks drinking and cFos expression induced by central ornithine vasotocin in rats

We previously reported that an intracerebroventricular (icv) injection of the oxytocin receptor antagonist ornithine vasotocin (OVT) caused water and saline intakes, a pressor response, and Fos-like immunoreactivity (Fos-IR) in the median preoptic nucleus of the rat brain. In the present report, rats receiving an icv injection of isotonic saline vehicle followed by an icv injection of 10 microg of OVT 20 min later drank 5.5+/-1.1 ml of total water and saline intake in 60 min after the OVT; rats receiving 10 microg of losartan before the OVT drank only 0.9+/-0.3 ml of total fluid. In a separate study, rats were treated as above except that they were not allowed to drink and were perfused for analysis of Fos-IR in the median preoptic nucleus at 90 min. Fos-IR in the dorsal part of the median preoptic nucleus was significantly suppressed from 2.69+/-0.57 cells per 10,000 square mum in vehicle-treated rats to 0.89+/-0.20 in losartan-treated rats. Losartan alone had no effect on Fos-IR. Losartan did not reduce intake of saccharin in a dessert test. This suggests that the OVT-induced drinking may result from an activation or disinhibition of angiotensin type AT1 receptors in the median preoptic nucleus.     

Estradiol decreases the orexigenic effect of melanin-concentrating hormone in ovariectomized rats

Estradiol exerts an inhibitory effect on food intake via interactions with anorexigenic peptides, like cholecystokinin, that function to decrease meal size. It is currently unknown whether estradiol also interacts with orexigenic compounds implicated in the physiological control of food intake. Thus, the primary goal of this study was to determine whether estradiol decreases the orexigenic effect of melanin-concentrating hormone (MCH), a neuropeptide that, like estradiol, appears to influence food intake by selectively affecting the controls of meal size. Food and water intake were monitored following lateral ventricular (icv) infusions of 5 mug MCH or saline vehicle in oil- and estradiol-treated ovariectomized rats. MCH increased food intake throughout the first 4 h of the dark phase in oil-treated rats, but only for the last 2 h of the same 4-h interval in estradiol-treated rats. As a result, the orexigenic effect of MCH was significantly lower in estradiol-treated rats, relative to oil-treated rats. During this interval of MCH-stimulated feeding, a prandial increase in water intake was not observed in either oil- or estradiol-treated rats. We conclude that estradiol decreases the orexigenic effect of MCH in ovariectomized rats.