BACE inhibitor treatment of mice induces hyperactivity in a Seizure-related gene 6 family dependent manner without altering learning and memory.

BACE inhibitors, which decrease BACE1 (ß-secretase 1) cleavage of the amyloid precursor protein, are a potential treatment for Alzheimer's disease. Clinical trials using BACE inhibitors have reported a lack of positive effect on patient symptoms and, in some cases, have led to increased adverse events, cognitive worsening and hippocampal atrophy. A potential drawback of this strategy is the effect of BACE inhibition on other BACE1 substrates such as Seizure-related gene 6 (Sez6) family proteins which are known to have a role in neuronal function. Mice were treated with an in-diet BACE inhibitor for 4-8 weeks to achieve a clinically-relevant level of amyloid-ß40 reduction in the brain. Mice underwent behavioural testing and postmortem analysis of dendritic spine number and morphology with Golgi-Cox staining. Sez6 family triple knockout mice were tested alongside wild-type mice to identify whether any effects of the treatment were due to altered cleavage of Sez6 family proteins. Wild-type mice treated with BACE inhibitor displayed hyperactivity on the elevated open field, as indicated by greater distance travelled, but this effect was not observed in treated Sez6 triple knockout mice. BACE inhibitor treatment did not lead to significant changes in spatial or fear learning, reference memory, cognitive flexibility or anxiety in mice as assessed by the Morris water maze, context fear conditioning, or light-dark box tests. Chronic BACE inhibitor treatment reduced the density of mushroom-type spines in the somatosensory cortex, regardless of genotype, but did not affect steady-state dendritic spine density or morphology in the CA1 region of the hippocampus. Chronic BACE inhibition for 1-2 months in mice led to increased locomotor output but did not alter memory or cognitive flexibility. While the mechanism underlying the treatment-induced hyperactivity is unknown, the absence of this response in Sez6 triple knockout mice indicates that blocking ectodomain shedding of Sez6 family proteins is a contributing factor. In contrast, the decrease in mature spine density in cortical neurons was not attributable to lack of shed Sez6 family protein ectodomains. Therefore, other BACE1 substrates are implicated in this effect and, potentially, in the cognitive decline in longer-term chronically treated patients.

N-aryl piperazine metabotropic glutamate receptor 5 positive allosteric modulators possess efficacy in preclinical models of NMDA hypofunction and cognitive enhancement.

Impaired transmission through glutamatergic circuits has been postulated to play a role in the underlying pathophysiology of schizophrenia. Furthermore, inhibition of the N-methyl-d-aspartate (NMDA) subtype of ionotropic glutamate receptors (NMDAR) induces a syndrome that recapitulates many of the symptoms observed in patients with schizophrenia. Selective activation of metabotropic glutamate receptor subtype 5 (mGlu5) may provide a novel therapeutic approach for treatment of symptoms associated with schizophrenia through facilitation of transmission through central glutamatergic circuits. Here, we describe the characterization of two novel N-aryl piperazine mGlu5 positive allosteric modulators (PAMs): 2-(4-(2-(benzyloxy)acetyl)piperazin-1-yl)benzonitrile (VU0364289) and 1-(4-(2,4-difluorophenyl)piperazin-1-yl)-2-((4-fluorobenzyl)oxy)ethanone (DPFE). VU0364289 and DPFE induced robust leftward shifts in the glutamate concentration-response curves for Ca(2+) mobilization and extracellular signal-regulated kinases 1 and 2 phosphorylation. Both PAMs displayed micromolar affinity for the common mGlu5 allosteric binding site and high selectivity for mGlu5. VU0364289 and DPFE possessed suitable pharmacokinetic properties for dosing in vivo and produced robust dose-related effects in reversing amphetamine-induced hyperlocomotion, a preclinical model predictive of antipsychotic-like activity. In addition, DPFE enhanced acquisition of contextual fear conditioning in rats and reversed behavioral deficits in a mouse model of NMDAR hypofunction. In contrast, DPFE had no effect on reversing apomorphine-induced disruptions of prepulse inhibition of the acoustic startle reflex. These mGlu5 PAMs also increased monoamine levels in the prefrontal cortex, enhanced performance in a hippocampal-mediated memory task, and elicited changes in electroencephalogram dynamics commensurate with procognitive effects. Collectively, these data support and extend the role for the development of novel mGlu5 PAMs for the treatment of psychosis and cognitive deficits observed in individuals with schizophrenia.

Acute and chronic administration of immunomodulators induces anorexia in Zucker rats.

To investigate the possible involvement of leptin signaling in lipopolysaccharide (LPS) anorexia, we compared the anorectic effect of LPS in genetically obese (fa/fa) Zucker rats and in their lean (Fa/?) counterparts. The effects of interleukin-1beta (IL-1beta) and muramyl dipeptide (MDP) were also tested. LPS [100 microg/kg body weight (BW)], IL-1beta (2 microg/kg BW) and MDP (2.2 mg/kg BW) injected intraperitoneally (i.p.) at lights out reduced food intake similarly in obese and lean rats. LPS injection at 500 or 1000 microg/kg BW (i.p.) also reduced food intake and BW similarly in obese and lean rats, but obese regained BW faster than lean rats. LPS (2.45 microg or 9.8 microg/h/rat) administered chronically with i.p. implanted osmotic pumps reduced food intake similarly on experimental day 1, regardless of the genotype. After day 3, the lean rats' anorectic response and recovery were dose-dependent, whereas the anorectic response in obese rats was minimally affected by dose (significant dose effect only on day 3). Again, obese rats regained lost BW faster than lean rats. These results do not support a role for leptin as the sole mediator of anorexia induced by bacterial products (LPS and MDP) and IL-1beta.

A role for cyclooxygenase-2 in lipopolysaccharide-induced anorexia in rats.

Because nonselective cycloooxygenase (COX) inhibition attenuated anorexia after lipopolysaccharide (LPS) administration, we tested the ability of resveratrol (2.5, 10, and 40 mg/kg) and NS-398 (2.5, 10, and 40 mg/kg), selective inhibitors of the two COX isoforms COX-1 and -2, respectively, to attenuate LPS (100 microg/kg ip)-induced anorexia. NS-398 (10 and 40 mg/kg) administered with LPS at lights out attenuated LPS-induced anorexia, whereas resveratrol at all doses tested did not. Because prostaglandin (PG) E(2) is considered the major metabolite synthesized by COX, we measured plasma and cerebrospinal fluid (CSF) PGE(2) levels after LPS administration. LPS induced a time-dependent increase of PGE(2) in CSF but not in plasma. NS-398 (5, 10, and 40 mg/kg) blocked the LPS-induced increase in CSF PGE(2), whereas resveratrol (10 mg/kg) did not. These results support a role of COX-2 in mediating the anorectic response to peripheral LPS and point at PGE(2) as a potential neuromodulator involved in this response.

A role for serotonin in lipopolysaccharide-induced anorexia in rats.

Rats consistently reduce their food intake following injection of bacterial lipopolysaccharides (LPS). Because LPS increases CNS serotonin (5-HT) turnover, and because increases in CNS 5-HT turnover are associated with a decrease in food intake, we conducted a series of studies to examine 5-HT's potential role in LPS-induced anorexia. Chronic CNS 5-HT depletion by cisterna magna (CM) administration of 5,7-dihydroxytryptamine (5,7-DHT) failed to attenuate LPS-induced (100 microg/kg, ip) anorexia. In subsequent experiments, LPS was injected at lights out (hour 0) and [8-hydroxy-2-(di-n-propylamino)tetraline (8-OH-DPAT)] or N-CBZ-[(8beta)-1,6-dimethylergolin-8-yl]methylamine (metergoline) was injected at hour 5 - the time when LPS-treated rats become anorectic. Food intake was measured during the subsequent 2 h. In LPS-treated rats, 8-OH-DPAT (62.5, 125, or 250 microg/kg, sc) injection increased food intake. In a 2 x 2 factorial arrangement of LPS and 8-OH-DPAT, 125 microg/kg 8-OH-DPAT increased food intake significantly more in LPS-treated rats than in non-LPS-treated rats (significant LPS x 8-OH-DPAT interaction). In LPS-treated rats, 1 and 5 mg/kg metergoline significantly enhanced food intake. However, in a 2 x 2 arrangement of LPS and metergoline, 1 mg/kg metergoline failed to increase food intake in LPS and non-LPS-treated rats in two separate trials. The ability of the 5-HT(1A) receptor agonist 8-OH-DPAT to attenuate LPS-induced anorexia in rats supports a role of 5-HT in LPS-induced anorexia.

Inhibition of TNF-alpha production contributes to the attenuation of LPS-induced hypophagia by pentoxifylline.

Cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) are assumed to mediate anorexia during bacterial infections. To improve our understanding of the role that these two cytokines serve in mediating infection during anorexia, we investigated the ability of pentoxifylline (PTX), a potent inhibitor of TNF-alpha production, to block the anorectic effects of the bacterial products lipopolysaccharide (LPS) and muramyl dipeptide (MDP) in rats. Intraperitoneally injected PTX (100 mg/kg body wt) completely eliminated the anorectic effect of intraperitoneally injected LPS (100 microg/kg body wt) and attenuated the anorectic effect of a higher dose of intraperitoneally injected LPS (250 microg/kg body wt). Concurrently, PTX pretreatment suppressed low-dose LPS-induced TNF-alpha production by more than 95% and IL-1beta production 39%, as measured by ELISA. Similarly, high-dose LPS-induced TNF-alpha production was reduced by approximately 90%. PTX administration also attenuated the tolerance that is normally observed with a second injection of LPS. In addition, PTX pretreatment attenuated the hypophagic effect of intraperitoneally injected MDP (2 mg/kg body wt) but had no effect on the anorectic response to intraperitoneally injected recombinant human TNF-alpha (150 ug/kg body wt). The results suggest that suppression of TNF-alpha production is sufficient to attenuate LPS- and MDP-induced anorexia. This is consistent with the hypothesis that TNF-alpha plays a major role in the anorexia associated with bacterial infection.

Effect of sensory stimuli on huddling behavior of pigs.

Two experiments involving pigs at 1, 3, and 8 d of age were conducted to 1) compare huddling between littermates and nonlittermates, 2) study the ability of pigs to distinguish an anesthetized piglet from a piglet-shaped object, and 3) explore the importance of physical contact between pigs on huddling behavior. Experiments were conducted in an enclosed rectangular aluminum test chamber having pressure sensors beneath floor panels to detect test pig location. Test objects were placed on a platform at one end of the chamber and test pig location was monitored during a 45 min trial. Experiment 1 involved a total of 45 pigs (5 pigs/treatment on d 1, 2, and 3). The results indicate that, regardless of age (P > .05), when either a littermate or a nonlittermate occupied the platform, average location of test pigs that "settled" (ceasing to move for 7 min or more) was closer to the platform (P < .01), time spent near the platform was greater (P < .01), and movement about the chamber was less (P < .01) than when the platform was empty. No differences (P > .05) were observed between littermate and nonlittermate stimuli for these variables. During Exp. 2, the platform was covered with wire mesh. A total of 98 pigs were used in the study. Treatments were a cage containing 1) no object (n = 24), 2) a wooden block (n = 25), 3) a pig-shaped latex casting (n = 24), or 4) an anesthetized 8- to 10-d-old pig (n = 25). Pig age and treatment did not affect the percentage of time in each trial that pigs spent within 23.5 cm of the cage or the percentage of pigs settling within 23.5 cm of the cage. These studies show that pigs huddle similarly with littermates and nonlittermates and that physical contact with another piglet but not visual recognition of another piglet affects piglet huddling.

The effect of thermal environment and age on neonatal pig behavior.

Three experiments were conducted to evaluate the ability of a radiant environment and the presence of a littermate to attract pigs during the first 3 d of age. The effect of stimuli on pig movement was studied in an enclosed rectangular aluminum test chamber containing four similar sections that were heated independently. In Exp. 1, all sections were at 34.8 degrees C to evaluate the chamber for biases of where pigs located themselves at 1 (n = 24) and 2 d (n = 26) of age. More (P < .025) pigs settled (e.g., no movement for 7 min) in end sections than in middle sections. Age did not affect time to settle or settling location. The effect on pig location of heating one chamber end section to either 23, 40, 48, 56, or 64 degrees C and leaving the remaining sections unheated (24 degrees C) was determined in Exp. 2. Settling of pigs at 1 (n = 50) and 2 d (n = 50) of age was affected by temperature (P < .001) but not by age. The minimum distance between average pig location and the heated section occurred at 48 degrees C. Experiment 3 involved 15 pigs each at 1 and 3 d during a 1-h trial to compare the relative pig attraction to 1) a heated chamber end section at 44.4 degrees C when remaining sections were at 23.5 degrees C, 2) an anesthetized littermate in an end section when all sections were at 24.1 degrees C, or 3) a choice test involving a 45.5 degrees C end section and an anesthetized littermate in the opposite end section with three unheated sections at 23.7 degrees C. Average distance between the test animal and the heated section was greater (P < .01) than that between the test animal and an anesthetized pig. Pigs that were allowed a choice preferred to lie near an anesthetized littermate in a cold section rather than alone in a 45 degrees C section (P < .01), and they were less (P < .005) active when an anesthetized littermate was present in the chamber. Although radiant heat effectively attracted pigs, heat was less attractive than an anesthetized littermate. The greater attraction of pigs to a littermate than to radiant heat may explain why pigs remain near the sow and littermates during d 1 and 2 after birth.

Lysine deficiency alters diet selection without depressing food intake in rats.

Under states of protein deficiency, the dietary limiting amino acid, rather than protein content, can act as the dietary stimulus to control diet selection. If fact, threonine-deficient rats will alter their diet selection patterns solely on the basis of very small changes (0.009 g/100 g) in the dietary threonine concentration. In these studies, we assessed whether lysine-deficient rats will also alter their diet selection patterns on the basis of small changes in dietary Lys concentration. In all experiments, growing rats were adapted to diets in which the protein fraction (purified amino acids or wheat gluten) was limiting in Lys. They were then given a choice between the adaptation diet (AD) diet and a slightly more deficient diet. Rats that were adapted to a Lys-deficient diet (0.25 g Lys/100 g) selected their AD over diets containing as little as 0.01% less Lys (P < 0.01) within 5 d. To determine how deficient rats must be before they alter their selection patterns, rats were adapted to diets containing various levels of Lys, i.e., 2 levels below the requirement for growth and 2 levels above the requirement for growth, but below the requirement for maximal nitrogen retention. Only rats adapted to diets containing Lys below their requirement for growth selected their AD over a diet containing 0.05% less Lys (P < 0.005). Finally, to determine whether rats will alter their selection to whole protein-based diets, rats were adapted to 25% wheat gluten diets supplemented with 0.03-0.21% Lys. Rats selected the AD over a diet containing as little as 0.09% less supplemental Lys by d 4 of the trial (P < 0.05). We conclude that rats are sensitive to changes as small as 0.01% in dietary Lys concentration, but that sensitivity requires prior adaptation to Lys-deficient diets.

Vagal and splanchnic afferents are not necessary for the anorexia produced by peripheral IL-1beta, LPS, and MDP.

We investigated the extrinsic gut neural mediation of the suppression of food intake in male Sprague-Dawley rats induced by peripheral intraperitoneal administration of 2 microg/kg interleukin-1beta (IL-1beta), 100 microg/kg bacterial lipopolysaccharide (LPS), and 2 mg/kg muramyl dipeptide (MDP). Food intake during the first 3 and 6 h of the dark cycle was measured in rats with subdiaphragmatic vagal deafferentation (n = 9), celiac superior mesenteric ganglionectomy (n = 9), combined vagotomy and ganglionectomy (n = 9), and sham deafferentation (n = 9). IL-1beta, LPS, and MDP suppressed food intake at 3 and 6 h in all surgical groups. The results demonstrate that neither vagal nor nonvagal afferent nerves from the upper gut are necessary for the feeding-suppressive effects of intraperitoneal IL-1beta, LPS, or MDP in the rat and suggest that peripheral administration of immunomodulators produces anorexia via a humoral pathway.

The effect of farrowing crate heat lamp location on sow and pig patterns of lying and pig survival.

Three experiments were conducted to study sow and pig behavior during the 1st 3 d after birth and pig survival during the 1st 2 wk after farrowing. In Exp. 1, 23 sows were housed in conventional farrowing crates that were divided into five sections: a .5- x 1.5-m front creep section and the remaining area divided into four sections, .75 x 1.05 m each. Air temperature was maintained at 19 degrees C, and a 250-W heat lamp was placed at the right side of the front creep in Treatment 1 (T1), or in the creep at the right side of the sow for Treatment 2 (T2). The percentage of pigs within 8 cm of the sow's trunk was not affected by treatment, but it decreased (P < .001) from 61.8 +/- 3.4% on d 1 to 28.1 +/- 3.5% on d 3. As the percentage of pigs near the sow decreased, the percentage of pigs within the section containing the heat lamp increased (T1, P < .05; T2, P < .10). Experiment 2 involved 15 sows and litters housed as in Exp. 1, except that heat lamps were not provided, and average air temperature was 27.3 +/- .2 degrees C during behavioral observations. Even though the portion of the litter near the sow decreased (P < .001) from d 1 to d 3 (d 1, 57.0 +/- 3.4%; d 2, 42.9 +/- 3.3%; d 3, 31.7 +/- 3.3%), pigs did not concentrate in any specific section as they moved away from the sow. The average number of pigs within the front creep section (Section 1) for the 3-d period was less than (P < .01) the number in any other crate section. Experiment 3 involved 147 sows and tested the effect of solid creep floor covering on pig survival for each of the heat lamp locations used in Exp. 1. Neither heat lamp location nor floor covering affected pig survival. During the 1st 3 d of life, pigs tend to lie near the sow regardless of heat lamp location or air temperature. Heat lamp position and floor covering under the lamp do not affect pig survival.

Small changes in essential amino acid concentrations alter diet selection in amino acid-deficient rats.

The rat's sensitivity to changes in the dietary limiting amino acid concentration (LAA) was examined on the basis of dietary selection. Rats were adapted to purified low protein basal (Basal) diets in which threonine (Thr) was the LAA (0.188-0.212% wt/wt of diet). In Experiment 1, rats made a clear selection for their adaptation diet over a diet containing 0.012% less threonine after 2-3 d of choice. Rats made no clear dietary selection when given a choice between their adaptation diet and a diet containing 0.012% more threonine. Experiment 2 was conducted to examine the rat's sensitivity to small decreases in the LAA concentration. Rats adapted to a 0.200% Thr-Basal diet clearly responded to decreases as small as 0.009% in the concentration of threonine and selected against the more deficient diet when given a choice between it and the 0.200% Thr-Basal adaptation diet. Because plasma and brain amino acid concentrations are important for detection of other amino acid deficiencies, these variables were measured to determine whether they were affected by such small changes in dietary amino acid concentration. In Experiments 3 and 4, rats were adapted to the 0.200% Thr-Basal diet and then fed 0.188, 0.200 or 0.212% Thr-Basal diets for 6 h, or 0.188 and 0.212% Thr-Basal for 54 h. Amino acid concentrations in plasma, prepiriform cortex and anterior cingulate cortex were not significantly different among treatments. Norepinephrine concentration in the prepiriform cortex was not affected by dietary treatment. We conclude that small decreases in LAA concentration can cause selection against the more deficient diet, but that detection of such deficiencies does not require significant changes in plasma and brain amino acid concentrations.

Ovariectomy and estradiol affect postingestive controls of sucrose licking.

Ovariectomy (OVX) has been shown to increase, and estradiol replacement to decrease, meal size in rats. Because little is known about how estradiol influences meals, we conducted two experiments to examine the effects of OVX and beta-estradiol 3-benzoate (EB) replacement on the microstructure of licking behavior. In both experiments, patterns of licking were analyzed in adult female Sprague-Dawley rats during an 0.8 M sucrose test meal. In Experiment 1, meal microstructure was determined preOVX and 10-12 days postOVX. Rate of licking following OVX was not changed during min 1 of the meal, but was significantly faster during min 2-4 of the meal (p < 0.03). The numbers of bursts (runs of licks separated by 250-500 ms) and numbers of clusters (runs of licks separated by > 500 ms) were significantly increased during min 2-4 (p < 0.05). In Experiment 2, OVX rats received EB replacement. Rate of licking after EB replacement was not changed during min 1 of the meal, but was significantly slower during the remainder of the meal (min 2-4, min 5-7, and min 8-10). Burst size, cluster size, and interburst interval were less after EB replacement during min 5-7 of the test meal (all p < 0.05). Because both OVX and EB replacement failed to alter the rate of licking during min 1, estrogen did not appear to alter the palatability of sucrose. OVX and EB replacement did appear to affect a postingestive mechanism(s) that is engaged within 2-4 min of meal onset.

Rats on a macronutrient self-selection diet eat most meals from a single food cup.

Computerized meal pattern analysis was performed on female Sprague-Dawley rats (n = 10). Rats were housed for 14 days in cages adapted for macronutrient selection. Rats selected their diet from vitamin and mineral supplemented, semipurified sources of carbohydrate, fat and protein, in three individual food cups. For analysis of meal patterns, minimum meal size was > 50 mg, and mean minimum intermeal interval was 7.9 min. Daily energy intake averaged 289.0 kJ, with 75% occurring during the dark cycle. Energy intake was 28% carbohydrate, 50% fat and 22% protein. Of 12.3 daily meals, 56% were from one, 35% from two, and only 9% of the meals were from three food cups. Seventy percent of the time carbohydrate was the first meal of the dark cycle. The average number of meals per day eaten from carbohydrate, fat, and protein were not significantly different (6.9, 6.2 and 5.6, respectively, p > 0.05). Energy intake for a meal was greatest when fat was eaten (18.35 kJ), than when either carbohydrate (8.68 kJ) or protein (8.97 kJ) was eaten. Meal duration was 7.03 min for carbohydrate, 3.75 min for fat, and 7.47 min for protein. These results provide evidence that rats on a macronutrient self-selection diet eat most meals from a single macronutrient source.

Timing and dose of amino acids injected into prepyriform cortex influence food intake.

The effects of time before feeding and dose of dietary-limiting amino acids (DLAA) injected into the prepyriform cortex (PPC) on intake of amino acid-imbalanced diets were evaluated. Intake of imbalanced diet was increased from approximately 50% to approximately 75% of baseline when an optimal amount of DLAA (1 nmol L-isoleucine or 2 nmol L-threonine) was injected immediately prior to feeding. Injections made several hours prior to feeding were more effective, increasing intake of imbalanced diets to approximately 85% of baseline. Delivering two half-optimal doses of DLAA, several hours apart, increased intake of imbalanced diet only to the same level as a single injection of the optimal dose immediately prior to feeding. The increase in intake of a threonine-imbalanced diet after injecting 2 nmol threonine 6 h prior to feeding was abolished if an additional 2 nmol threonine was injected immediately prior to feeding. It appears that it is the sum of the changes in tissue DLAA concentrations in the PPC that are recognized and influence food intake when amino acid imbalanced diets are fed.

ICS 205-930 and feeding responses to amino acid imbalance: a peripheral effect?

Serotonin3 (5-HT3) receptor antagonists (ICS 205-930 and MDL 72222) have been shown to block or ameliorate the anorectic response of the rat to amino acid imbalanced (IMB) diets. Two experiments were conducted to determine whether the effects of these antagonists are mediated through central or peripheral 5-HT3 receptors. In Experiment One, ICS 205-930 (ICS) was injected centrally, into either the lateral ventricle (doses: 0.3 pmol to 10 nmol), or the cisterna magna (62 nmol). The intake of rats fed an isoleucine IMB diet was not affected by these injections. In Experiment Two, rats received an IP injection of either saline, ICS, or a quaternized derivative of ICS (Q-ICS) that should not cross the blood-brain barrier. Both ICS- and Q-ICS-injected rats ate significantly more (p less than 0.05) IMB diet than saline-injected rats. Intake of IMB diet was not different (p greater than 0.4) between ICS and Q-ICS groups. From these results, it appears that ICS restores intake of IMB through a peripheral component.

Distribution of dietary limiting amino acid injected into the prepyriform cortex.

Diffusion or metabolism of the dietary limiting amino acid (DLAA) in the prepyriform cortex (PPC) may account for the time lag between injection of the DLAA into the PPC and the increase in intake of an amino acid-imbalanced diet. Results from the injection of [3H]Leu +/- [14C]Thr (DLAA) into the PPC indicated rapid (less than 15 min) and limited diffusion (85-90% of recovered label was less than or equal to 1 mm from the injection site). 3H and 14C decreased in the trichloroacetic acid (TCA)-soluble fraction and increased in the TCA-insoluble fraction during the first 1.5 h and remained constant in the TCA-insoluble fraction 1.5-6 h after injection. An increase (approximately 50%) in 3H in the TCA-insoluble fraction was found less than or equal to 30 min after injection of the DLAA. There was no affect of the DLAA on 3H in the TCA-soluble fraction. These results indicated that a change in metabolism within the PPC may be responsible for the delay in onset of the feeding response after injection of the DLAA into the PPC.