Malcolm K. McGowan

Chronic intrahypothalamic infusions of insulin or insulin antibodies alter body weight and food intake in the rat

In Experiment 1, one-week infusion of insulin (0.15, 1.5, or 15.0 microU/hr) into the ventromedial hypothalamus (VMH) of rats reduced body weight (BW) and nighttime food intake (FI). While 0.15 microU/h decreased daytime FI, 1.5 microU/h increased daytime FI and 15.0 microU/h left daytime FI unchanged. Total daily FI was decreased by the two highest doses. In Experiment 2, intra-VMH infusion of specific insulin antibodies (1.5 microUeq/h) increased BW and FI, while C-peptide antibodies were ineffective. In Experiment 3a, intracerebroventricular infusions of insulin failed to decrease FI and BW comparably to similar intrahypothalamic infusions. In Experiment 3b, intra-VMH insulin was infused via cannulae that bypassed the cerebral ventricles. The decrease in FI and BW was comparable to that observed when insulin was infused via cannulae that penetrated a ventricle. Histology from animals used in Experiments 1-3 indicates that optimum sites for insulin-induced changes in BW and FI in the hypothalamus lie in an area that includes portions of the paraventricular, arcuate, dorsomedial, and ventromedial nuclei.

Effects of chronic intrahypothalamic infusion of insulin on food intake and diurnal meal patterning in the rat.

In Experiment 1, rats were chronically infused with insulin (2.7, 27, or 270 ng/hr) or 0.9% saline into the ventromedial (VMH), medial perifornical (PF), or lateral (LH) hypothalamus. VMH infusions of insulin caused a significant, dose-dependent decrease in food intake and body weight; PF infusion of insulin was less effective, but significant; whereas LH infusions of insulin were ineffective. In Experiment 2, rats were chronically infused with insulin (0.54 ng/hr) or 0.9% saline into the VMH, paraventricular (PVN), or posterior (PN) hypothalamic nucleus. Subjects that received VMH or PN infusions of insulin failed to regain weight lost as a result of surgery even 2 weeks after infusion; subjects that received PVN infusions of insulin regained their preoperative weights faster than did controls. All of the groups that received insulin significantly increased their daytime food intake during the infusion period and decreased their night food intake slightly; 24-hr food intake remained unchanged.

Antinociception produced by microinjection ofl-glutamate into the ventromedial medulla of the rat: mediation by spinal GABAA receptors

This study examined whether the antinociception produced by glutamatergic stimulation of neurons in the nucleus raphe magnus (NRM) or nucleus reticularis gigantocellularis pars alpha (NGCp alpha) is mediated by activation of GABAA receptors in the spinal cord. Two approaches were used. The first approach determined the ability of intrathecally (i.t.) administered bicuculline, a competitive GABAA receptor antagonist, to attenuate the antinociception produced by microinjection of L-glutamate in the NRM or NGCp alpha. Bicuculline was selected on the basis of an initial study that determined that 0.3 micrograms i.t. bicuculline caused a 3.4-fold rightward shift in the dose-effect relationship of the i.t.-administered GABAA agonist, isoguvacine, without producing allodynia or alterations in nociceptive threshold. The GABAA antagonist SR 95531 was judged unsuitable because it caused only a modest 1.7-fold rightward shift in the dose-effect relationship of isoguvacine at doses that did not produce allodynia. The second approach determined the ability of i.t. administered diazepam, a benzodiazepine receptor agonist, to enhance the antinociception produced by microinjection of L-glutamate in these nuclei. Diazepam was selected because of its ability to enhance the actions of GABA at the GABAA receptor. Rats were pretreated with i.t. administration of 0.3 micrograms bicuculline, 40 micrograms diazepam, or vehicle, after which 30 nmol L-glutamate was microinjected into the NRM or NGCp alpha. Microinjection of L-glutamate into the NRM or NGCp alpha in vehicle-pretreated rats significantly increased tail flick latency. The antinociception produced by microinjection of L-glutamate in the NGCp alpha was antagonized by bicuculline and enhanced by diazepam. In contrast, the antinociception evoked from sites in the NRM was only partially attenuated by bicuculline and was not enhanced by diazepam. In an ancillary experiment, i.t. administration of 0.3 micrograms bicuculline or 40 micrograms diazepam did not alter tail skin temperature or nociceptive threshold, suggesting that their effects on glutamate-induced antinociception were not secondary to alterations in tail skin temperature. Taken together, these results support the hypothesis that the antinociception produced by activation of neurons in the NGCp alpha, but not the NRM, is mediated in part by an action of GABA at GABAA receptors in the spinal cord.

Intrathecal GABAB antagonists attenuate the antinociception produced by microinjection ofl-glutamate into the ventromedial medulla of the rat

This study examined whether the antinociception produced by glutaminergic stimulation of neurons in the nucleus raphe magnus (NRM) and nucleus reticularis gigantocellularis pars alpha (NGCp alpha) is mediated by an action of GABA at GABAB receptors in the spinal cord. Rats were pretreated with intrathecal (i.t.) administration of the selective GABAB receptor antagonists phaclofen (100 micrograms) or CGP 35348 (30 micrograms), the serotonin receptor antagonist methysergide (30 micrograms), or vehicle. Fifteen min later, 30 nmol L-glutamate was microinjected into the NRM, NGCp alpha, or sites in the medulla outside these two regions. Microinjection of L-glutamate into the NRM or NGCp alpha in vehicle-pretreated rats significantly increased tail flick latency. This increase was antagonized, but not abolished, by i.t. pretreatment with 30 micrograms CGP 35348 or 100 micrograms phaclofen. Pretreatment with 30 micrograms methysergide completely antagonized the antinociception produced by L-glutamate. Microinjection of L-glutamate at medullary sites outside the NMR or NGCp alpha did not produce antinociception. In an ancillary experiment, the possibility that the ability of methysergide, phaclofen or CGP 35348 to antagonize glutamate-induced antinociception was related to non-specific increases in tail skin temperature was explored. Although phaclofen or methysergide increased tail skin temperature, the magnitude and time course of this increase were not consistent with the antagonism of glutamate-induced antinociception. Moreover, administration of CGP 35348 resulted in a modest decrease in tail skin temperature. Thus, antagonism of glutamate-induced antinociception does not appear to result from non-specific alterations in tail skin temperature.(ABSTRACT TRUNCATED AT 250 WORDS)

Antinociception produced by systemic R(+)-baclofen hydrochloride is attenuated by CGP 35348 administered to the spinal cord or ventromedial medulla of rats

This study examined the sites in the central nervous system at which subcutaneously-administered R(+)-baclofen hydrochloride (baclofen), the most active isomer of this prototypic gamma-aminobutyric acid (GABA)B receptor agonist, acts to produce antinociception in the rat. To determine whether baclofen acts in the spinal cord, either saline or the GABAB receptor antagonist CGP 35348 was injected intrathecally in rats pretreated 24 min earlier with 1 or 3 mg/kg s.c. baclofen. Intrathecal (i.t.) injection of 3 or 10 micrograms of CGP 35348 antagonized the increase in tail-flick and hot-plate latency produced by either dose of baclofen. To determine whether baclofen acts at sites in the ventromedial medulla (VMM), either saline or CGP 35348 was microinjected in the nucleus raphe magnus or nucleus reticularis gigantocellularis pars alpha of rats pretreated 24 min earlier with 1 or 3 mg/kg s.c. baclofen. Microinjection of 0.5 or 3 micrograms of CGP 35348 at sites in the VMM produced at best only a very modest attenuation of the antinociceptive effects of baclofen. These data suggest that systemically-administered baclofen acts at sites in both the spinal cord and the VMM, but that its antinociceptive effects are likely to be mediated to a greater extent by a spinal, rather than medullary site of action. However, a definitive comparison of the relative contribution of GABAB receptors in these two regions is precluded by differences in the diffusion and concentrations of the antagonist in the spinal cord and brainstem. Finally, microinjection of 0.5 or 3.0 micrograms of CGP 35348 in the nucleus raphe magnus or nucleus reticularis gigantocellularis pars alpha of saline-pretreated rats did not alter tail-flick or hot-plate latency. This finding suggests that, unlike GABAA receptors, GABAB receptors do not mediate the tonic GABAergic input to neurons in these nuclei.

Depletion of neurons from preoptic area impairs drinking to various dipsogens

We depleted neurons from the lateral preoptic region (LPO) of rats by iontophoretic application of the neurotoxin kainic acid (KA). Rats with KA-induced damage to LPO neurons drank less water than controls after subcutaneous (SC) administration of: (a) 5 ml of a 1.0 M saline solution or (b) 5 ml of a 30% polyethylene glycol (PG) solution. The drinking response to 1.5 mg/kg angiotensin II (AII) was significantly smaller than was that of controls 15 min but not 1 hour after the SC injection. The experimental animals drank as much water as controls under ad lib conditions and showed normal drinking responses to low concentrations of hypertonic saline (0.5 M) or PG (10% or 20%).

Lesions of the MPO or AV3V: influences on fluid intake

Electrolytic lesions in the MPO of rats had no significant effects on ad lib food and water intake, but impaired the drinking response to 1 M NaCl. Large MPO lesions also produced a persistent increase in plasma osmolality. In Experiment 2, we depleted neurons from the MPO of rats by iontophoretic application of the neurotoxin kainic acid (KA) which destroys nerve cell bodies without damage to fibers of passage. KA-induced neuron depletion in the MPO of rats significantly reduced the drinking response to 1.0 M saline, to 30% PG, and to 30 micrograms/kg isoproterenol. Ad lib water intake and drinking responses to food or water deprivation, to low concentrations (0.5 M) of hypertonic saline, to low concentrations (10% or 20%) of PG, and to systemic administration of 1.5 mg/kg angiotensin II were within the normal range. In Experiment 3, rats with electrolytic lesions that were strictly confined to the tissue immediately surrounding the wall of the anteroventral portion of the third ventricle (AV3V), without invading the MPO displayed normal ad lib food and water intake and plasma osmolality as well as drinking responses to water deprivation, hypertonic saline (0.5 or 1.0 M), angiotensin II (1.5 mg/kg) and isoproterenol (30 micrograms/kg).

Water intake during chronic preoptic infusions of osmotically active or inert solutions

To further elucidate the role of the lateral preoptic area (LPO) as an osmoreceptive region, rats received chronic infusions (2 weeks) of low volumes (0.5 microliters/h) solutions of hypertonic sodium chloride (NaCl; 0.16 M), hypertonic potassium chloride (KCl; 0.16 M), hypertonic (0.32 M) or hypotonic (0.16 M) mannitol, isotonic saline, or water delivered bilaterally via subcutaneous osmotic minipumps attached to intracranial cannulae. All cannulae terminated within the anterior hypothalamus-preoptic region. Hypertonic NaCl and KCl increased water intake over preinfusion levels in the majority of animals tested. However, the effects were variable, including some sizable increases as well as decreases. Hypertonic mannitol decreased daily water intake in 15 of 25 rats and produced essentially no change in the average intake of the group. Isotonic NaCl produced smaller increases and decreases, while water produced larger changes in individual rats, but neither solution had a significant effect on the average intake of the group. None of the infusates significantly altered food intake.