Impact of electrical stimulation of the stomach on gastric distension-induced emesis in the musk shrew.

BACKGROUND: Gastric electrical stimulation (GES) is implicated as a potential therapy for difficult-to-treat nausea and vomiting; however, there is a lack of insight into the mechanisms responsible for these effects. This study tested the relationship between acute GES and emesis in musk shrews, an established emetic model system. METHODS: Urethane-anesthetized shrews were used to record emetic responses (monitoring intra-tracheal pressure and esophageal contractions), respiration rate, heart rate variability, blood pressure, and gastrointestinal electromyograms. We investigated the effects of acute GES pulse duration (0.3, 1, 5, and 10 ms), current amplitude (0.5, 1, and 2 mA), pulse frequency (8, 15, 30, and 60 Hz), and electrode placement (antrum, body, and fundus) on emesis induced by gastric stretch, using a balloon. KEY RESULTS: There were four outcomes: (i) GES did not modify the effects of gastric stretch-induced emesis; (ii) GES produced emesis, depending on the stimulation parameters, but was less effective than gastric stretch; (iii) other physiological changes were closely associated with emesis and could be related to a sub-threshold activation of the emetic system, including suppression of breathing and rise in blood pressure; and (iv) a control experiment showed that 8-OH-DPAT, a reported 5-HT1A receptor agonist that acts centrally as an antiemetic, blocked gastric stretch-induced emesis. CONCLUSIONS AND INFERENCES: These results do not support an antiemetic effect of acute GES on gastric distension-induced emesis within the range of conditions tested, but further evaluation should focus on a broader range of emetic stimuli and GES stimulation parameters.

Role of vagal afferent innervation in feeding and brain Fos expression produced by metabolic inhibitors.

Hepatic vagal afferent fibers have been implicated in the feeding responses initiated by administration of 2,5-anhydro-D-mannitol (2,5-AM; an inhibitor of hepatic metabolism) and methyl palmoxirate (MP; an inhibitor of fat metabolism). 2,5-AM and MP also increase brain Fos expression, an indicator of neural activity, which suggests that Fos expression can reveal the central neural pathways involved in the stimulation of feeding by these agents. To more closely test the hypothesis that brain Fos expression is related to the effects of 2,5-AM and MP on feeding, the vagus was lesioned by application of capsaicin, which destroys afferent fibers, directly to the cervical vagi. Perivagal capsaicin treatment blocked 2,5-AM-induced eating and attenuated MP-induced eating. Although perivagal capsaicin treatment attenuated MP-induced Fos expression, capsaicin treatment did not affect brain Fos expression produced by 2,5-AM. It is concluded that (1) brain Fos expression is not always related to the effects of 2,5-AM on feeding, (2) capsaicin-sensitive hepatic vagal afferent fibers carry the signal that stimulates feeding following 2,5-AM treatment, and (3) MP-induced feeding and brain Fos expression is mediated in part by capsaicin-sensitive fibers.

Further studies of bulk and orosensory decrement in producing satiation of feeding in Aplysia.

Prior evidence has suggested that meal satiation in the marine mollusk Aplysia is associated with stretch of the crop. The current data, however, suggest that under some conditions, bulk in the crop can be dissociated from the propensity to feed. The crop was hyper-distended 6 h after a satiating meal of rehydrated seaweed; that is, the crop took in water and therefore contained a greater volume than it had contained immediately after satiation. Animals presented with food 6 h after an initial satiating meal consumed a new meal despite the fact that their crop was distended beyond the level at which they had previously terminated feeding. This unexpected result led to additional experiments designed to study possible orosensory decrement during presentation of food. Orosensory input was assessed by recording from the metacerebral cell (MCC) in free-moving animals. The MCC receives excitatory input in response to chemosensory stimulation of the lips, and exhibited a slow decrement during the course of a meal or during repeated lip stimulation without ingestion. Lesions of the cerebro-buccal connectives abolished the long-term MCC response decrement to lip stimulation. This result suggests that the MCC long-term response decrement to lip stimulation is a product of buccal-ganglion feedback and may not reflect sensory decrement of chemosensory pathways. Therefore, satiation may not produce a change in lip sensitivity to chemosensory input. Our data suggest that one important factor that determines satiation is a stretch stimulus of the posterior esophagus/anterior crop. This stretch stimulus may subside over several hours as the crop contents are redistributed or as receptors slowly adapt.

Insulin prohormone processing, distribution, and relation to metabolism in Aplysia californica.

The first Aplysia californica insulin gene is characterized and its proteolytic processing from prohormone to final peptides elucidated using a combination of biochemical and mass spectrometric methods. Aplysia insulin (AI) is one of the largest insulins found, with a molecular weight of 9146 Da, and an extended A chain compared with other invertebrate and vertebrate insulins. The AI prohormone produces a series of C peptides and also a unique N-terminally acetylated D peptide. AI-producing cells are restricted to the central region of the cerebral ganglia mostly within the F and C clusters, and AI is transported to neurohemal release sites located on the upper labial and anterior tentacular nerves. The expression of AI mRNA decreases when the animal is deprived of food, and injections of AI reduce hemolymph glucose levels, suggesting that the function of insulin-regulating metabolism has been conserved.

Decrement of the response of a serotonergic modulatory neuron (the metacerebral cell) in Aplysia, during repeated presentation of appetitive (food) stimuli.

Application of food (seaweed, SW) stimuli to the lips evokes a burst of metacerebral cell (MCC) spikes, and it was found in free-moving animals that repeated presentation of the stimulus was associated with a rapid decrement of the evoked responses, even in the absence of ingestion of the food. To aid in discriminating between mechanisms that may be responsible for this decrement, SW was applied repeatedly to the lip ipsilateral or contralateral to one of the paired MCCs, and then generalization of the response decrement was tested by applying a SW stimulus to the opposite (non-stimulated) receptive field. There was statistically significant generalization of response decrement and the amount of generalization appeared to be a function of whether the decrementing stimuli were presented on the side ipsilateral vs. contralateral to the recorded MCC. The overall data suggest that MCC response decrement to repeated food stimuli results in a process analogous to behavioral habituation, and the data are consistent with a simple neural model.

Neural substrate for an integrated metabolic control of feeding behavior.

Evidence indicates that feeding behavior in rats is controlled by a mechanism that integrates information about different aspects of fuel metabolism. We investigated the neural substrate for this integrated control by measuring the effect of metabolic inhibitors given alone and in combination on food intake and neuronal activity as reflected by the expression of c-Fos protein. Combined administration of methyl palmoxirate (5 mg/kg po), an inhibitor of fatty acid oxidation, and 2,5-anhydro-D-mannitol (150 mg/kg ip), which decreases liver ATP content, increased feeding in rats more than expected on the basis of eating responses after treatment with either inhibitor given alone. Combined treatment also produced a synergistic increase in Fos-like immunoreactivity in several brain areas, including the nucleus of the solitary tract, area postrema, and parvocellular portion of the hypothalamic paraventricular nucleus. These findings provide strong evidence for the involvement of selected brain regions in the metabolic control of food intake and suggest that metabolic information used to control feeding behavior is integrated in the periphery or at the level of the brain stem.

Evidence that hemolymph glucose in Aplysia californica is regulated but does not affect feeding behavior.

Hemolymph glucose increased following a meal of a commercially available dried seaweed (laver) in Aplysia californica (Aplysia). Glucose injected into the hemocoel did not affect meal size, bite latencies, swallowing rate, or 24-hr food intake. The authors found that injection of a homogenate of nerves containing a putative Aplysia insulin-like substance decreased hemolymph glucose. The nerve homogenate, however, did not affect feeding behavior. Injection of 2-deoxy-D-glucose was found to increase hemolymph glucose, an indication of gluco-privation, but instead of increasing feeding it either had no effect or, at high doses, debilitated animals and interfered with feeding. These studies suggest that glucose may be physiologically regulated in Aplysia, but it does not appear to play a role in the control of feeding behavior.

Brain fos-like immunoreactivity in chronic decerebrate and neurologically intact rats given 2,5-anhydro-D-mannitol.

Injection of the fructose analogue, 2,5-anhydro-d-mannitol (2,5-AM), increases food intake and Fos-like immunoreactivity (Fos-li) in both brainstem and forebrain structures. Because of the interconnections between brainstem and forebrain areas, it has not been possible to determine whether or to what extent induction of Fos-li in a given region reflects brainstem-forebrain interactions. We addressed this issue using chronic decerebrate (CD) rats with complete transections of the neuroaxis at the meso-diencephalic juncture. CD and neurologically intact control rats were injected (i.p.) with saline or 400 mg/kg 2,5-AM and brains were examined for Fos-li. Both intact and CD rats showed increased Fos-li in the nucleus of the solitary tract (NTS) after injection of 2,5-AM as compared with saline. 2, 5-AM treatment increased Fos-li in the external lateral division of parabrachial nucleus (PBNel) in intact but not in CD rats, suggesting that descending projections from the forebrain may play a role in the activation of PBNel neurons after 2,5-AM injection. Decerebration eliminated significant 2,5-AM-induced Fos-li responses in forebrain structures, including the paraventricular nucleus, supraoptic nucleus, bed nucleus of the stria terminalis and central nucleus of the amygdala. The results are consistent with the hypothesis that the activation of forebrain structures after 2,5-AM treatment is due to stimulation by ascending projections from the brainstem.

Metabolic inhibition increases feeding and brain Fos-like immunoreactivity as a function of diet.

Whether administration of 2,5-anhydro-D-mannitol (2,5-AM) or methyl palmoxirate (MP) elicits eating behavior in rats depends on the composition of the maintenance diet. To assess whether specific brain sites are involved in triggering the eating responses to these metabolic inhibitors, we measured food intake and Fos-like immunoreactivity (Fos-li) in rats maintained on either a low-fat/high-carbohydrate (LF/HC) or high-fat/low-carbohydrate (HF/LC) diet. Rats fed the LF/HC diet increased food intake after administration of 2,5-AM (200 mg/kg ip) but not after treatment with MP (10 mg/kg po), whereas rats maintained on the HF/LC diet increased food intake in response to MP administration but not after 2,5-AM injection. The effects of these inhibitors on brain Fos-li in several specific brain nuclei paralleled those on feeding behavior; that is, the number of cells showing Fos-li increased only under dietary conditions in which 2,5-AM or MP stimulated eating. These results suggest that the eating response to metabolic inhibition is tied to increased neuronal activity in brain regions that process vagal afferent signals.

Methyl palmoxirate increases eating behavior and brain Fos-like immunoreactivity in rats.

Administration of methyl palmoxirate (MP), an inhibitor of fatty acid oxidation, stimulates eating behavior in rats. Fos immunohistochemistry was used to determine neural pathways that may play a role in the eating response to MP. The number of cells showing Fos-like immunoreactivity (Fos-li) was quantified by computerized image analysis. MP treatment, at a dose that increased food intake (10 mg/kg, p.o.), induced Fos expression in the nucleus of the solitary tract, area postrema, lateral parabrachial nucleus, central lateral nucleus of the amygdala, dorsal lateral bed nucleus of the stria terminalis, and the paraventricular nucleus of the hypothalamus. The results suggest that MP activates an afferent pathway projecting from the hindbrain to the forebrain, which may be involved in the eating response after MP treatment.

2,5-Anhydro-D-mannitol induces Fos-like immunoreactivity in hindbrain and forebrain: relationship to eating behavior.

Injection of the fructose analogue, 2,5-anhydro-D-mannitol (2,5-AM), stimulates eating behavior in rats. Previous studies have shown that administration of 2,5-AM in doses that elicit eating induces Fos-like immunoreactivity (Fos-li) primarily in hindbrain structures, including the nucleus of the solitary tract (NTS), area postrema (AP), and lateral parabrachial nucleus (PBN). To more closely assess the relationship between neural activation and the eating response to 2,5-AM treatment, we measured food intake and brain Fos-li in rats given a range of doses of 2,5-AM. The numbers of neurons showing Fos-li were quantified by computerized image analysis. Doses of 2,5-AM that reliably stimulated food intake induced Fos-li in both the hindbrain and forebrain, including in the NTS, AP, lateral PBN, central lateral nucleus of the amygdala, dorsal lateral bed nucleus of the stria terminalis (BNSTdl), anterior paraventricular nucleus of the thalamus, supraoptic nucleus, subfornical organ, and paraventricular hypothalamic nuclei. A low dose of 2,5-AM that did not elicit eating increased Fos-li marginally only in the AP, PBN, and BNSTdl. The results suggest that 2,5-AM treatment activates a vagal afferent pathway projecting from the hindbrain to forebrain that is involved in initiating the eating response to the fructose analogue.

Does ingested fat produce satiety?

Administration of fat directly into the gastrointestinal tract of rats produces a rapid and often substantial reduction of feeding behavior. This contrasts with the normal consumption of a fat meal, which produces little change in subsequent food intake. To determine whether procedural differences account for this discrepancy, we examined the satiating effect of ingested fat on food intake of rats maintained under feeding conditions similar to those employed in studies involving gastrointestinal delivery of fat (i.e., food deprivation, liquid diet). Ingestion of approximately 1.5 ml corn oil had no effect on subsequent liquid diet intake until 90 min after oil ingestion. When rats ingested oil 4 h before access to the liquid diet, to allow time for additional gastrointestinal clearance, liquid diet intake was reduced by 13% in the first 30 min of access. These findings indicate that ingested fat decreases short-term intake slightly, but only if time is allowed for postabsorptive delivery. The results question the physiological significance of the marked suppression of food intake observed in response to administration of fat directly into the gastrointestinal tract.

Does selective vagotomy affect conditioned flavor-nutrient preferences in rats?

We investigated the effects of selective vagotomy on the learning of conditioned flavor preferences associated with intragastric Polycose infusion. Normal control, hepatic branch vagotomized, and gastric vagotomized rats implanted with gastric catheters received a flavor (CS+) paired with intragastric Polycose infusion, and on alternate days, a different flavor (CS-) paired with intragastric water infusion. After training, rats were given a two-bottle extinction test for 12 days. The results show that normal control and hepatic branch vagotomized rats had a significant CS+ flavor preference during extinction testing, whereas the gastric vagotomized rats showed no preference. This result indicates that the hepatic branch of the vagus does not play a solitary role in learned flavor-nutrient preferences using Polycose. However, the gastric vagal branches may be involved in the preference learning.

Paired associate learning in reading-disabled children: evidence for a rule-learning deficiency.

Two experiments examined whether normal and disabled readers differed in the utilization of rules in a paired associate learning task. Experiment 1 required children to learn symbol-word associates. Children were assigned to one of three conditions: nonrule, consistent rule, or inconsistent rule. When present, the rule was based on semantic opposites. Subjects benefited from having the rule, but disabled readers showed less improvement across four test blocks than both chronological age (CA) controls and reading age (RA) controls, particularly in the inconsistent condition. Experiment 2 required subjects to learn symbol-symbol associations in one of three conditions: nonrule, consistent rule, or inconsistent rule. When present, the rule specified the locations of a subsidiary figure in each symbol according to the pattern top-right, bottom-left. Disabled and normal readers did not differ in the nonrule condition where reliance on visual memory would be an effective strategy. Normal readers were superior to disabled readers in both rule conditions. In addition, disabled readers in the inconsistent rule condition were less able than normal readers to apply the rule in a generalization task where memory demands were reduced. Results supported the hypothesis that disabled readers have greater difficulty than normal readers inducing and/or using rules, particularly when they are inconsistent. It is suggested that difficulties in acquiring or using complex and inconsistent rules may be one important source of problems learning spelling-sound correspondence rules, which in English are complex and inconsistent.

Abnormal lymphocyte response to u.v. radiation in multiple skin cancer.

The lymphocyte response to u.v. radiation (254 nm) was investigated by two different methods in 29 unselected patients with multiple epidermal cancer. The u.v.-induced DNA synthesis was determined as the increase in incorporation of [3H]thymidine in irradiated cells compared with nonirradiated cells after incubation for 2 h. The u.v. tolerance was measured as the u.v. dose necessary for 50% reduction in phytohemagglutinin-stimulated lymphocyte proliferation. Patients with both squamous cell differentiated tumors and basal cell carcinomas had very high u.v.-induced DNA synthesis values (p less than 0.01, when compared with patients with basal cell carcinoma only and p less than 0.005, when compared with controls). The u.v. tolerance in patient lymphocytes was considerably lower than in control lymphocytes (p less than 0.001), with the lowest values occurring in patients with clinical sun intolerance (p = 0.05, when compared with the remaining patients). These investigations may be of predictive value in skin carcinogenesis.