Lateral hypothalamic injection of bombesin decreases food intake in rats

STUCKEY, J. A. AND J. GIBBS. Lateral hypothalamic injection of bombesin decreases food intake in rats. BRAIN RES. BULL. 8(6)617–621, 1982.—The effect of lateral hypothalamic injections of bombesin on feeding behavior was examined. Rats equipped with stainless steel cannulas directed toward the lateral hypothalamus received bilateral injections of bombesin prior to access to a liquid test diet after a 3 hr food deprivation. Bombesin in doses of 5 ng, 50 ng and 100 ng produced significant reductions in the size of the first meal. Injection of 50 ng of the biologically weak analogue [D-Trp⁸] bombesin had no effect. Injection of 5 ng or 50 ng of bombesin had no effect on deprivation-induced water intake, and injection of 50 ng of bombesin had no effect on body temperature. The food and water intake data and direct quantitative behavioral measures indicated that lateral hypothalamic injections of bombesin specifically reduced food intake. The structure-activity relationship for this effect was similar to those for other actions of bombesin. A bombesin-like peptide in the lateral hypothalamus or its receptors may play a role in postprandial satiety.     

Xenin-a novel suppressor of food intake in rats

Peptides related to the amphibian octapeptide xenopsin are present in various locations in mammalians, such as the gastrointestinal mucosa or brain tissue. In the gastrointestinal tract, xenopsin-related peptides induce partially neurogenic contractions of the colon in humans. In brain, however, their function is not known. Structural similarities of xenopsin-related peptides with neurotensin, a known modulator of ingestive behavior, suggest a possible role in feeding regulation. Therefore, we examined the effect of xenin, a recently identified xenopsin-related pentacosa peptide, on feeding behavior of fasted rats. Male Wistar rats (n=12) were intracerebroventricularly (i.c.v.) injected with either saline (10 μl) or xenin at 0.5, 1.5, 5 or 15 μg dissolved in an identical volume of 10 μl, respectively. In further experiments, xenin 15 μg/0.5 μl or 0.5 μl saline were injected into the lateral hypothalamus (LH). After injections, food intake (g), percentage of time spent with feeding (%) and prandial water intake (ml) were subsequently recorded for 2 h. After i.c.v. injection of 15 μg of xenin 1-h food intake was significantly reduced by 42% and 2-h food intake was diminished by 25%, respectively, compared to saline injection (p<0.01). This reduction of food intake was paralleled by a significant decrease of time spent with feeding by 41% (after 1 h) or 23% (after 2 h). The xenin-induced suppression of feeding behavior was dose-dependent. Thus, the minimal effective dose of xenin was 1.5 μg, while the dose of 0.5 μg was ineffective. Prandial water intake was significantly reduced only by the highest dose of xenin. Following injection of 15 μg of xenin into the lateral hypothalamus food intake was not different from control experiments. These data demonstrate a potent feeding suppressive action of xenin following intracerebroventricularly injection but not injection into the lateral hypothalamus suggesting a possible role of xenin in the central control of feeding termination and satiety.     

Neuronal histamine in the hypothalamus suppresses food intake in rats

Using probes to manipulate hypothalamic neuronal histamine, we report here that changes in neuronal histamine modulate physiological feeding behavior in rats. Infusion of α-fluoromethylhistidine (FMH), a “suicide” inhibitor of histidine decar☐ylase (HDC), into the third cerebroventricular induced feeding in the early light phase when the histamine synthesis was most accelerated. FMH at an optimum 2.24 μmol dose elicited feeding in 100% of rats. Treatment of FMH specifically and selectively decreased concentration of histamine without affecting concentrations of catecholamines in the hypothalamus. Immediately before the dark phase, when the histamine synthesis was normally lower, FMH infusion did not affect feeding-related parameters such as meal size, meal duration or latency to eat. Conversely, thioperamide, which facilitates both synthesis and release of neuronal histamine by blocking presynaptic autoinhibitory H₃ receptors, significantly decreased food intake after infusion of a 100-nmol dose into the third cerebroventricle. The effect of thioperamide was abolished with i.p. injection of 26 μmol/kg chlorpheniramine, an H₁ antagonist. FMH at 224 nmol was microinfused bilaterally into the feeding-related nuclei in the hypothalamus. The ventromedial nucleus (VMH) and the paraventricular nucleus (PVN), but not the lateral hypothalamus, the dorsomedial hypothalamus or the preoptic anterior hypothalamus were identified as the active sites for the modulation. Neuronal histamine may convey suppressive signals of food intake through H₁ receptors in the VMH and the PVN with diurnal fluctuation.     

Role of glucagon in the control of food intake in zucker obese and lean rats

Although there is strong evidence for glucagon's role in the control of food intake, the essentiality of this role remains in question. In several experiments the feeding responses to glucagon and glucagon antisera were investigated in both Zucker and Sprague-Dawley rats. Intraperitoneal injection of 400 micrograms/kg glucagon decreased 30-min food intake 18% (p less than 0.01) in Zucker lean rats and increased 30-min food intake 16% (NS) in Zucker obese rats, suggesting obese rats are less sensitive. In Sprague-Dawley rats the same dose decreased first meal size 28% (p less than 0.01), indicating that they were more sensitive than Zucker lean rats. Intraperitoneal injection of 400 micrograms/kg glucagon increased plasma glucagon concentrations in the vena cava and the tail vein 150-fold and 10-fold, thus, superphysiological doses may be required to elicit satiety. In contrast, administration of a glucagon antisera increased food intake of Zucker rats for up to 6 hr and increased meal size for 5 hr. The findings suggest that glucagon's role in control of food intake in Zucker obese and lean rats is similar, but the superphysiological glucagon changes which occur with exogenous administration indicate that glucagon may only indirectly elicit satiety.     

Visceral factors in the control of food intake

Some years ago, we reported that the increased blood intake of hypoglycemic rats was inhibited by the intravenous infusion of fructose, a sugar that cannot cross the blood-brain barrier and nourish cerebral chemoreceptors. More recent experiments therefore have focused on visceral factors in the control of food intake. Three observations have been emphasized in this review. First, we found that gastric emptying was increased during insulin-induced hypoglycemia, and that this effect also was eliminated by administration of fructose. Hepatic vagotomy abolished both this effect of fructose on gastric emptying and its effect on food intake. Second, we found that in rats with severe diabetes, the rate of gastric emptying did decrease in proportion to increasing concentration of an administered glucose load, as it does in intact rats, but calories emptied more rapidly than normal regardless of the concentration of the load. Third, we found that rats with varying degrees of streptozotocin-induced damage to the pancreas ate more food than intact rats did after an overnight fast, and that individual intakes were proportional to the induced glucose intolerance. The increased eating took the form of shorter intermeal intervals, as if the initial postfast meal did not remain satiating for a normal amount of time. These and other findings suggest that food intake is controlled in part by satiety signals apparently related to the delivery of utilizable calories plus insulin to the liver. These signals also seem to affect gastric emptying and thereby might influence other satiety signals related to gastric distention.     

Commissural nucleus of the solitary tract lesions reduce food intake and body weight gain in rats

This study investigated the effects of an electrolytic lesion of the commissural subnucleus of the nucleus of the solitary tract (commNTS) on body weight, daily food and water intake, and plasma glucose and insulin in rats. In the first 6 days following brain surgery, commNTS lesioned rats reduced daily food intake by 80% compared to rats with sham lesions. After this period rats with lesions of commNTS started recovering food intake, but intake remained significantly reduced until the 12th day after surgery. A reduction in body weight was observed 4 days after surgery and reached a maximum on the 12th day. After this, a partial recovery of body weight was observed, but weight remained significantly reduced compared to weights of rats with sham lesions through the conclusion of the study. Food intake and body weight gain in other rats with partial lesions of the commNTS or with lesions outside the commNTS did not differ from rats with sham lesions with regard to those variables. Daily water intake and plasma glucose and insulin were not changed by the commNTS lesions. These results suggest that commNTS is involved with mechanisms that control food intake and body weight in rats.     

Behavioural, biochemical and histological effects of AF64A following injection into the third ventricle of the mouse.

Behavioural, biochemical and histological effects were assessed following AF64A injected into the third ventricle of female NMRI mice. Doses from 3 to 7 nmol produced significant changes in behaviour, causing hyperactivity, reduced hole-board exploration, rotational behaviour in a symmetrical Y-maze corresponding to a loss of alternation, abnormal behaviour in a plus-maze task of fear/anxiety with markedly increased exploration of the open arms and finally deficits in passive avoidance responding and spatial orientation in a Morris-type water maze. In this latter test, a cue learning deficit was noted for the two highest doses only. No histological changes of consequence were observed up to 5 nmol. Beyond this dose, at 6 and particularly 7 nmol, necrosis of parts of the hippocampus and septum was apparent. ChAT and AChE activity were decreased in the hippocampus but not in the cortex although the decreases were smaller than generally reported for AF64A-treated rats. ChAT and AChE reductions correlated highly with hyperactivity in the open-field and to a lesser extent, with spatial learning deficits. Monoaminergic activity was also affected in the hippocampus, but not in the cortex, at 4 nmol and above. NE and particularly 5-HT and 5-HIAA levels were reduced although the rate of 5-HT turnover was unaltered. A highly significant correlation was obtained between 5-HT effects and the increased open arm exploration in the plus-maze task of fear/anxiety. The behavioural effects and biochemical changes lasted at least 8-9 weeks postop.     

Comparative effects of caerulein on food intake and pancreatic secretion in dogs

We compared effects of the CCK analog caerulein on feeding and pancreatic secretion. Nine fasted mongrel dogs with gastric and pancreatic fistulas received scalar doses of caerulein (0, 6.25, 12.5, 25, 50, 100, 200, 400 pmol/kg-hr, each for 30 min). The D50 dose for stimulation of pancreatic secretion was 15 pmol/kg-hr. Effects of intravenous caerulein (0 to 800 pmol/kg-hr; 15 min before and during a 45 min test meal) on food intake were examined in 8 beagles under 4 feeding conditions: 1 meal/day (22 hr fast), 2 meals/day (4 hr fast), 2 meals/day (19 hr fast), and after ad lib access to food followed by a 4 hr fast. The lowest doses that inhibited feeding were: 400 pmol/kg-hr for feeding condition, 200 pmol/kg-hr for and, and 150 pmol/kg-hr for. We conclude: the potency of caerulein for inhibition of food intake is dependent upon feeding condition; these results do not support a role for CCK as a satiety hormone, since the lowest dose of caerulein for inhibition of feeding was 10 times larger than the D50 dose of caerulein for stimulation of pancreatic secretion.     

Neuropeptide Y and food intake in fasted rats: effect of naloxone and site of action

Central administration of neuropeptide Y (NPY) induces food intake in freely feeding animals and this effect is mediated by hypothalamic sites. Little is known, however, about the effect of NPY on food intake and site of action in food-deprived animals. To examine this further, 24-h fasted rats received injections of saline or NPY into the lateral cerebral ventricle (10 micrograms/10 microliters; n = 8) or into the lateral (LH) or ventromedial hypothalamus (VMH) (1 microgram/0.5 microliters; n = 44). In addition, intracerebroventricular (i.c.v.) injections of NPY were carried out with or without i.c.v. naloxone (25 micrograms), a specific opioid receptor antagonist. During the first 40 min food intake was not different with or without NPY. After 60 and 120 min, food intake was 5.9 +/- 0.4 g and 8.3 +/- 0.6 g with i.c.v. saline which was significantly augmented by i.c.v. NPY to 8.7 +/- 0.9 g and 14.4 +/- 1.5 g, respectively (P less than 0.05). This increase in food consumption was due to a prolongation of feeding time. The opioid receptor antagonist naloxone significantly augmented latency to feed, both in the absence and presence of NPY (8.0 vs 1.7 min or 14.7 vs 2.8 min, respectively) and abolished the NPY-induced increase in food intake. Following intrahypothalamic injection of NPY, an increase in food intake (greater than 20%) was observed in 50% of the histologically identified LH and VMH sites, but only in 15% of the injection sites outside the LH/VMH.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sodium consumption following lesions surrounding the anteroventral third ventricle

Consumption of sodium solution was measured in rats with lesions of the anteroventral third ventricle (AV3V) and maintained on sodium deficient chow. Rats with AV3V lesions consumed less sodium than control animals, and increased their sodium consumption following a subcutaneous injection of Formalin. These data indicate that the AV3V ablation alters the nature of the regulatory control of sodium but does not render animals insensitive to plasma sodium levels.     

Neuronal and neurovascular integration following transplantation of the fetal hypothalamus into the third cerebral ventricle of adult Brattleboro rats. Neurological transplants: I

This investigation has confronted some very basic questions of neurobiology and specifically deals with the neurovascular and neuroanatomical interactions that occur between graft and host following neural transplantation. Host Long-Evans rats with chronic autosomal diabetes insipidus (DI) received stereotaxic implants of normal 17 day post-coitus fetal hypothalamic fragments from the rostral (anterior) hypothalamus of normal Long-Evans pups. Following stereotaxic surgery DI hosts were killed 60 or 90 days later and their brain prepared for correlative microangiography-immunocytochemistry coupled with transmission electron microscopy. Explants were rapidly invaded by host vessels from several routes. (1) Vessels appeared to arise from portal capillaries in the underlying median eminence and (2) from adjacent vessels from the paraventricular nucleus and surrounding endocrine hypothalamus and (3) possibly from intrinsic vessels of the graft. The former remained fenestrated and established bonafide neurovascular zones in the ventral regions and in actively growing explants. Small clusters of arginine vasopressin-positive fibers and neurophysin positive neurons were noted throughout the parenchyma of explants. Despite the presence of neurosecretory neurons and neurovascular (neurohemal) zones, none of the host rats exhibited a physiological return to normal parameters of water balance. However the active growth and development of explants in the third cerebral ventricle of DI host rats coupled with emergence of neurovascular zones lends support to a potential model for analyzing the development of anatomical substrates for the central delivery of neuropeptide hormones.     

Passive avoidance impairment in rats following cycloheximide injection into the amygdala

Cycloheximide (CHX:1, 10 or 20 microg) was injected via indwelling cannulas into various regions of the rat brain and its effects on passive avoidance training were studied. Rats with 10 or 20 microg of CHX injected into the amygdala immediately after the training footshock exhibited amnesia for the learning experience when tested after 24 h. In contrast, animals injected with 20 microg of CHX at a site either in the internal capsule only 2 mm above the amygdaloid injection site or in the frontal cortex showed no retention deficit when tested after 24 h. A quantitative examination of protein synthesis in brain halves 30 min after unilateral injection of 20 microg of CHX into the amygdala demonstrated that total protein synthesis was inhibited by less than 10%. Autoradiographic studies revealed that this inhibition resulted from a profound, highly localized inhibition of protein synthesis in areas immediately adjacent to the cannula. A comparison of the regional patterns of protein synthesis inhibition caused by injection of CHX into either the amygdala or internal capsule suggested that CHX might produce amnesia by virtue of its localized effect on the amygdala. Control experiments revealed that injection of 20 microg CHX into the amygdala had no effect on short-term retention, or short-term performance. Injection of 20 microg of CHX into the amygdala 12 h after the footshock had no effect on long-term retention. The observed impairment of retention was shown to be dose-dependent as injection of 1 microg of CHX into the amygdala was without effect. In addition, it was demonstrated that the CHX-induced amnesia did not result from induction of local seizure activity. These data show that localized injections of small amounts of CHX into the amygdala can produce deficient memory of a training experience even though total brain protein synthesis is only slightly inhibited.     

Intermittent, chronic fenfluramine administration to rats repeatedly suppresses food intake despite substantial brain serotonin reductions

The mechanisms by which fenfluramine suppresses food intake and body weight have been linked to its ability to enhance transmission across serotonin synapses in brain. This drug initially lowers body weight and suppresses food intake, yet after repeated administration food intake soon returns to normal and body weight no longer decreases. Fenfluramine also causes rapid and prolonged reductions in brain serotonin concentrations, which might account for its loss of appetite suppression. This possibility has been evaluated in rats by assessing if intermittent, chronic fenfluramine administration could suppress food intake during each treatment period, and if so, whether such an effect occurs in the presence of reduced brain serotonin levels. Rats were injected once daily with 10 mg/kg D,L-fenfluramine for 5 days, and then received no injections for the next 5 days. Control rats received only vehicle injections. This 10-day sequence was repeated five more times. During each period of fenfluramine administration, daily food intake dropped markedly the first 1-2 days of treatment, but returned to pretreatment values by day 5. Daily food intake was normal or slightly above normal during non-injection periods. Body weight dropped modestly during each period of fenfluramine administration, and rose during each subsequent period when injections had ceased. Serotonin concentrations and synthesis rates in several brain regions were markedly reduced at early, middle, and late periods of the experiment. Despite the long-term reduction in brain serotonin pools produced by fenfluramine, the drug continues to reduce food intake and body weight. Several possible interpretations of these findings are considered, based on the multiple mechanisms through which this drug has been proposed to modify synaptic serotonin transmission.     

Effects of paraventricular nucleus injection of CCK-8 on plasma CCK-8 levels in rats

The aim of the study was to determine whether paraventricular nucleus (PVN) injection of an anorexic 500-pmol dose of cholecystokinin (CCK)-8 could increase plasma CCK-8 levels sufficiently to suppress feeding by a peripheral mechanism. Rats received PVN injections of CCK-8 either alone or with ³H-labelled propionylated CCK-8 (³H-pCCK-8) and plasma samples were taken at various times from 3 to 120 min post-injection. Plasma CCK-8 levels were estimated from measurements of both total plasma CCK-like immunoreactivity (CCK-LI) and ³H-pCCK-8 activity. PVN injections of CCK-8 and ³H-pCCK-8 produced estimated peak increases in plasma CCK-8 of 15±11 and 22±3 pM, respectively. The i.v. infusion of CCK-8 doses (0.2 and 1 nmol/kg h) that bracketed the threshold dose for suppression of feeding, increased plasma CCK-LI from a basal level of 6±1 to 49±10 and 166±36 pM, respectively. The i.v. injections of 600 and 4800 pmol of CCK-8 did not suppress feeding. These results suggest that PVN injection of an anorexic 500-pmol dose of CCK-8 does not increase plasma CCK-8 levels sufficiently to suppress feeding by a peripheral mechanism.     

CART in the dorsal vagal complex: sources of immunoreactivity and effects on Fos expression and food intake

CART-peptide (CARTp) has been shown to suppress food intake, particularly when injected into the 4th ventricle of rats, and the presence of CART in nodose ganglia suggested a role in satiation. Based on retrograde tracing from the DVC combined with CART immunohistochemistry and supranodose vagotomy, we found that CART immunoreactivity in varicose fibers of the dorsal vagal complex originates from vagal afferents, sparse projections from the medullary reticular formation and the arcuate/retrochiasmatic nucleus of the hypothalamus, and most likely also from local CART neurons in the area postrema and NTS. In the nodose ganglia, 17% of neurons with projections to the stomach and 41% to the duodenum express CART-IR. CART-IR vagal afferents significantly contribute to the rich fiber plexus in mainly the commissural NTS and the adjacent area postrema. Injections of CARTp into the 4th ventricle strongly suppressed sucrose drinking and stimulated expression of c-Fos in the NTS. Injections of CARTp directly into various subnuclei of the NTS were less effective in suppressing food intake. The findings suggest that the critical site for CART's suppression of food intake is not in the termination zone of CART-containing vagal afferents in the commissural NTS, and that CART release from vagal afferent terminals plays a minor role in satiation. The functional role of CART in vagal afferents and the site of food intake suppression by 4th ventricular CARTp remain to be determined.     

C-fos protein expression in the nucleus of the solitary tract correlates with cholecystokinin dose injected and food intake in rats

C-fos protein expression was investigated in the nucleus of the solitary tract (NTS) in response to increasing cholecystokinin (CCK) doses and food intake in rats by counting the number of c-fos protein positive cells in the NTS. C-fos protein expression in the NTS dose-dependently increased in response to CCK, the lowest effective dose being 0.1 μg/kg. The ED₅₀ for c-fos protein expression in the NTS in response to CCK was calculated to be 0.5 to 1.8 μg/kg, depending on the anatomical level of the NTS investigated. Food intake increased c-fos protein expression in the NTS, the maximum number of c-fos protein positive cells being reached at 90 min after the start of food intake. Regression analysis identified a positive correlation between c-fos protein expression and the amount of food intake. Our data indicate that subpopulations of the NTS that are activated by CCK or food intake are involved into the short-term regulation of food intake and the neural control of feeding by the caudal brainstem.     

Meningioma of the third ventricle

A case of meningioma of the third ventricle is described. The tumor was completely removed using a transcallosal approach.     

Food intake increased after injection of adrenaline into the median raphe nucleus of free-feeding rats

The present study examined the effects of local injections of adrenaline (AD) or noradrenaline (NA) in equimolar doses (6, 20, and 60 nmol) into the median raphe nucleus (MRN) on ingestive and non-ingestive behaviors of free-feeding rats. The results showed that the treatment with AD at doses of 20 and 60 nmol increased food intake. While the hyperphagic response evoked by 60 nmol dose of AD was accompanied by a reduction of the latency to start feeding and an increase in the frequency of feeding, the 20 nmol dose of AD was unable to change these behavioral aspects of feeding response. The meal size and non-ingestive behaviors were not affected by AD treatment in the MRN. While water intake remained unchanged after the treatment with 20 nmol of AD in the MRN, this dose decreased the latency to start drinking. Feeding and drinking behaviors were not affected by treatment with NA in the MRN. These data suggest that adrenergic receptors of MRN participate in mechanisms that control food intake initiation or appetite. In addition, our results also indicate that the availability of energetic substrate could affect the adrenergic influence on MRN neurons since previous data indicated that the injection of AD into the MRN of food restricted rats decreased food intake.     

Central tachykinin injection potently suppresses the need-free salt intake of the female rat

Repeated sodium depletions produce a persistent, enhanced need-free salt intake in the rat, particularly in the female. The neurochemical mechanisms underlying the phenomenon are still unknown. The present studies evaluated the effect on the enhanced need-free salt intake of the female rat (1) of pharmacological interference with the natriorexigenic hormones angiotensin II and aldosterone and (2) of the central injection of the tachykinin peptides, which are endowed with antinatriorexic activity. The need-free salt intake of the female rat is not modified by treatment with the angiotensin-converting enzyme inhibitor captopril or by the aldosterone receptor antagonist RU-28318. On the other hand, the behavior is highly sensitive to the inhibitory effect of central tachykinins, suggesting the possibility that need-free salt intake might be linked to modification (down-regulation) of the inhibitory tachykininergic system.