Search

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.

Ingestion of imbalanced amino acid diets (IMB) has been associated with a decrease in norepinephrine (NE) concentration in the prepyriform cortex (PPC), an area essential for the anorectic response to IMB. Decreased NE could result from activity-induced release (and subsequent metabolism) of the transmitter. If activity of the NE system is important in the rat's anorectic response to IMB, reduced NE activity should result in increased IMB intake. Therefore, the feeding response to IMB was measured after injecting clonidine (Clon) into the PPC to inhibit NE release. At 3 and 6 h after Clon (1.0 and 1.5 micrograms/rat) injections, IMB intake was increased from 69 (the usual response to IMB in untreated animals) to > 100% of control intake. Effective injection sites did not include the gustatory neocortex, an area important for conditioned taste aversions. Thus activation of the NE system in the PPC may be associated with the initial reduced intake of IMB, suggesting that NE activity in the PPC has a role in the neural mechanisms that subserve recognition of amino acid deficiency.

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.

The effect of inhibiting protein synthesis within the prepyriform cortex (PPC) on intake of an amino acid imbalanced diet was evaluated in rats receiving bilateral injections of the dietary limiting amino acid (DLAA). Injection of the DLAA into the PPC increased intake of the imbalanced diet by 150% and incorporation of [3H]leucine into the trichloroacetic acid insoluble fraction of PPC homogenates by 248%. Coinjection of puromycin (100 mumol/L) or actinomycin D (10 mumol/L) blocked the increase in intake of the imbalanced diet but had no effect on intake of the basal diet. Puromycin blocked the increase in intake of the imbalanced diet whether injected with the DLAA or 6 h later. Selection of the imbalanced diet over a protein-free diet when the DLAA was administered was blocked by co-injecting puromycin or actinomycin D. The results indicated that the increased intake and the reversal of aversion to the imbalanced diet when the DLAA was injected into the PPC required de novo protein synthesis. The proteins necessary for the feeding response seemed to have a short half-life and seemed to require synthesis of new mRNA. We conclude that changes in concentration of the DLAA within the PPC influenced protein synthesis at the genomic level.

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.

The role of adrenal function in the anorectic response and adaptation of rats to a diet with an isoleucine (Ile) imbalance was investigated. In the first of four experiments, rats were fed a mildly Ile-imbalanced diet after treatment with metyrapone, and inhibitor of glucocorticoid synthesis. In two separate experiments, rats were presented with either a mildly or severely Ile-imbalanced diet (4.93 and 9.86% imbalanced amino acid mixture, respectively) after bilateral adrenalectomy. Finally, the effects of ICS 205-930, a serotonin-3 receptor antagonist, on the intake of mildly Ile-imbalanced diet were tested in adrenalectomized animals. In each experiment a 2 X 2 factorial design was used. Neither metyrapone nor adrenalectomy altered the initial depression in the intake of an imbalanced diet. The adaptation phase in the response of adrenalectomized rats fed a mildly Ile-imbalanced diet was not different from that of controls, but adrenalectomized rats fed severely Ile-imbalanced diets were unable to adapt. Adrenalectomy did not alter the anti-anoretic activity of ICS 205-930 in this model. These results suggest that adrenal hormones are not necessary for the initial anoretic response or adaptation of rats to an Ile-imbalanced diet, nor are they implicated in the anti-anorectic effect of serotonin-3 blockade.

Microinjection of the dietary limiting essential amino acid (DLAA) into the prepyriform cortex (PPC) increased intake of a diet having an imbalance among the essential amino acids (imbalanced diet) from 50-55% of baseline, when artificial cerebrospinal fluid (aCSF) was injected, to 70-75% of baseline. The increase in intake of the imbalanced diet by DLAA injection became apparent after 3-6 h and was maintained throughout the dark period. Meal size, meal duration, and the number of meals returned to normal after bilateral injections of the DLAA into the PPC of rats fed the imbalanced diet. Injection of the DLAA 30 min before the onset of the dark phase increased intake of imbalanced diet to 70% of baseline intake. When injections of threonine or isoleucine were made 6 and 3 h, respectively, prior to onset of the dark phase, intake of imbalanced diet increased to 85% of baseline intake. Results suggest that some form of processing of the injected DLAA within the PPC is necessary to increase the intake of imbalanced diets.

The mechanisms underlying the reduced intake of an amino acid-imbalanced diet (imbalanced diet) appears to involve a decrease in the content of the dietary limiting amino acid (DLAA) in the prepyriform cortex (PPC). Intake of imbalanced diet was increased from 45-50 to 70-75% of baseline after bilateral injection of the DLAA directly into the PPC, following an inverted U-shaped dose-response curve. Injections had no effect on intake of basal diets. Injection of the DLAA into the PPC reversed the aversion to imbalanced diet in choice studies, as rats selected an imbalanced diet over protein-free diet after such injections. Intake of imbalanced diet did not increase after a nonlimiting amino acid was injected into the PPC or after injections of the DLAA into other brain areas. Results were similar when either threonine or isoleucine was the DLAA. These results confirm that the decrease in the concentration of the DLAA in the PPC is involved in the reduction in intake of imbalanced diets.

The role of serotonin3 (5-HT3) receptors in the initial food intake depression of rats ingesting amino acid imbalanced or high-protein diets was investigated. The 5-HT antagonists metergoline, pirenpirone, ICS 205-930, and MDL 72222, the dopamine antagonist pimozide, or the alpha-adrenergic antagonist phentolamine were injected 15-45 min before presentation of test diets. Food intake was measured at intervals for 3 days. The 5-HT3 antagonists, ICS 205-930 and MDL 72222, restored feeding of a mild isoleucine (Ile)-imbalanced diet to control levels, although MDL 72222 had a longer time course of action. ICS 205-930 also increased intake of a severe Ile-imbalanced diet and Thr-imbalanced diet but not a high-protein (44% casein) diet. Treatment with metergoline, which blocks 5-HT1, 5-HT2, and dopamine receptor sites but not 5-HT3 sites, increased intake of the basal diet at 3 and 6 h but did not significantly alter intake of the mild Ile-imbalanced diet. Although pimozide tended to increase intake of the mild imbalanced diet, neither dopamine nor alpha-adrenergic receptor antagonism significantly affected imbalanced diet intake. Thus 5-HT3 receptors may mediate the anorexigenic activity of 5-HT associated with feeding an amino acid-imbalanced diet.