Abdominal Surgery- and Trephination-Induced Delay in Gastric Emptying is Prevented by Intracisternal Injection of CRF Antagonist in the Rat

The role of endogenous corticotropin-releasing factor (CRF) in mediating surgery-induced delay in gastric emptying of a noncaloric solution was studied in rats using intracisternal injection of a CRF antagonist, α-helical CRF9-41. Intracisternal injections and surgery (abdominal or trephination) were performed under ether anesthesia. After rats recovered from anesthesia, a methylcellulose phenol red solution was administered oropharyngeally and gastric emptying was measured 20 minutes later. Laparotomy, followed by exteriorization of the cecum for 60 seconds (abdominal surgery), trephination, intracisternal injection, of CRF (130 pmol), and ether exposure alone for 120 to 140 or 70 to 80 seconds, inhibited gastric emptying by 83%, 81%, 60%, 56%, and 32%, respectively, as compared with an untreated control group. Intracisternal injection of α-helical CRF9-41 significantly enhanced gastric emptying by 27% and prevented by 79% to 87% the inhibition of gastric emptying induced by intracisternal injection of CRF and ether exposure (70–80 s) alone or combined with abdominal surgery. Intracisternal injection of the CRF antagonist (13–26 nmol) dose dependently prevented ether-induced (120–140 s) and trephination-induced inhibition of gastric emptying. These results suggest that central CRF may be involved in mediating the acute postoperative delay in gastric emptying.     

Raphe pallidus stimulation increases gastric contractility via TRH projections to the dorsal vagal complex in rats

The role of thyrotropin releasing hormone (TRH) in the dorsal vagal complex (DVC) in mediating the enhanced gastric contractility induced by glutamate (100 pmol) microinjected into the raphe pallidus (Rpa) was investigated in urethane-anesthetized rats acutely implanted with miniature strain gauge force transducers on the corpus of the stomach. Glutamate-induced stimulation of gastric contractility was dose-dependently inhibited by bilateral microinjection into the DVC of TRH antibody (0.17, 0.85 or 1.7 μg/100 nl/site) but not by vehicle. TRH antibody microinjected into the dorsal medullary reticular field had no effect. These data indicate that activation of Rpa neurons by glutamate increases gastric motor function through TRH release in the DVC.     

A magnetic device to stimulate selected whiskers of freely moving or restrained small rodents: its application in a deoxyglucose study

After receiving an intraperitoneal injection of [¹⁴C]2-deoxy-d-glucose (2-DG), a total of 28 mice which had pieces of metal wire glued to certain whiskers (all others were clipped) were exposed to magnetic field bursts. The stimulated whiskers were B1 (freely moving mice, set I) or whiskers C1–3 and E1 (restrained mice, set II) on the left side. In set I, stimulated mice were compared with animals of various control groups. Autoradiography demonstrated an activation of columnar shape overlying the presumed corresponding barrel contralateral to stimulation; in a part of the ipsilateral barrelfield, 2-DG uptake was depressed significantly. In the subnuclei caudalis and interpolaris of the trigeminal brainstem complex a spot of activation was observed ipsilaterally but there was no depression contralaterally. Whereas several animals of the control groups showed some aspects of these responses, they were consistent only in stimulated mice. In set II, animals received stimulation with different intensities. 2-DG uptake was higher in barrels C1–3 than in E1. It increased with increasing intensity. The same observations were made in two nuclei of termination. The device we describe here can be used to study stimulus-specific responses at various levels of the somatosensory pathway.     

Intracisternal PYY inhibits gastric lesions induced by ethanol in rats: role of PYY-preferring receptors?

We previously reported that intracisternal (i.c.) injection of peptide YY (PYY) and low doses of thyrotropin-releasing hormone (TRH) or TRH analog, RX 77368, increased the resistance of the gastric mucosa to ethanol injury through vagal pathways in rats. The gastroprotective effect of i.c. injection of PYY/neuropeptide NPY (NPY) agonists with differential in vitro affinity to the Y receptor subtypes was examined in urethane-anesthetized rats. Intragastric administration of ethanol (45%, 5 ml/kg) results in mucosal lesions covering 23+/-2% of the gastric corpus in 1 h. PYY (500 ng, i.c.) significantly reduced ethanol-induced gastric lesions by 52%. [Pro34]PYY (PYY-preferring/Y1/Y5/Y4 subtypes) injected i.c. at 50, 100, 200 or 500 ng, reduced dose dependently gastric lesions to 15.4+/-2.2%, 11.4+/-3.1%, 8.6+/-2.9% and 5.4+/-2.2%, respectively. PYY3-36, (Y2/Y4 subtypes), [Leu31, Pro34]NPY (Y1/Y5), NPY (Y3/Y1/Y5/Y2) and pancreatic polypeptide (PP, Y4) injected i.c. at 500 ng did not influence significantly ethanol-induced gastric lesions. Combined i.c. injection of RX 77368 (1 ng) and Pro34PYY (25 ng), at sub-threshold doses given singly, reduced ethanol-induced gastric injury to 12.9+/-2.3% while RX 77368 (1 ng) plus PYY3-36 (500 ng) or [Leu31, Pro34]NPY (25 ng) had no effect. These findings indicate that i.c. PYY-induced gastric protection against 45% ethanol is mediated by a Y receptor subtype which bears similarity with the putative PYY-preferring receptor and distinct from the currently defined Y1/Y5; in addition, there is a synergistic interaction between activation of this PYY-preferring receptor and i.c. TRH to increase the resistance of the gastric mucosa to injury caused by 45% ethanol.     

Role of brainstem TRH/TRH-R1 receptors in the vagal gastric cholinergic response to various stimuli including sham-feeding

Pavlov's pioneering work established that sham-feeding induced by sight or smell of food or feeding in dogs with permanent esophagostomy stimulates gastric acid secretion through vagal pathways. Brain circuitries and transmitters involved in the central vagal regulation of gastric function have recently been unraveled. Neurons in the dorsal vagal complex including the dorsal motor nucleus of the vagus (DMN) express thyrotropin-releasing hormone (TRH) receptor and are innervated by TRH fibers originating from TRH synthesizing neurons in the raphe pallidus, raphe obscurus and the parapyramidal regions. TRH injected into the DMN or cisterna magna increases the firing of DMN neurons and gastric vagal efferent discharge, activates cholinergic neurons in gastric submucosal and myenteric plexuses, and induces a vagal-dependent, atropine-sensitive stimulation of gastric secretory (acid, pepsin) and motor functions. TRH antibody or TRH-R1 receptor oligodeoxynucleotide antisense pretreatment in the cisterna magna or DMN abolished vagal-dependent gastric secretory and motor responses to sham-feeding, 2-deoxy-D-glucose, cold exposure and chemical activation of cell bodies in medullary raphe nuclei. TRH excitatory action in the DMN is potentiated by co-released prepro-TRH-(160-169) flanking peptide, Ps4 and 5-HT, and inhibited by a number of peptides involved in the stress/immune response and inhibition of food-intake. These neuroanatomical, electrophysiological and neuropharmacological data are consistent with a physiological role of brainstem TRH in the central vagal stimulation of gastric myenteric cholinergic neurons in response to several vagal dependent stimuli including sham-feeding.     

Increased local uptake of 2-deoxyglucose after electrochemical or direct deposition of iron into the rat brain

Intracerebral application of Fe^{2+, 3+} has been widely used to study mechanisms controlling the release of anterior pituitary hormones. Yet its local effects on neuronal function remain controversial. The present study utilized radiolabeled 2-deoxy-d-glucose (2-DG) incorporation to assess acute neural metabolic effects at the site of iron deposition. Fe^{2+, 3+} was applied either directly or electrochemically to preoptic and hypothalamic regions. Application of Fe^{2+, 3+} resulted in a local increase of 2-DG uptake that succeeded the more transient effect of NaCl injection or anodic direct current applied through platinum electrodes which served as controls. In several cases, a clear pattern of multilaminar rings was evident in autoradiograms. These results are considered as providing additional evidence that the short-term effect of Fe^{2+, 3+} is to increase neural activity.     

Bombesin microinjected into the dorsal vagal complex inhibits TRH-stimulated gastric contractility in rats

The effects of centrally injected bombesin on central and peripheral stimulated gastric contractility were investigated in fasted urethane-anesthetized rats. Miniature strain gauge force transducers were acutely implanted on the corpus of the stomach and gastric contractility was analyzed by computer. Intracisternal injection of the stable thyrotropin-releasing hormone (TRH)-analog RX 77368 (77 pmol) induced a stimulation of gastric contractility for 40 min. Intracisternal injection of bombesin (62-620 pmol) followed 30 min later by that of RX 77368 resulted in a dose-related inhibition of the TRH-analog-induced gastric contractility. Intracisternal injection of bombesin (620 pmol) did not modify gastric contractility stimulated by intravenous carbachol. Stimulation of gastric contractility induced by TRH-analog microinjected into the dorsal vagal complex (DVC) was dose-related suppressed by concomitant injections of bombesin (6.2-620 pmol). Neither bombesin alone (6.2 pmol) nor vehicle modified basal gastric contractility. These results demonstrate that bombesin acts within the brain to inhibit vagally stimulated gastric contractility and that the DVC is a sensitive site for bombesin inhibitory action. These findings suggest a possible interaction between TRH and bombesin in the central vagal regulation of gastric contractility.     

2-deoxy-D-glucose modulation of T-lymphocyte reactivity: differential effects on lymphoid compartments

This study was designed to evaluate the effect of glucoprivation, as induced by 2-deoxy-D-glucose (2-DG) administration, on lymphocyte mitogen reactivity in Sprague-Dawley rats. The results showed that a single injection of 2-DG decreased reactivity in both whole-blood and spleen lymphocytes, as determined by mitogenic stimulation to concanavalin A (Con A) and phytohemagglutinin (PHA). However, the suppressed reactivity for the spleen lymphocytes attenuated with repeated injections, but the whole-blood lymphocytes did not show attenuation. Mitogen assessments of lymphocytes obtained from the thymus indicated that a single injection did not induce suppressed reactivity, but repeated injections induced a pronounced suppression of responsiveness. Furthermore, mitogen assessments of mesentery lymph nodes did not show any effect of 2-DG injections. These results corroborate other findings using electric shock as the stressor, namely that different compartments of the immune system are differentially affected by a stressor.     

TRH stimulation and L-glutamic acid inhibition of proximal gastric motor activity in the rat dorsal vagal complex

The importance of the dorsal vagal complex (DVC) in the control of gastric motor activity has been previously established by electrical and chemical stimulation of this region. We have further evaluated excitatory and inhibitory influences on motor activity of the gastric corpus by microinjection of L-glutamic acid (GLU) and thyrotropin-releasing hormone (TRH) into the DVC. GLU and TRH were ejected by pressure (20-30 psi) in 1-10 nl vol. from multibarreled micropipettes and intraluminal pressure in the gastric corpus was measured using a manometric catheter placed into the stomach through the pylorus of urethane-chloralose anesthetized rats. Gastric motor activity was monitored while micropipettes were advanced from the surface of the dorsomedial medulla to a depth of 1 mm in 100 micron increments. Microinjections of GLU (1-10 pmol) at depths of 200-600 microns below the surface of the brainstem caused a decrease in tonic intraluminal pressure and amplitude of phasic contractions of the gastric corpus. Injection of TRH (1-10 pmol) at depths of 200-800 microns increased both tonic intraluminal pressure and amplitude of phasic contractions. The responses to GLU (10 pmol) and TRH (10 pmol) were abolished by hexamethonium and vagotomy; atropine abolished the effect of TRH and attenuated that of GLU. It is concluded that GLU evokes only vagally mediated inhibitory effects on tonic and phasic gastric motor activity when microinjected into the DVC. In contrast, injection of TRH at the same loci causes only vagal cholinergic increases in motor activity. Subpopulations of neurons in the DVC may, therefore, be activated by specific neurotransmitters having opposite effects on gastric motor activity.     

Long experimental durations are required for double label [C]- and [H]2-deoxyglucose autoradiographic methods

Double-label 2-deoxyglucose (2-DG) studies using sequential [¹⁴C]- and [³H]2-DG injections demonstrate increased [¹⁴C]2-DG uptake durating the first and second stimulation periods. To understand why this occurs, the rat mystacial vibrissae were stimulated at various times following [¹⁴C]2-DG injection. Local cerebral glucose utilization (LCGU) increased when whisker stimulation was performed at 0–90 min following [¹⁴C]2-DG injection. LCGU did not increase when whisker stimulation was performed at 90–150 min following [¹⁴C]2-DG injection. To minimize contamination of the two tracers in double label 2-DG mapping studies, the time between [¹⁴C]- and [³H]2-DG administration should be increased to 90 min.     

Characterization of the Effect of 2-Deoxy-d-Glucose(2-DG) on the Immune System

This study was designed to characterize the effects of the metabolic stress of administration of 2-deoxy-d-glucose (2-DG, 500 mg/kg) on immune function. Male Lewis rats were exposed to one or five injections (one every 48 h) of 2-DG. Control rats received saline injections. Administration of 2-DG induced a reduction of total leukocytes in the spleen, thymus, and blood. The reduction was most prominent in animals that received five injections of 2-DG. The ratio of CD4⁺/CD8⁺in the spleen was decreased due to a significant increase of CD8⁺T-cell subpopulation. Additionally, 2-DG induced a suppression of mitogenic responsiveness and IFN-γ production in both whole blood and spleen lymphocytes. The production of IL-1 and IL-2 was significantly reduced in the blood, but not in the spleen. Conversely, there was a significant increase in nitric oxide production in cultures of Con A-, PHA-, and LPS-stimulated splenocytes from 2-DG-injected animals compared with saline-injected controls. In blood cultures stimulated with Con A and PHA, the nitric oxide production of the group that received five injections of 2-DG was significantly higher than in the group that received one injection of 2-DG or saline. These results demonstrated that the metabolic stress 2-DG induced a downregulation of Th 1 cellular immune function in a manner similar to physical and psychological stressors. Additionally, the use of 2-DG in rats provided an important model with which to study metabolic stress.     

Effects of vasopressin and long pulse–low frequency gastric electrical stimulation on gastric emptying, gastric and intestinal myoelectrical activity and symptoms in dogs

The aim of this study was to investigate the effect of vasopressin and long pulse-low frequency gastric electrical stimulation (GES) on gastric emptying, gastric and intestinal myoelectrical activity and symptoms in dogs. The study was performed in eight healthy female dogs implanted with four pairs of gastric serosal electrodes and two pairs of small bowel serosal electrodes, and a duodenal fistula for the assessment of gastric emptying. Each dog was studied in three sessions on three separate days in a randomized order with recordings of gastric and small bowel slow waves. Each study session consisted of 30-min baseline, 30-min stimulation and 30-min recovery period. In sessions 1 and 2, infusion of either saline or vasopressin (0.75 U kg(-1) in 30 mL saline instilled in 30 min) was given during the second 30-min period. The protocol of session 3 was the same as session 2 except long pulse-low frequency GES was performed during the second 30-min period. It was found that: (i) Vasopressin significantly delayed gastric emptying 30 and 45 min after meal and GES did not improve the vasopressin induced delayed gastric emptying; (ii) Vasopressin induced gastric dysrhythmias and GES significantly improved vasopressin induced gastric dysrhythmia; (iii) Vasopressin also induced intestinal slow wave abnormalities but GES had no effect on vasopressin induced small bowel dysrhythmia; (iv) Vasopressin induced symptoms and behaviours suggestive of nausea that were not improved by GES. We conclude that: (i) Vasopressin delays gastric emptying and induces gastric and small bowel dysrhythmias and symptoms in the fed state, and (ii) long pulse-low frequency GES normalizes vasopressin induced gastric dysrhythmia with no improvement in gastric emptying or symptoms.     

Neural mechanisms involved in the delay of gastric emptying of liquid elicited by acute blood volume expansion in awake rats

We have previously reported that acute blood volume expansion in awake rats delays the gastric emptying of a liquid meal, using the phenol red method. In this study we attempted to investigate the neural mechanisms involved in this phenomenon. Blood volume expansion, due to Ringer-bicarbonate infusion up to a volume equivalent to 5% of body weight, decreased the gastric emptying of a liquid meal by half (38.2 +/- 1.8 vs 18.7 +/- 3.2%, P < 0.05). The blood volume expansion effect on gastric emptying of liquid was prevented by separate pretreatments, consisting of subdiaphragmatic vagotomy or i.v. injection of hexamethonium (20 mg kg-1) or yohimbine (3 mg kg-1). Intravenous injection of atropine (0.5 mg kg-1), guanethidine (10 mg kg-1), L-NAME (3 mg kg-1), prazosin (1 mg kg-1) or propranolol (2 mg kg-1) did not prevent the blood volume expansion effect on gastric emptying. Bilateral adrenalectomy or coeliac ganglionectomy were also ineffective. The results indicate that blood volume expansion decreases gastric emptying of liquid through vagal-dependent pathways, sensitive to hexamethonium and yohimbine. Evidence for the participation of the peripheral sympathetic nervous system was not found.     

Role of the autonomic nervous system in the cytoprotective effect of neurotensin against gastric stress ulcers in rats

Pharmacologic agents were used to study the role of the autonomic nervous system in the cytoprotection produced by intracisternal neurotensin against cold plus restraint stress-induced gastric ulcers in rats. Drugs which stimulated alpha- or beta-adrenergic receptors or blocked muscarinic cholinergic receptors reduced the incidence of ulcers to a similar degree as intracisternal neurotensin; alpha-adrenergic or beta-adrenergic blockade as well as cholinergic stimulation prevented neurotensin's beneficial effect. However, pretreatment with indomethacin blocked only the cytoprotective effect of neurotensin or beta-adrenergic stimulation, but not that of muscarinic cholinergic blockade. In addition, pretreatment with reserpine or guanethidine also was effective in preventing cytoprotection by intracisternal neurotensin. These data indicate that the mechanism for cytoprotection by centrally administered neurotensin is mediated at least in part through activation of the sympathetic nervous system. This activation by neurotensin appears to produce cytoprotection by stimulation of gastric mucosal prostaglandin synthesis.     

Effects of gastric electrical field stimulation with long pulses on gastric emptying in dogs

The aim was to investigate the effects of electrical field stimulation (EFS) with long and short pulses on gastric emptying, gastric contractility and vagal activity in dogs. Sixteen dogs were equipped with a duodenal cannula, electrodes and strain gauges (10 dogs) in the stomach. Each dog was fed with Ensure and gastric effluent was collected from the cannula. Electrical stimulation was applied via two electrodes (about 12 cm apart, one in the corpus and the other in the antrum) with long pulses (a frequency of 6 cycles min-1, pulse amplitude of 6 mA and width of 100 ms) in 10 dogs and with short pulses (frequency of 30 Hz and pulse width of 300 micros) in six dogs. The electrocardiogram was also recorded and heart rate variability was derived to assess the vagal activity. It was found that: (i). EFS with long pulses did not alter gastric emptying during stimulation but increased gastric emptying during the 45 min immediately after stimulation; (ii). EFS with long pulses increased gastric contractility in both proximal and distal antrum during and after the stimulation; (iii). EFS with long pulses resulted in an increase in vagal tone during the 45 min immediately after stimulation. However, there is no difference during the 45 min period of stimulation; (iv). EFS with short pulses had no effect on gastric emptying. We concluded that long pulse gastric electrical field stimulation with one electrode in the corpus and the other electrode in the antrum has postponed effects on gastric emptying of liquid, gastric contractility and vagal activity.     

TRH stimulation andl-glutamic acid inhibition of proximal gastric motor activity in the rat dorsal vagal complex

The importance of the dorsal vagal complex (DVC) in the control of gastric motor activity has been previously established by electrical and chemical stimulation of this region. We have further evaluated excitatory and inhibitory influences on motor activity of the gastric corpus by microinjection ofl-glutamic acid (GLU) and thyrotropin-releasing hormone (TRH) into the DVC. GLU and TRH were ejected by pressure (20–30 psi) in 1–10 nl vol. from multibarreled micropipettes and intraluminal pressure and the gastric corpus was measured using a manometric catheter placed into the stomach through the pylorus of urethane-chloralose anesthetized rats. Gastric motor activity was monitored while micropipettes were advanced from the surface of the dorsomedial medulla to a depth of 1 mm in 100 μm increments. Microinjections of GLU (1–10 pmol) at depths of 200–600 μm below the surface of the brainstem caused a decrease in tonic intraluminal pressure and amplitude of phasic contactions of the gastric corpus. Injection of TRH (1–10 pmol) at depths of 200–800 μm increased both tonic intraluminal pressure and amplitude of phasic contractions. The responses to GLU (10 pmol) and TRH (10 pmol) were abolished by hexamethonium and vagotomy; atropine abolished the effect of TRH and attenuated that of GLU. It is concluded that GLU evokes only vagally mediated inhibitory effects on tonic and phasic gastric motor activity when microinjected into the DVC. In contrast, injection of TRH at the same loci causes only vagal cholinergic increases in motor activity. Subpopulations of neurons in the DVC may, therefore, be activated by specific neurotransmitters having opposite effects on gastric motor activity.     

9The TANTALUSTM System for obesity: effect on gastric emptying of solids

Background: Gastric electrical stimulation (GES) is currently investigated for the treatment of obesity. The TANTALUS System delivers gastric contractility modulation (GCM) signals in synchrony with gastric slow waves, resulting in significant augmentation of gastric contractions during food intake. We hypothesized that such modulation of contractile activity may affect gastric emptying and plasma ghrelin levels. Aim: To test the effect of GCM of the gastric antrum on gastric emptying of solids and ghrelin levels. Methods: 12 obese subjects were implanted with 2 pairs of antral electrodes and an implantable pulse generator (IPG, TANTALUS ^TM) Gastric emptying test (GE) for solids was performed twice, on separate days, in each subject, starting few weeks after implantation: 1) control, before the start of stimulation, and 2) with stimulation, after device was turned on. Blood samples for ghrelin, were taken at baseline, and at 15, 30, 60 and 120 min after the test meal. Results as mean + SD, analysis by t‐test and p < 0.05. Results: 11 females, 1 male, age: 39.1 ± 8.9 years, BMI: 41.6 ± 3.4, 3 subjects with type 2 diabetes. One diabetic patient did not complete GE test because of technical issues. GCM significantly accelerated gastric emptying: retention at 2 hours 18.7 ± 12.2% vs. 31.9 ± 16.4%, stimulation vs. control respectively, p = 0.008. T ^1/2 78.3 ± 23.5 vs. 95 ± 31.7 min, stimulation vs. control respectively, p = 0.04. Mean results for gastric emptying were within normal at both baseline and stimulation. Meal ingestion induced only minimal, insignificant reduction in ghrelin levels. There was no significant difference in AUC of ghrelin between control and stimulation. Conclusions: After GCM stimulation, there is significant acceleration of gastric emptying of solids in obese patients, without affect on ghrelin levels. The obese subjects did not exhibit the significant, meal‐induced reduction in ghrelin.      

Functional activation in the auditory system of the rat produced by arousing reticular stimulation: a 2-deoxyglucose study

The 2-deoxyglucose (2-DG) autoradiographic method was used to map the activity in the auditory pathway during behaviorally arousing electrical stimulation of the mesencephalic reticular formation (RET). Uptake of 2-DG during RET stimulation was compared to the effect of a frequency-modulated tone (4-5 kHz, 60 dB SPL) and to controls without stimulation. The major finding was a specific pattern of increased metabolic activation throughout the auditory pathway evoked during RET stimulation. The observed increases in 2-DG uptake were always greater in RET-stimulated rats as compared to sound-stimulated or control rats. The dorsal cochlear nucleus (DCN) showed the largest incorporation of 2-DG among the auditory nuclei of the brainstem in RET-stimulated rats. In the central nucleus of the inferior colliculus a layered pattern made of 3 discrete bands of high 2-DG uptake was visible in RET-stimulated rats. The medial geniculate (MG) and the auditory cortex (AC) also showed highly significant increases in 2-DG uptake induced by RET stimulation. The method provided correlations between classical morphological schemes of parcellation on nuclei and functionally defined areas of increased 2-DG uptake. Our observations represent the first anatomical demonstration of the activating effects of RET stimulation in a sensory system, and they support the concepts of arousing reticular mechanisms for sensory control.     

A role for central postsynapticα2-adrenoceptors in glucoregulation

Central or peripheral administration of theα₂-adrenoceptor agonist clonidine causes marked hyperglycemia in the rat. It is not clear whether this effect is mediated within the brain at either pre- or postsynapticα₂-adrenoceptors or whether it is due to peripheralα₂-agonist actions. We employed computerized mass spectrometry to measure noradrenaline (NA) and its primary neuronal metabolite 3,4-dihydroxyphenylglycol (DHPG) in the medial basal hypothalamus of rats treated acutely with clonidine, theα₂-antagonist yohimbine, the postganglionic noradrenergic blocker guanethidine and the neuroglycopenic agent 2-deoxy-d-glucose (2-DG). That clonidine's hyperglycemic effect was due, in part, to an action at centralα₂-adrenoceptors was indicated by the ability of guanethidine to significantly inhibit the glucose response. Because of clonidine's inhibition of hypothalamic NA release (assessed by the DHPG/NA ratio), presumably by presynaptic agonism, these data indicated that postsynaptic receptor stimulation by clonidine was involved in activating glucose release. Yohimbine markedly increased the hypothalamic DHPG/NA ratio, reflecting presynaptic stimulation of NA release, but at the same time inhibited the hyperglycemic response due to 2-DG administration. This latter effect to block hyperglycemia is consistent with antagonism of postsynapticα₂-adrenoceptors involved in mediating hepatic glucose output. These data indicate a major role for postsynapticα₂-adrenoceptors in glucoregulation.     

Hypothalamic galanin: control by signals of fat metabolism

The peptide, galanin (GAL), is known to stimulate eating behavior, reduce energy expenditure and affect the release of metabolic hormones. Further, the activity of this peptide in the hypothalamus is modulated, in turn, by these hormones as well as by the ingestion of nutrients. The focus of this investigation is on signals related to nutrient metabolism that may also affect GAL production and, through these neurochemical events, control the ingestion of specific nutrients. Three experiments were performed in normal-weight male, Sprague–Dawley rats. In Experiment 1, the impact of food deprivation (24 and 48 h) was examined. Experiment 2 tested the effects of the compound, 2-deoxy-d-glucose (2-DG, 200 and 400 mg/kg), which blocks glucose utilization, whereas Experiment 3 studied mercaptoacetate (MA, 200 and 600 μmol/kg), which blocks fatty acid oxidation. Eating behavior was examined in some rats, whereas hypothalamic GAL activity was measured in others using radioimmunoassay, immunohistochemistry and in situ hybridization. Both food deprivation and MA (600 μmol/kg), but not 2-DG, affected GAL in the hypothalamus, in one specific area. This is the anterior parvocellular region of the paraventricular nucleus (aPVN), which has a dense concentration of GAL-containing neurons and terminals. GAL gene expression and peptide immunoreactivity in this area is enhanced by food deprivation; in contrast, it is reduced by injection of MA. Other hypothalamic sites with dense concentrations of GAL-containing neurons or fibers are unaffected by food deprivation or MA, and the antimetabolite 2-DG has no impact on GAL in any area. Behavioral measurements indicate that these shifts in GAL activity are accompanied by specific changes in eating behavior. Food deprivation which enhances aPVN GAL produces a marked increase in fat ingestion, whereas MA which reduces aPVN GAL causes a specific reduction in fat ingestion along with a stimulation of protein intake. In contrast, 2-DG preferentially enhances ingestion of carbohydrate. These findings suggest a possible relationship between GAL activity in the aPVN and the metabolic and behavioral processes of fat metabolism and ingestion.