Doses Of Muscimol Research Articles

Nucleus Accumbens μ-Opioids Regulate Intake of a High-Fat Diet via Activation of a Distributed Brain Network

Endogenous opioid peptides within the nucleus accumbens, a forebrain site critical for the regulation of reward-related behavior, are believed to play an important role in the control of appetite. In particular, this system is thought to mediate the hedonic aspects of food intake, governing the positive emotional response to highly palatable food such as fat and sugar. Previous work has shown that intra-accumbens administration of the mu-opioid agonist D-Ala2,Nme-Phe4,Glyol5-enkephalin (DAMGO) markedly increases food intake and preferentially enhances the intake of palatable foods such as fat, sucrose, and salt. Using information from recently performed c-fos mapping experiments, we sought to explore the involvement of structures efferent to the nucleus accumbens in this feeding response. Free-feeding rats with dual sets of bilateral cannulas aimed at the nucleus accumbens and one of several output structures were infused with DAMGO (0, 0.25 microg/0.5 microl) in the accumbens, and fat intake was measured over a 2 hr period. Concurrent temporary inactivation with the GABA(A) agonist muscimol (5-20 ng/0.25 microl) of the dorsomedial hypothalamic nucleus, lateral hypothalamus, ventral tegmental area, or the intermediate region of the nucleus of the solitary tract blocked the robust increase in fat intake induced by intra-accumbens DAMGO at doses of muscimol that did not affect general motor activity. Muscimol alone also inhibited and augmented baseline fat intake in the lateral and dorsomedial hypothalamic nuclei, respectively. These results suggest that intake of energy-dense palatable food is controlled by activity in a neural network linking ventral striatal opioids with diencephalic and brainstem structures.

GABAergic control of food intake in the meat-type chickens

This study examined the effects of intracerebroventricular injections of γ-aminobutyric acid (GABA) agonists on short-term food intake in meat-type cockerels. In Experiment 1, birds were injected with various doses of muscimol, a GABA A agonist. In Experiment 2, the birds received bicuculline, a GABA A antagonist, prior to injection of muscimol. In Experiment 3, the effect of varying doses of baclofen, a GABA B agonist, on food intake was determined. The intracerebroventricular injection of muscimol caused a dose-dependent increase in food intake. This effect was significantly attenuated by pretreatment with bicuculline. Food intake was not affected by the intracerebroventricular injection of baclofen. These results suggest that GABA acts within the brain of broilers at a GABA A, but not GABA B, receptor to increase voluntary food intake.

The interaction of morphine and γ-aminobutyric acid (GABA)ergic systems in anxiolytic behavior: using μ-opioid receptor knockout mice

We investigated the interaction of morphine and γ-aminobutyric acid (GABA)ergic systems in anxiolytic action using μ-opioid receptor knockout and wild-type female mice. An elevated plus-maze test was used to assess anxiolytic behavior. The anxiolytic activities were monitored after experimental animals were pretreated with morphine [15 mg/kg, subcutaneous (s.c.)] and 3 h later received a single dose of muscimol (0.5 mg/kg, s.c.). In another experiment, mice received a single dose of opioid antagonist, naloxone [1.0 mg/kg, intraperitoneal (i.p.)], or GABAA receptor antagonist, (+)bicucullin (2.0 mg/kg, i.p.), 2.5 h after the morphine and 30 min before the muscimol injection. Control mice received vehicle only. The results show that morphine enhanced muscimol-induced staying time in open arms by 160% in wild-type mice. Moreover, the effect of morphine in the wild type was inhibited by the pretreatment of either naloxone or (+)bicucullin. Autoradiographic analysis indicated that morphine-administration raised [3H]muscimol binding by around 25% of the basal level in posterior thalamic, mediodorsal thalamic, and amygdaloid areas, but not in the hippocampal area in wild-type mice. In contrast, morphine administration did not alter the [3H]muscimol binding in μ-opioid receptor knockout mice. The present results reveal that μ-opioid receptor may play a role in the modification of anxiolytic behavior regulated by GABAergic neurotransmission.

The effects of ventral tegmental administration of GABA A, GABA B, NMDA and AMPA receptor agonists on ventral pallidum self-stimulation

The ventral pallidum (VP) is a basal forebrain structure that is interconnected with motor and limbic structures and may be considered as an interface between motivational and effector neural signals. Results from a considerable number of studies suggest that this structure is critically involved in reward-related behavior. The VP shares reciprocal connections with other reward-implicated regions, such as the ventral tegmental area (VTA). This anatomy predicts that drug-induced neuronal alterations in the VTA could profoundly alter the function of the VP. Here, using the curve-shift intracranial self-stimulation method, we studied the effects of muscimol (GABA A agonist), baclofen (GABA B agonist), NMDA and AMPA, microinjected bilaterally into the VTA on the rewarding efficacy of VP self-stimulation. Central injections of the highest dose of muscimol (0.128 μg) resulted in significant elevations in VP self-stimulation thresholds, indicating a reduction in the rewarding efficacy of the stimulation. Elevations in VP self-stimulation thresholds were also evident after intrategmental injections of higher doses of baclofen (0.12, 0.48 μg). By contrast, intrategmental activation of NMDA and AMPA receptors did not affect reward thresholds. These findings suggest that GABAergic and glutamatergic transmission in the VTA activate different circuits that may mediate different functions. Thus, the VTA–VP projection activated by GABA modulates VP stimulation reward, while the projection activated by glutamate may be involved in reward-unrelated effects, rather than in the processing of reward. The decreased rewarding efficacy of VP self-stimulation following intrategmental injections of muscimol and baclofen may be due to GABAergic modulation of ventral tegmental dopaminergic and nondopaminergic neurons projecting to the VP.

Effect of intracerebroventricular injection of GABA receptor agents on morphine-induced antinociception in the formalin test.

In the present study, the effects of gamma-aminobutyric acid (GABA) receptor agonists and antagonists on antinociception induced by morphine in the formalin test were investigated in rats. Intraperitoneal (i.p.) injection of different doses of morphine (1, 3, 6 and 9 mg/kg) and intracerebroventricular (i.c.v.) injection of different doses of muscimol (0.5, 1 and 2 microg per rat) or baclofen (0.25, 0.5 and 1 microg per rat) induced a dose-related antinociception in the both first and second phases of the formalin test. The responses induced by muscimol or baclofen in both phases were reduced by bicuculline or CGP35348 [p-(3-aminopropyl)-p-diethoxymethyl-phosphinic acid], respectively. Bicuculline alone has produced antinociception in the second phase and CGP35348 alone has had antinociception in both phases of the formalin test. Morphine in combination with different doses of muscimol or baclofen did not elicit potentiation. The opioid receptor antagonist naloxone reduced the response induced by muscimol in the second phase and baclofen in both phases of the formalin test. It may be concluded that central GABA(A) and GABA(B) receptor stimulation induces antinociception in the formalin test. However, the antinociception induced by GABA receptor agonists may be mediated partly through supraspinal opioid receptor mechanisms and, for the GABA(B) receptor agonist, through spinal and supraspinal opioid receptor mechanisms.

GABAA agents injected into the ventral pallidum differentially affect dopaminergic pivoting and cholinergic circling elicited from the shell of the nucleus accumbens

The ability of GABA(A) receptors in the ventral pallidum to modulate shell-specific behavior was studied. Injections of the non-selective acetylcholine receptor agonist, carbachol (5 microg), into the shell of the nucleus accumbens elicited contraversive circling, namely turning marked by normal stepping; in contrast, injections of a mixture of dopamine D(1) (SKF 38393, 5 microg) and D(2) (quinpirole, 10 microg) receptor agonists into this brain structure elicited contraversive pivoting, namely turning marked by abnormal hindlimb stepping. Unilateral injections of the GABA(A) receptor agonist muscimol (10, 25 and 50 ng) into the ventral pallidum dose-dependently mimicked shell-specific circling, especially when given at a level +8.6mm anterior to the interaural line; this effect was GABA(A) receptor specific, because it was prevented by the GABA(A) receptor antagonist bicuculline (150 ng). Unilateral pallidal injections of a dose of muscimol that was ineffective per se (10 ng) abolished contraversive pivoting elicited by shell injections of dopamine receptor agonists; instead, it elicited moderate ipsiversive pivoting. Pallidal injections of bicuculline (150 ng) replaced the contraversive pivoting elicited by dopamine receptor agonist with ipsiversive circling. In contrast, unilateral pallidal injections of 10 ng muscimol (anterior +8.6mm level) suppressed the contraversive circling elicited by shell injections of carbachol; instead, it elicited moderate ipsiversive pivoting. Pallidal injections of bicuculline (150 ng) produced short-lasting ipsiversive circling that was followed by contraversive pivoting. We conclude that the ventromedial portion of the ventral pallidum contains GABA(A) receptors that are crucial for the transmission of information from the shell of the nucleus accumbens via the ventral pallidum towards other brain structures; this holds especially for information about shell-specific circling elicited by carbachol. The same portion of the ventral pallidum also contains GABA(A) receptors that control the transfer of information from the nucleus accumbens towards structures outside the ventral pallidum; this holds especially for information about shell-specific pivoting elicited by dopaminergic agonists.

Involvement of central GABAergic neurons in basal and diurnal changes of tuberoinfundibular dopaminergic neuronal activity and prolactin secretion

Central administration of gamma-aminobutyric acid (GABA) has been shown to stimulate the secretion of prolactin (PRL). Whether GABA acts via dopamine, the major PRL-inhibiting hormone, and which GABA receptor type(s) is involved have not been ascertained. Both GABA A and GABA B receptor agonists and/or antagonists were administered centrally in this study and their effects on both basal and diurnal changes of tuberoinfundibular dopaminergic (TIDA) neuronal activity were determined by measuring the concentration of 3,4-dihydroxyphenylacetic acid (DOPAC) in the median eminence (ME). Serum PRL level was determined by RIA. Ovariectomized, estrogen-primed Sprague-Dawley rats implanted with intracerebroventricular (icv) cannulae were used. Muscimol (1 ng/3 μl/rat, icv), a GABA A receptor agonist, but not baclofen (1–100 ng/3 μl/rat, icv), a GABA B receptor agonist, injected in the morning significantly lowered and elevated ME DOPAC and serum PRL levels, respectively at 15 and 30 min. Lower and higher doses of muscimol were not effective. The effects of muscimol could also be prevented by co-administration of bicuculline (0.1–10 ng/3 μl, icv), a GABA A receptor antagonist. When bicuculline (10–500 ng/3 μl, icv) was given in the afternoon (at 1500 h), it significantly reversed the lowered ME DOPAC level in the afternoon and prevented the concurrent PRL surge. We conclude that endogenous GABA acting through GABA A receptors may play a significant role in the control of basal and diurnal changes of TIDA neuronal activity, and in turn, PRL secretion.

GABAergic modulation of descending inhibitory systems from the rostral ventromedial medulla (RVM). Dose-response analysis of nociception and neurological deficits

We have examined the effects of muscimol and bicuculline microinjected in the rostral ventromedial medulla (RVM) on motor function and on nociception in three pain tests. In Exp. 1 microinjection of muscimol (6.25–400 ng in 1 μl) in the RVM dose-dependently decreased pain threshold of rats and the ED 50 for muscimol was the same in both the hot plate and tail immersion pain tests. In the hot plate test, but not in the tail immersion test, paw withdrawal latencies increased again with high doses of muscimol (75–400 ng). High doses also produced catalepsy. Exp. 2 examined the effects of muscimol (50 ng) and bicuculline (50 ng) over a range of formalin concentrations (0.25–4%) in the formalin test. Muscimol increased responsiveness to formalin and reduced the slope of the formalin dose-response relation. Bicuculline decreased responses to formalin and reduced the slope of the formalin dose-response relation. It is suggested that RVM cells with inhibitory projections to the dorsal horn are not subject to strong GABAergic influence under mild noxious stimulation. RVM cells are thus active, and spinal dorsal horn relay neurons are inhibited. On the other hand, intense noxious peripheral stimulation may stimulate the release of GABA onto RVM cells, which in turn shuts off descending inhibitory fibers to allow transmission of nociceptor input through the dorsal horn.

Prenatal AZT or 3TC and mouse development of locomotor activity and hot-plate responding upon administration of the GABA(A) receptor agonist muscimol.

Zidovudine (AZT) and lamivudine (3TC) are nucleoside analogues administered prenatally in clinical practice, separately or in combination, as antiretroviral drugs to prevent HIV mother-to-child transmission by inhibiting viral reverse transcriptase. In animal studies pre- and/or perinatal exposure to AZT and 3TC induce age- and sex-dependent neurobehavioural alterations in the offspring. Investigation of short- and medium-term effects of in utero exposure to AZT or 3TC on development of the GABAergic system. Pregnant CD-1 mice were given orally twice daily AZT (160 mg/kg), 3TC (500 mg/kg) or vehicle solution (NaCl 0.9%) from pregnancy day 10 to delivery. Offspring locomotion and nociceptive sensitivity were examined on postnatal day (pnd) 8, 14, and 28 after administration of two doses of GABAergic agonist muscimol (pnd 8 and 14: 0.05 and 0.2 mg/kg; pnd 28: 0.2 and 1.0 mg/kg). A 30-min locomotor activity test and a 60 s hot-plate test (50+/-1 degrees C) were used. AZT and 3TC treated mice showed a mild increase of locomotor activity after administration of the high dose muscimol on pnd 8. On pnd 14 the low muscimol dose enhanced locomotor activity in vehicle and 3TC, but not in AZT pups, whereas no prenatal treatment effect was evident on pnd 28. AZT increased nociceptive sensitivity at all ages considered. Prenatal AZT effects on locomotor activity appear clearly detectable after GABAergic challenge and seem to be transient. AZT effects on pain sensitivity did not appear to be dependent on GABA regulated nociceptive mechanisms. Prenatal 3TC exposure had rather limited effects on locomotor activity development, and no effect on nociception.

GABAergic contribution to rat bladder hyperactivity after middle cerebral artery occlusion.

To evaluate the influences of gamma-aminobutyric acid (GABA) mechanisms on bladder hyperactivity after left middle cerebral artery occlusion, cystometric recordings were obtained from unanesthetized female rats. Intracerebroventricular administration of both muscimol (GABA(A) receptor agonist; 0.1-10 nmol) and baclofen (GABA(B) receptor agonist; 0.1-3 nmol) produced dose-dependent inhibitions of micturition with increases in bladder capacity (BC). The effects of high doses (1-10 nmol) were similar in sham-operated (SO) and cerebral-infarcted (CI) rats. However, lower doses of muscimol (0.1 or 0.3 nmol) and baclofen (0.1 nmol) reduced BC in CI rats. After bicuculline (GABA(A) receptor antagonist; 1 or 3 nmol) administration, BC in both SO and CI rats first decreased and subsequently increased. An increase in urethral pressure was observed after administration of bicuculline (3 nmol) but not with either muscimol or baclofen. Infarct volumes in muscimol-, bicuculline-, or baclofen-treated rats were not significantly different from those of vehicle-treated rats. These results suggest that GABAergic mechanisms inhibit the micturition reflex at the supraspinal level but that this can change as a result of CI.

Postural and Anticonvulsant Effects of Inhibition of the Rat Subthalamic Nucleus

The subthalamic nucleus (STN) plays a crucial role as a regulator of basal ganglia outflow by providing excitatory glutamatergic input into the two output nuclei of the basal ganglia, substantia nigra pars reticulata (SNpr), and entopeduncular nucleus. This study examined the effects of suppressing activity in the STN of the awake, behaving rat. Specifically, we evaluated the effects of unilateral and bilateral focal inhibition of STN on posture, locomotion, and susceptibility to limbic motor seizures. Unilateral microinjection of a GABA(A) receptor agonist (muscimol, 200 pmol) into STN produced a site-dependent contralaterally directed postural asymmetry without locomotor activation. This effect differed from responses produced by the same dose of muscimol placed into SNpr, which included locomotor activation in addition to contralaterally directed postural asymmetry. Locomotor activation and postural asymmetry were obtained also after blockade of glutamate transmission in SNpr by the unilateral application of kynurenate (100 nmol). Our observation that STN inhibition did not induce the locomotor activation characteristic of SNpr inhibition suggests that there are glutamatergic inputs to SNpr, other than those from STN, that are responsible for controlling locomotion. Bilateral, but not unilateral, injection of muscimol (200 pmol) into STN protected against limbic motor seizures evoked either by intravenous bicuculline or by focal application of bicuculline into anterior piriform cortex (area tempestas). These results demonstrate that focal inhibition of STN reproduces the postural asymmetry and anticonvulsant actions that are obtained with the inhibition of SNpr. This provides behavioral support for the concept that STN contributes a crucial tonic excitatory (glutamatergic) drive to the rat SNpr.

Intrastriatal administration of 5-aminolevulinic acid induces convulsions and body asymmetry through glutamatergic mechanisms

The involvement of glutamatergic and GABAergic mechanisms in the behavioral effects induced by the intrastriatal injection of 5-aminolevulinic acid (ALA) (1–8 μmol/2 μl), a metabolite that accumulates in porphyrias, was evaluated. ALA administration to adult female rats increased locomotor activity, induced clonic convulsions and elicited dose-dependent body asymmetry assessed by the elevated body swing test. ALA-induced convulsions were prevented by intrastriatal preadministration of the glutamate antagonists, 6,7-dinitroquinoxaline-2,3-dione (8 nmol/0.5 μl) or dizocilpine (2.5 nmol/0.5 μl), but not by the GABA agonist, muscimol (46 pmol/0.5 μl). Body asymmetry was prevented only by 6,7-dinitroquinoxaline-2,3-dione pretreatment. A higher dose of muscimol (92 pmol/0.5 μl) prevented both ALA-induced convulsions and body asymmetry. However, this dose of muscimol induced motor biases, which make difficult to ascertain the involvement of GABA A receptors in ALA-induced behavioral effects. This study suggests that glutamatergic mechanisms underlie the ALA-induced convulsions and body asymmetry. The present results may be of value in understanding the physiopathology of the neurological dysfunction occurring in acute porphyrias.

Brain allopregnanolone regulates the potency of the GABA A receptor agonist muscimol

Allopregnanolone (ALLO), a potent positive-allosteric modulator of the action of GABA at GABA A receptors, is synthesized in the brain from progesterone by the sequential action of two enzymes: 5α-reductase and 3 α-hydroxysteroidoxidoreductase. The concentration of ALLO in various parts of the mouse brain varies substantially, from 15 pmol/g in the olfactory bulb, to approximately 6 pmol/g in the frontoparietal cortex, and 2.7 pmol/g in the cerebellum. The systemic administration of 48 μmol/kg of the Type I and Type II 5α-reductase inhibitor, (17β)-17-[bis (1-methylethyl) amino carbonyl)] androsta-3, 5-diene-3-carboxylic acid (SKF 105,111), reduced brain ALLO content by 80–90% in 30 min; the rate constant ( k) of ALLO decrease in each brain area can be utilized to establish the rate of ALLO biosynthesis, which is higher in the olfactory bulb (62 pmol/g/h) than in the frontoparietal cortex (24 pmol/g/h) or cerebellum (11 pmol/g/h). The duration of the righting reflex loss elicited by the potent GABA A receptor agonist muscimol was reduced in SKF 105,111-treated ALLO-depleted mice. SKF 105,111 treatment had no effect on muscimol metabolism or on brain levels of pregnenolone and progesterone; however, the brain levels of 5α-DHP, the precursor of ALLO, were also decreased. Administration of ALLO at a dose of 15 μmol/kg ip by itself did not alter the muscimol-induced loss of the righting reflex; but it completely blocked the effect of SKF 105,111. To elucidate the possible molecular mechanism by which a decrease of brain ALLO content can shorten the duration of the righting reflex loss elicited by muscimol, we patch-clamped neocortical pyramidal neurons of mice pretreated with SKF 105,111 or vehicle, and studied the efficiency of muscimol in eliciting Cl − currents. The current amplitude was significantly smaller in neurons from SKF 105,111-treated mice, especially at lower doses (0.1–1 μM) of muscimol, and the muscimol dose–response (0.1–10 μM) relationship displayed cooperativity ( n H=1.4). These data suggest that ALLO synthesized in brain plays an important physiological permissive role in the modulation of GABA-gated Cl − channel function.

A comparison of the effects of bilateral and unilateral infusions of muscimol into the basal forebrain on cued detection of visual targets in rats.

This study investigated the role of the basal forebrain cholinergic system (BFCS) in rats' performance of a visuospatial attention task. Muscimol was infused bilaterally and unilaterally into the BFCS to inhibit cholinergic projections to the cortex. Muscimol slowed responding without significantly affecting side-bias. Bilateral infusions increased accuracy for all targets, whereas unilateral infusions reduced accuracy for targets contralateral to the infusion and increased accuracy for targets ipsilateral to the infusion. After a low unilateral dose of muscimol, invalid cues impaired detection of contralateral targets and spared detection of ipsilateral targets. A high unilateral dose of muscimol impaired detection of contralateral targets independently of cueing. These results suggest that interhemispheric imbalance in cortical activity by pharmacological manipulation of the BFCS can impair the detection of lateralized visual stimuli.

Differential modulation by baclofen on antinociception induced by morphine and β -endorphin administered intracerebroventricularly in the formalin test

Our previous studies have demonstrated that supraspinal GABAergic receptors are differentially involved in the antinociception induced by morphine and β-endorphin given intracerebroventricularly (i.c.v.) in the tail-flick and hot-plate tests. These two models employed a phasic, thermal nociceptive stimulus. The present study was designed to examine the possible involvement of supraspinal GABAergic receptors in opioid-induced antinociception in the formalin test. Morphine (1 μg) and β-endorphin (1 μg) given i.c.v. displayed the almost complete inhibitory effects against the hyperalgesic response in both phases. Muscimol (75–100 ng) and baclofen (5–10 ng) injected i.c.v. produced the hypoalgesic response in the both phases. The hypoalgesic response induced by muscimol and baclofen observed during the second phase was more pronounced than that observed during the second phase. Baclofen (2.5 ng), at the dose which did not affect the hyperalgesic response, resulted in a significant reversal of the i.c.v. administered β-endorphin-induced hypoalgesic response observed during the second, but not the first, phase. However, the hypoalgesic response induced by i.c.v. administered morphine was not changed by the same dose of muscimol or baclofen injected i.c.v. Our results indicate that, at the supraspinal level, GABABreceptors appear to be involved in the modulation of antinociception induced by supraspinally administered β-endorphin, but not morphine, in the formalin test model.

Neurochemical modulation of the P13 midlatency auditory evoked potential in the rat

Previous studies have shown that the vertex-recorded P13 auditory evoked potential in the rat appears to be the rodent equivalent of the human P1 (or P50) potential. This sleep state-dependent potential appears to be generated, at least in part, by cholinergic pedunculopontine nucleus projections. The present studies used localized microinjections of neuroactive compounds into the region of the pedunculopontine nucleus in order to modulate the vertex-recorded P13 potential. Both the GABAergic agonist, muscimol, and the noradrenergic α 2 receptor agonist, clonidine, were found to reduce the amplitude of the P13 potential in a dose-dependent manner. The suppressive effect of clonidine on P13 potential amplitude was blocked by pretreatment with the noradrenergic α 2 receptor antagonist, yohimbine. In addition, habituation of the P13 potential, measured using a paired stimulus paradigm, was increased by microinjection of a dose of muscimol or clonidine which did not change the amplitude of the P13 potential induced by the first stimulus of a pair. In contrast, microinjection of yohimbine decreased habituation of the P13 potential. These results show that the vertex-recorded P13 potential and its habituation can be modulated by activation of known inhibitory synapses, both GABAergic and noradrenergic, at the level of the pedunculopontine nucleus. This provides further evidence that the P13 potential is generated, at least in part, by pedunculopontine nucleus outputs.

Activation of GABAA but not GABAB receptors in the NTSblocked bradycardia of chemoreflex in awake rats.

In the present study we analyzed effects of bilateral microinjections of muscimol (a GABAA agonist) and baclofen (a GABAB agonist) into the nucleus tractus solitarius (NTS) on bradycardic and pressor responses to chemoreflex activation (potassium cyanide, 40 micrograms/rat iv) in awake rats. Bilateral microinjections of muscimol (25 and 50 pmol/50 nl) into the NTS increased baseline mean arterial pressure (MAP): 119 +/- 8 vs. 107 +/- 2 mmHg (n = 6) and 121 +/- 8 vs. 103 +/- 3 mmHg (n = 6), respectively. Muscimol at 25 pmol/50 nl reduced the bradycardic response to chemoreflex activation 5 min after microinjection; with 50 pmol/50 nl the bradycardic response to chemoreflex activation was reduced 5, 15, 30, and 60 min after microinjection. Neither muscimol dose produced an effect on the pressor response of the chemoreflex. Effects of muscimol (50 pmol/50 nl) on basal MAP and on the bradycardic response of the chemoreflex were prevented by prior microinjection of bicuculline (a GABAA antagonist, 40 pmol/50 nl) into the NTS. Bilateral microinjections of baclofen (12.5 and 25 pmol/50 nl) into the NTS produced an increase in baseline MAP [137 +/- 9 vs. 108 +/- 4 (n = 7) and 145 +/- 5 vs. 105 +/- 2 mmHg (n = 7), respectively], no changes in basal heart rate, and no effects on the bradycardic response; 25 pmol/50 nl only attenuated the pressor response to chemoreflex activation. The data show that activation of GABAA receptors in the NTS produces a significant reduction in the bradycardic response, whereas activation of GABAB receptors produces a significant reduction in the pressor response of the chemoreflex. We conclude that 1) GABAA but not GABAB plays an inhibitory role in neurons of the lateral commissural NTS involved in the parasympathetic component of the chemoreflex and 2) attenuation of the pressor response of the chemoreflex by activation of GABAB receptors may be due to inhibition of sympathoexcitatory neurons in the NTS or may be secondary to the large increase in baseline MAP produced by baclofen.

Blocking GABA A receptors in the anterior ventral tegmental area attenuates ethanol intake of the alcohol-preferring P rat

The effect of blocking the A subtype of gamma-aminobutyric acid (GABA(A)) receptors in the anterior ventral tegmental area (VTA) on ethanol (EtOH; 10% v/v) and saccharin (SACC; 0.0125%) consumption was investigated in alcohol-preferring P rats. Picrotoxin (0.005, 0.01, 0.05 and 0.10 microg/0.5 microl) was injected into the VTA, and consumption of EtOH and SACC was assessed in two 2-h limited-access drinking paradigms (concurrent EtOH/SACC access, and alternate-day-access to EtOH and SACC). Under concurrent-access conditions, the picrotoxin microinjections resulted in a 55 and 84% decrease in EtOH consumption at the 0.05 and 0.10 microg doses, respectively, compared with consumption following microinjections of vehicle solution (P<0.05). Saccharin intake was not significantly altered by picrotoxin. Under alternate-day-access drinking conditions, the picrotoxin microinjections resulted in dose-dependent decreases in EtOH consumption of 37-68%, with significant decreases following the 0.005, 0.05 and 0.10 microg doses (P<0.04). Saccharin intake was significantly reduced only at the 0.05 microg dose. The decrease in EtOH consumption after 0.10 microg picrotoxin was attenuated by co-administration of 0.01 microg muscimol. This dose of muscimol had no effect on EtOH consumption when injected alone. Intra-VTA injections of bicuculline (0.04 microg), another GABA(A) antagonist, reduced EtOH intake, comparable to the reduction following 0.10 microg picrotoxin. Microinjections of 0.10 microg picrotoxin in regions outside the VTA failed to decrease EtOH intake. These results suggest that anterior VTA mechanisms regulating alcohol drinking behavior are under tonic GABA inhibition, mediated by GABA(A) receptors. The results also suggest that different neural mechanisms are regulating voluntary EtOH and SACC drinking behaviors.

INTERACTION OF ANTINOCICEPTIVE EFFECTS OF MORPHINE AND GABA AGONISTS AT THE LEVEL OF SPINAL CORD IN RATS

S454 Pharmacological and neurophysical data suggest an antinociceptive interaction between GABA and opioid receptors. This is supported by the potentiation of GABA-induced current by opioid [1], the enhancement of opioid antinociception by benzodiazepines [2], which mainly act on GABAA receptor, and the spinal antinociception of GABA agonists. In contrast, intracerebroventricular injection of GABAA agonist inhibits the morphine antinociceptive effect [3]. However, there have been little information regarding their interaction in the spinal cord. In the present study, we have examined the effects of GABAA agonist, muscimol and GABAB agonist, baclofen on the both of somatic and visceral antinociceptive effects of morphine at the level of spinal cord. METHODS: Following Animal Care approval, an intrathecal catheter was implanted from L4/L5 level in male Sprague Dawley rats. Tail flick (TF) test and colorectal distension (CD) test were employed to measure response to somatic stimulus and visceral stimulus, respectively. In the CD test, the pressure at which abdominal contractions were triggered was defined as the threshold when pressure within the intracolonic balloon was steadily increased. Measurements were performed before and 5, 10, 15, 20, 30, 60, 90, 120 and 180 min after intrathecal administration of morphine, muscimol, baclofen, the combination of those (morphine 1 or 0.1 [micro sign]g and muscimol 0.2 [micro sign]g or baclofen 0.03 [micro sign]g) or saline. Percent of maximum possible effect (%MPE) was calculated by transforming response threshold in TF and CD tests. RESULTS: Intrathecally administered morphine, muscimol or baclofen produced the both somatic and visceral antinociception in a dose dependent fashion. Morphine 1 [micro sign]g increased mean %MPE for 90 min (with a peak effect of 96%MPE at 20 min) in TF test and for 30 min (with a peak effect of 53%MPE at 20 min) in CD test. The combination of 1 [micro sign]g morphine and an ineffective or slightly effective dose of muscimol (0.2 [micro sign]g) or baclofen (0.03 [micro sign]g) produced significant increases in %MPEs compared with morphine 1 [micro sign]g in both tests. Morphine 0.1 [micro sign]g showed slightly increased %MPE in TF test but not in CD test. It was potentiated by GABA agonists in TF test but not in CD test. CONCLUSIONS: The results demonstrate that GABAA, GABAB agonist may potentiate the both somatic and visceral antinociceptive effects of opioid at the level of spinal cord.

Muscimol and midazolam do not potentiate each other's effects on sleep EEG in the rat.

The interaction of a gamma-aminobutyric acid-A (GABAA) receptor agonist and a benzodiazepine-type modulator of GABAA receptors on sleep was investigated. Low doses of muscimol (0.3 mg/kg) and the benzodiazepine midazolam (1.5 mg/kg) were administered alone and in combination, in random order, to eight rats. All injections were given intraperitoneally at light onset. Electroencephalogram (EEG) and electromyogram were recorded during the first 6 h post injection. Compared with vehicle, muscimol hardly affected the time spent in non-rapid eye movement sleep (non-REMS) and REMS, but significantly enhanced EEG activity in the frequency range between 2 and 6 Hz during non-REMS. Midazolam significantly increased the time spent in non-REMS, reduced EEG activity at frequencies < 12 Hz, and elevated EEG activity in most higher frequencies during this state. The combined administration of muscimol and midazolam affected non-REMS-specific EEG activity in an unexpected fashion: the effects were intermediate between those of muscimol and midazolam. These results indicate that muscimol and midazolam have dissimilar effects on EEG within non-REMS and demonstrate that midazolam does not augment but attenuates the muscimol-induced changes in sleep EEG. Our data are at variance with established mechanisms, according to which agonistic modulators would have similar effects and should potentiate the effects of GABAA agonists. The present data suggest that application of agonists and agonistic modulators of GABAA receptors causes differential net effects on sleep parameters.