Mu-opioid receptor cellular function in the nucleus accumbens is essential for hedonically driven eating.

Acute pharmacological studies have implicated mu-opioid receptors (MORs) in the shell of the nucleus accumbens (NAC) in mediating responses for palatable food and other natural and drug-induced rewards. However, the long-term behavioral effects of inactivating signal transduction via accumbal MORs, as quantified by an anatomically defined loss of cellular activity, have never been analysed. We combined microinfusion of the irreversible MOR antagonist, beta-funaltrexamine (beta-FNA; 8.0 nmol/0.8 microL, n=9; controls, n=6) with mapping by [35S]GTPgammaS autoradiography to demonstrate an anatomically specific loss of the coupling of MORs to their G-proteins in the dorsal caudomedial shell of the NAC in rabbits. beta-FNA did not alter the stimulated coupling of kappa-opioid receptors. This selective blockade of the cellular function of MORs persistently decreased consumption of a palatable sucrose solution by 40% during a daily 4-h test conducted 2, 3 and 4 days after infusion. beta-FNA did not alter body weight or 20-h consumption of standard chow or water. In 10 different rabbits, infusion of the selective, competitive MOR antagonist, CTAP (D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2) into the same locus produced a reversible decrease in sucrose consumption, with normal intakes returning on the next day. Together, these data appear to establish that MORs in this accumbal subregion support responding for orosensory reward. Overall, these results visualize a discrete brain locus where cellular actions of endogenous opioids mediate behaviors involved in self-administration of foods and perhaps other hedonically valued substances, such as ethanol and drugs of abuse.

A 5-HT2C agonist elicits hyperactivity and oral dyskinesia with hypophagia in rabbits.

Serotonergic 5-HT2C and 5-HT1B receptors mediate inhibitory controls of eating. Questions have arisen about potential behavioral and neurological toxicity of drugs that stimulate the 2C site. We evaluated eating and other motor responses in male Dutch-belted rabbits after administration of m-chlorophenylpiperazine (mCPP). Studies conducted in vitro and in vivo assessed the pharmacological specificity of the ingestive actions of this agent. mCPP (0.15-10 micromol/kg sc) reduced consumption of chow and 20% sucrose solution with equal potencies (ED50 approximately equal 0.6 micromol/kg). In radioligand binding to rabbit cortex, mCPP displayed 15-fold higher affinity for 5-HT2C than for 5-HT1B receptors. The serotonin antagonist mesulergine (7000-fold selective for 5-HT2C) reversed the hypophagic action of mCPP, but the 5-HT1B/1D antagonist GR127,935 did not. GR127,935 (0.5 micromol/kg) did prevent hypophagia produced by the highly selective 5-HT1B/1D agonist GR46,611. Observational methods demonstrated that mCPP decreased the frequency of eating chow but increased other motor activities. When rabbits consumed sucrose, videoanalysis revealed that mCPP reduced total time licking and the duration of individual bouts, but not bout frequency or the actual rate of consumption. mCPP increased locomotor and other activities, and greatly increased vacuous oromotor stereotypies and tongue protrusions. Nonetheless, rabbits licked accurately at the spout for sucrose. When sucrose was infused intraorally through a cheek catheter, mCPP actually increased the peak amplitude and overall magnitude of jaw movements. We conclude that mCPP stimulates 5-HT2C receptors to reduce food intake in rabbits. This hypophagia involves disruption of appetitive components of eating and is accompanied by adverse motor actions. This profile raises questions about the use of the 5-HT2C receptor as a target for novel therapeutic agents for obesity.

Neuropeptide FF exerts pro- and anti-opioid actions in the parabrachial nucleus to modulate food intake.

Neurons that synthesize the morphine modulatory peptide neuropeptide FF (NPFF; Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2) densely innervate the parabrachial nucleus (PBN), an area implicated in regulating food intake. We analyzed opioid-related actions of NPFF in feeding in adult male Sprague-Dawley rats. Unilateral infusion of 2 nmol/0.5 microl of the mu-opioid receptor agonist [d-Ala2,NMe-Phe4,glycinol5]enkephalin (DAMGO) into the lateral PBN increased 4-h food intake from 0.7 +/- 0.1 to 3.3 +/- 0.3 g. NPFF (1.25-5.0 nmol) prevented this hyperphagic mu-opioidergic action. In rats fed after 4-h deprivation (baseline = 12.3 +/- 0.3 g/2 h), 5 nmol of NPFF did not alter and larger doses (10 and 20 nmol) actually increased food intake (+36, 54%). Twenty nanomoles also elevated intake of freely feeding rats (from 0.7 +/- 0.1 to 5.1 +/- 1.0 g/4 h). The opioid receptor blocker naloxone (10 nmol) antagonized this increase. These data reveal both pro- and anti-opioid actions of NPFF in the PBN to modulate feeding. The mechanisms for the opposite actions of low and high concentrations of this neuropeptide in parabrachial regulation of food intake remain to be determined.

An orexigenic role for mu-opioid receptors in the lateral parabrachial nucleus.

The pontine parabrachial nucleus (PBN) has been implicated in regulating ingestion and contains opioids that promote feeding elsewhere in the brain. We tested the actions of the selective mu-opioid receptor (mu-OR) agonist [d-Ala2,N-Me-Phe4,Gly5-ol]enkephalin (DAMGO) in the PBN on feeding in male rats with free access to food. Infusing DAMGO (0.5-4.0 nmol/0.5 microl) into the lateral parabrachial region (LPBN) increased food intake. The hyperphagic effect was anatomically specific to infusions within the LPBN, dose and time related, and selective for ingestion of chow compared with (nonnutritive) kaolin. The nonselective opioid antagonist naloxone (0.1-10.0 nmol intra-PBN) antagonized DAMGO-induced feeding, with complete blockade by 1.0 nmol and no effect on baseline. The highly selective mu-opioid antagonist d-Phe-Cys-Trp-Arg-Thr-Pen-Thr-NH2 (CTAP; 1.0 nmol) also prevented this action of DAMGO, but the kappa-antagonist nor-binaltorphimine did not. Naloxone and CTAP (10.0 nmol) decreased intake during scheduled feeding. Thus stimulating mu-ORs in the LPBN increases feeding, whereas antagonizing these sites inhibits feeding. Together, our results implicate mu-ORs in the LPBN in the normal regulation of food intake.

Parabrachial infusion of D-fenfluramine reduces food intake. Blockade by the 5-HT(1B) antagonist SB-216641.

Systemic administration of the serotonin (5-HT) releaser/reuptake inhibitor, D-fenfluramine decreases consumption of food in mammals. This hypophagic action involves loci at several levels of the neuraxis. Indirect evidence implicates the parabrachial nucleus (PBN) of the pons as one of these regions. Consistent with this hypothesis, unilateral infusion of D-fenfluramine (200, 280, and 400 nmol/0.5 microl) directly into the lateral PBN (LPBN) of male rats reduced food intake by 33%, 56%, and 66% from baseline (7.3 +/- 0.7 g) during a 30-min test with chow. Infusions lateral, medial, and dorsal to the PBN were ineffective. Stimulating 5-HT(1B) receptors in the PBN also reduces feeding. Administration of the selective 5-HT(1B) agonist CP-93,129 (3-(1,2,5,6-tetrahydropyrid-4-yl)pyrrolo[3,2-b]pyrid-5-one) (0, 0.625, 2.5, and 10 nmol/0.5 microl) into the PBN reduced food intake by 25--79%. The selective 5-HT(1B) antagonist SB-216641 (N-[3-[3-(dimethylamino(ethoxy]-4-methoxyphenyl]-2'-methyl-4'-(5-methyl-1,2,4-oxadiazol-3-yl)-[1,1'-biphenyl]-4-carboxamide) (2.5 nmol) completely blocked the hypophagic action of the approximate ED(50) doses of CP-93,129 (2.5 nmol) and D-fenfluramine (280 nmol). These data strongly suggest that directly or indirectly activating 5-HT(1B) receptors in the LPBN inhibits feeding and implicates this pontine region in the serotonergic regulation of eating and satiation.

Anatomical but not functional recovery from a sciatic nerve crush is enhanced by treatment with testosterone.

PURPOSE: Until recently, there has been a limited amount of research comparing functional and anatomical recovery following nerve injury. Previous studies emphasizing anatomical recovery (such as axonal number) have shown that testosterone promotes regeneration in crushed and transected nerves. The purpose of this study was to assess the effect of testosterone on the functional recovery of the sciatic nerve follow-ing a unilateral crush injury. METHODS: Young adult male Sprague-Dawley rats were injected daily with either 500 micro g testosterone proprionate or vehicle alone. The recovery course was followed for six weeks using functional and behavioral testing. Behavioral tests included a footprint gait analysis (as a measure of motor function), response to a skin pinch, and warm water withdrawal (measures of sensory function). RESULTS: Immediately following surgery, all tests indicated complete denervation to the leg distal to the crush site. Anatomical analysis revealed a 22 % increase in the number of axons in testosterone treated animals at 6 weeks post-crush, but no indication of differences in functional recovery. The results of behavioral testing indicated only minor differences in functional recovery as a result of testosterone treatment. CONCLUSION: The results indicate the need for a detailed comparison between anatomical regeneration and functional recovery. An increase in axon number alone may not be an accurate indicator of successful regeneration.