Search

Overconsumption, or eating beyond the point of homeostasis, is a key feature in the development of obesity. Although people are consuming beyond the point of homeostasis, they are not consuming constantly or indefinitely. Thus, there is likely a mechanism that acts to terminate periods of food intake at some point beyond satiation and prior to aversion, or the negative effects of extreme excess (nausea, bloating, etc.). The purpose of the present study was to assess the lateral habenula as a candidate region for such a mechanism, due to its connectivity to midbrain reward circuitry, sensitivity to metabolic signaling, and pronounced role in drug-related motivated behaviors. Two groups of male Sprague-Dawley rats were surgically implanted with bilateral guide cannula targeting the LHb. Rats were then habituated to feeding chambers, wherein locomotion and food intake were monitored throughout a two-hour session. One experimental group was tested in the presence of rat chow; the second group was instead given access to a sweetened fat diet. Each subject separately received a 0.2 μL vehicle (0.9% saline solution) and baclofen-muscimol (50 ng/0.2 μL of each drug dissolved in 0.9% saline) injection. Additionally, on a third injection day, each rat received an injection of mu-opioid agonist DAMGO (0.1 μg/0.2 μL) prior to placement in the chamber. LHb inactivation did not result in significant alterations in feeding behavior, but produced a consistent increase in locomotor activity in both experimental groups. Mu-opioid receptor stimulation increased feeding on standard chow, but decreased intake of the sweetened-fat diet. Although LHb inactivation did not increase feeding as predicted, the novel finding that mu opioid receptor stimulation decreased feeding on a highly palatable diet, but increased intake of rat chow, highlights a differential role for the LHb in regulating hedonic consummatory behavior.

Made available in DSpace on 2020-12-12T02:40:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-07-13 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Injection of muscimol, a GABAA receptor agonist, into the lateral parabrachial nucleus (LPBN) induces 0.3 M NaCl intake in rats. In the present work, we investigated whether such an effect applies to hypertonic (0.3 M) mineral solutions in general or is selective to sodium solutions in a 240 min intake test. Muscimol injection (0.5 nmol/0.2 μL) compared to vehicle injection into the LPBN of adult hydrated rats produced a preferential ingestion of 0.3 M NaCl (25.3 ± 10.2 mL) followed by a 0.3 M NaHCO3 intake (11.7 ± 5.6 mL), with no significant effect on water, KCl and CaCl2 intake. Only the effect of muscimol on NaCl intake (19.0 ± 10.4 mL) persisted in cell-dehydrated rats, with hardly any effect on water or other mineral solutions. The results suggest that the LPBN controls the ingestion of hypertonic NaCl and NaHCO3. They also suggest a selective mechanisms involving the LPBN to check hypertonic sodium intake. Department of Basic Sciences School of Dentistry Sao Paulo State University (UNESP), Rodovia Marechal Rondom, Km 527 Department of Physiology and Pathology School of Dentistry UNESP Department of Basic Sciences School of Dentistry Sao Paulo State University (UNESP), Rodovia Marechal Rondom, Km 527 Department of Physiology and Pathology School of Dentistry UNESP

Objective Suppression of the gamma-aminobutyric acid (GABA)ergic activity of the zona incerta (ZI) reportedly plays a role in neuropathic pain after spinal cord injury (SCI). A reduction in GABAergic signaling in the ZI of a thoracic hemisection-SCI rat model has been suggested, but not clearly demonstrated. Accordingly, our objective was to investigate whether GABAergic signals influence SCI-induced neuropathic pain. Methods In vivo, we recorded and compared single-unit, neuronal activity between hemisection-SCI and sham-operated rat models. Furthermore, we analyzed neuronal activity in both models following treatment with either a GABAA receptor agonist (muscimol) or antagonist (bicuculline). Results Rats that underwent hemisection SCI exhibited reduced hindpaw withdrawal thresholds, latencies, and decreased ZI neuronal activity compared with sham-operated controls. Importantly, muscimol treatment increased, whereas bicuculline decreased, the firing rates of the ZI neurons. The muscimol treated, hemisection-SCI rats also exhibited increased hindpaw withdrawal thresholds and latencies. Conclusions These data provide evidence that neuropathic pain after SCI is caused by decreased GABAergic signaling in the ZI. Furthermore, our data demonstrate that infusion of a GABAergic drug into the ZI could restore its inhibitory action and improve neuropathic pain behaviors.

a b s t r a c t Abnormal enhancement of polysynaptic brainstem reflexes has been previously reported in patients with spinal cord injury (SCI). We aimed to investigate trigemino-cervical reflex (TCR) in SCI since it may reflect alterations in the connections of trigeminal proprioceptive system and cervical motoneurons. Consecutive 14 patients with SCI and 16 healthy subjects were included in this study. All patients were in the chronic phase. TCR was recorded over sternocleidomastoid (SCM) and splenius capitis (SC) muscles by stimulation of infraorbital nerve. We measured onset latency, amplitudes and durations of responses and compared between groups. We obtained stable responses over both muscles after one sided stimulation in healthy volunteers whereas probability of TCR was decreased in patients over both SCM (78.6% vs. 100%, p = 0.050) and SC (71.4% vs. 100%, p = 0.022). The absence of TCR was related to use of oral baclofen (≥50 mg/day). However, when present, responses of SCI group had higher amplitudes and were more persistent. We demonstrated that TCR probability was similar to healthy subjects in SCI patients who used no or low dose oral baclofen. But it had higher amplitudes and longer durations. It was not obtained in only two patients who used oral baclofen more than 50 mg/day.

Ischemia is one of the most prominent risk factors of neurodegenerative diseases such as Alzheimer's disease. The effects of oxygen and glucose depletion in hippocampal tissue due to ischemia can be mimicked in vitro using the oxygen and glucose deprivation (OGD) model. In this study, we applied OGD on acute rat hippocampal slices in order to design an elementary yet quantitative histological technique that compares the neuroprotective effects of (l)-carnitine to known neuroprotectors, such as the N-methyl-d-aspartate (NMDA) receptor antagonist memantine and the gamma-aminobutyric acid (GABA)-B receptor agonist baclofen. The level of neurodegeneration and the efficiency of pharmacological applications were estimated via stratum pyramidale width measurements in CA1 and CA3 regions of Nissl-stained 200-μm thick hippocampal slices. We demonstrated that (l)-carnitine is an effective pharmacological target against the neurodegeneration induced by in vitro ischemia in a narrow range of concentrations. Even though the effect of chemical neuroprotection was significant, full recovery was not achieved in the dose interval of 5-100μM. In addition to chemical applications, hypothermia was used as a physical neuroprotection against ischemia-related neurodegeneration. Our results showed that incubation of slices for 60min at 4°C provided the same level of neuroprotection as the most effective doses of memantine, baclofen, and (l)-carnitine.

While our previous work suggests that the midazolam-induced memory impairment results from the inhibition of new association formation, little is known about the neural correlates underlying these effects beyond the effects of GABA agonists on the brain. We used arterial spin-labeling perfusion MRI to measure cerebral blood flow changes associated with the effects of midazolam on ability to learn arbitrary word-pairs. Using a double-blind, within-subject cross-over design, subjects studied word-pairs for a later cued-recall test while they were scanned. Lists of different word-pairs were studied both before and after an injection of either saline or midazolam. As expected, recall was severely impaired under midazolam. The contrast of MRI signal before and after midazolam administration revealed a decrease in CBF in the left dorsolateral prefrontal cortex (DLPFC), left cingulate gyrus and left posterior cingulate gyrus/precuneus. These effects were observed even after controlling for any effect of injection. A strong correlation between the midazolam-induced changes in neural activity and memory performance was found in the left DLPFC. These findings provide converging evidence that this region plays a critical role in the formation of new associations and that low functioning of this region is associated with anterograde amnesia.

In the present study, functional roles of GABA receptors in the nucleus accumbens on morphine self-administration behavior were investigated. Male Sprague-Dawley rats were trained to press lever for morphine (0.1 mg/kg per infusion) during daily 1-h self-administration session. After establishing stable baseline responses, rats were given microinjections of the GABA(A) receptor agonist muscimol (0, 250 and 500 ng/microl, bilateral) or the GABA(B) receptor agonist baclofen (0, 100 and 250 ng/microl, bilateral) into the nucleus accumbens immediately before the morphine self-administration. Microinjection of muscimol (250 and 500 ng/microl) into the nucleus accumbens, but not baclofen, decreased morphine self-administration responses. These results suggest that activation of GABA(A) receptors, but not GABA(B) receptors, in the nucleus accumbens plays a critical role in modulating the reinforcing effects of morphine.

The cerebellum has been proved to be essential for the acquisition of delay eyeblink conditioning, but its contribution to the acquisition of trace eyeblink conditioning (TEBC) has not been fully determined. In the present study, using chemically reversible inactivation techniques, we examined the relative contribution of ipsilateral cerebellum to the acquisition of TEBC using different time length of trace interval (TI) in guinea pigs. It was found that inactivations of the left intermediate cerebellum with a GABA(A) receptor agonist muscimol during training completely prevented the acquisition of TEBC using a relatively short (50 ms) TI, instead of the acquisition of TEBC using a relatively long (250 ms) TI. However, inactivations of the left intermediate cerebellum totally abolished the well-established left trace conditioned eyeblink responses (CRs) regardless of the time length of TI. These results suggested that while the ipsilateral cerebellum is essential for the expression of trace CRs, its contribution to the acquisition of trace CRs appears to mainly depend on the time length of TI.

The purpose of this study was to investigate the role of the ventromedial prefrontal cortex (vmPFC) in the expression of extinguished heroin seeking as measured by conditioned place preference (CPP) in males Sprague-Dawley rats (n=25). Heroin place conditioning (0.3mg/kg SC x 4 sessions) was followed by a test of preference 24h later, extinction (saline x 4 sessions), heroin reconditioning (saline or 1.0mg/kg x 1 session), and a second test of place preference 24h later. Fifteen minutes prior to this test, rats received intra-vmPFC infusions (bilateral, 0.5 microl/side) of a mixture of GABA(A) (muscimol; 0.03 nmol) and a GABA(B) (baclofen; 0.3 nmol) agonists, or vehicle. As expected on the basis of previous studies, reconditioning with heroin resulted in the re-emergence of a CPP. Importantly, inactivation of the vmPFC produced the same effect in animals that did not receive heroin on the session of reconditioning. These results indicate that the vmPFC modulates expression of extinguished heroin seeking and suggest that prefrontal inhibitory mechanisms are involved in relapse to drug seeking.

Cocaine-primed reinstatement of drug-seeking is associated with a decrease in extracellular GABA in the ventral pallidum (VP). The present study investigated the neural mechanism of this cocaine-induced decrease in VP GABA by determining if activity of the glutamatergic projection from the medial prefrontal cortex (PFC) to the nucleus accumbens is required for the effect. Microdialysis was performed to measure extracellular GABA in the VP while simultaneously, either a combination of the GABA agonists baclofen and muscimol was microinjected into the PFC, or the AMPA/kainate glutamate receptor antagonist CNQX was microinjected into the accumbens core. Inhibition of the PFC with GABA agonists and blockade of AMPA glutamate receptors in the accumbens core were both sufficient to prevent the cocaine-induced decrease in VP GABA, further implicating increased activity of the cortico-striato-pallidal circuit in relapse to drug-seeking.

Changes in the activity of putative interneurons of the stratum oriens of the hippocampus and hippocampal EEG after the delivery of the GABA A receptor antagonist bicuculline (1.0 nmol/1 μl) to the medial septal region were investigated in awake rabbits. The injection of bicuculline produced a sharp increase in the firing rate in 94.3% of hippocampal cells. The effect of bicuculline on the neuronal theta rhythmicity depended on the number of injections. The first five daily infusions decreased the theta activity in 85.7% of cells. On the fourth to fifth experimental days paroxysmal discharges and 8–15 Hz oscillations were recorded in the hippocampal EEG. Six to seven further daily bicuculline injections following a brief diminution of theta activity produced a sharp augmentation of theta oscillations in 78.9% of cells and provoked seizures. Immediately before seizures, stabilization of theta bursts and an increase in burst frequency was usually observed in putative interneurons. During seizures, neuronal rhythmic activity was either disordered and then turned into seizure discharges or was inhibited, partially or completely. In the hippocampal EEG, the power of theta rhythm before seizures usually strongly increased compared with controls. Injection of the GABA A agonist muscimol (30 nmol/1 μl) 15 min before bicuculline infusion prevented the development of seizures. These findings suggest that the interplay between septal neurons via GABA A receptors is critical in the tuning of septal output signals that insure generation of natural theta rhythm as well as adequate functioning of the hippocampus.

It is well established that the reinforcing effect of drugs of abuse is linked to the mesolimbic dopamine (DA) system. Morphine produces an increase in DA release in the brain, which may provide positive reinforcement contributing to the development of motivational aspects of drug-seeking and maintenance behavior. Several studies suggest that the GABA receptor system may play a significant role in the modulating the mesolimbic DA system. The purpose of this study was to investigate potential roles for GABA agonists in morphine self-administration behavior. Male Sprague-Dawley rats were trained to self-administrated morphine (0.1 mg/kg per injection) during daily 1-h sessions under a fixed ratio 1 schedule. Rats received an intravenous injection of the selective GABA(B) antagonist SCH 50911 (2.0 mg/kg) or an intraperitoneal injection of the GABA(A) antagonist bicuculline (1.0 mg/kg), immediately followed by either an intraperitoneal injection of baclofen (1.25 or 1.8 mg/kg) or muscimol (0.5 or 1.0 mg/kg, i.p.), 30 min prior to the start of test session. Results showed that pretreatment with baclofen or muscimol reduced morphine-maintenance response in a dose-dependent fashion and that baclofen and muscimol effects were reversed by injections of SCH 50911 and bicuculline, respectively. These data suggest that activation of both GABA(A) and GABA(B) receptors may be effective in suppressing the reinforcing effects of morphine.

The mechanisms underlying gamma-amino butyric acid (GABA(B)) receptor-mediated inhibition of exocytosis have been characterized in a variety of synapses. Using patch-clamp recording methods, we attempted to clarify the intracellular mechanisms underlying presynaptic inhibition in autaptic synapses of isolated mouse hippocampal neurons. Baclofen, a selective GABA(B) receptor agonist, decreased the frequency of glutamatergic miniature excitatory postsynaptic currents (mEPSCs) without changing their amplitude in Ca(2+)-free extracellular solution, suggesting that baclofen inhibits exocytosis downstream of Ca(2+) entry. Syntaxin 1A is known to modulate exocytosis and suppress neuronal sprouting. Antisense oligonucleotide-mediated knockdown of syntaxin 1A increased the frequency of mEPSCs under Ca(2+)-free condition. Estimation of the number of functional release sites by staining with FM1-43 indicated that the increased frequency of mEPSCs was induced by facilitation of exocytosis at each site, rather than by an increased number of release sites due to neuronal sprouting. Baclofen reduced mEPSC frequency in syntaxin 1A-knockdown neurons to the same level as that in nonsense oligonucleotide transfected neurons under Ca(2+)-free condition. These results suggest that the GABA(B) receptor- and syntaxin 1A-induced inhibitions of exocytosis occlude one another and that the GABA(B) receptor shares a common intracellular pathway with syntaxin 1A in inhibiting transmitter release downstream of Ca(2+) entry.

The preoptic area (POA) is thought to play an important role in thermoregulation and fever. Local application of prostaglandin E2 (PGE2) to this region elicits increases in core body temperature, heart rate, and plasma levels of adrenocorticotrophic hormone (ACTH). Similar effects on body temperature and heart rate have also been reported after local application of the GABA A receptor agonist muscimol to the preoptic area. The purpose of this study was to assess and compare the effects of microinjection of PGE2 and muscimol into the preoptic area in the same chronically instrumented conscious rats on plasma levels of ACTH. Injection of either PGE2 (150 pmol/100 nL) or muscimol (20 or 80 pmol/100 nL) into the same sites in the preoptic area evoked increases in body temperature, heart rate, blood pressure, and plasma levels of ACTH, while significant increases in locomotor activity were apparent only after muscimol. These data confirm and extend previous findings and support the notion that neurons in the region of the preoptic area exert tonic inhibition on downstream mechanisms capable of increasing the activity of the hypothalamic–pituitary–adrenal (HPA) axis as well as sympathetic thermogenic and cardiac activity.

GABA(B) agonists inhibit excitatory transmission to hippocampal CA3 neurons during low frequency stimulation. We examined whether GABA(B) receptor activation can also enhance synaptic efficacy, when investigated at an input with high initial release probability. Short-term depression of field excitatory postsynaptic potential (EPSP) amplitude was observed during trains of stimuli applied to associational/commissural inputs (10-50 Hz; 22 degrees C). Baclofen (10 microM) reduced the amplitude of initial EPSPs in a train, and also reduced the degree of short-term depression. EPSPs recorded late in a train were significantly larger in baclofen than those recorded in control solution. These dual effects were mimicked by another selective GABA(B) agonist (SKF 97541, 10 microM), and abolished by a GABA(B)-selective antagonist (SCH 50911, 20 microM). The effects of baclofen were similar at a higher recording temperature (32 degrees C), where short-term depression was observed at higher stimulation frequencies. These results are consistent with the idea that a reduction of transmitter release probability could increase the fidelity of high-frequency transmission at this input, an effect that could help account for excitatory effects of GABA(B) agonists in some seizure models.

An in vitro mouse hemisected spinal cord was used to characterize the γ-aminobutyric acid receptor type B (GABA B ) modulation of the ventral root potential (VRP) in response to electrical stimulation of the dorsal root (DR). Low-intensity (LI) and high-intensity (HI) stimulation induced VRPs with progressively higher amplitude and duration. Repetitive HI-stimulation of the DR (1–10 Hz) produced windup. The selective GABA B receptor agonist, CGP35024, inhibited the VRPs in a dose-dependent manner. The inhibitory action of CGP35024 was blocked by CGP52432, a potent GABA B receptor antagonist. Following washout of the GABA B receptor agonist, VRPs and windup were significantly enhanced. The rebound increase of the VRP following removal of CGP35024 was also blocked by the GABA B receptor antagonist, CGP52432. This phenomenon is not linked to receptor desensitization, but rather due to GABA B receptor-induced hyperactivity of N-, P/Q-type Ca 2+ channels, as ω-CgTx GVIA and MVIIC abolished/prevented the increase. The ‘rebound’ enhancement of the spinal transmission after exposure to GABA B agonists sheds light on the possible mechanism of the severe withdrawal effects after abrupt termination of baclofen treatment in patients suffering from multiple sclerosis.

In order to specify the role of the periaqueductal grey (PAG) in vocal production, we tested the effect of PAG inactivation on the electrical elicitability of vocalization from various brain structures in the squirrel monkey. For this purpose, we implanted stimulation electrodes at 64 vocalization-eliciting sites throughout the brain and compared the elicitability of vocalization before and after muscimol injection into the PAG. It turned out that only vocalization sites in the forebrain (cingulate cortex, hypothalamus) and rostralmost mesencephalic reticular formation could be blocked by PAG inactivation, whereas all vocalization sites in the caudal midbrain, pons and medulla remained unaffected. It is concluded that the PAG is not the site of vocal pattern generation, but rather serves gating functions.

Ionotropic gamma-aminobutyric acid (GABA) receptors are known to mediate excitation in neonatal neurones as a crucial developmental factor. In the present study we employed calcium imaging techniques with the calcium indicator Fura-2-AM to study the pharmacology of GABA-induced calcium responses in cultures prepared from neonatal rat superficial superior colliculus (SC), after immunocytochemical labelling confirmed the presence of GABA(C) rho(1) subunits in 35% of neurones. Rises in neuronal intracellular calcium were obtained in response to GABA and also to the subtype-specific GABA(A) agonist 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol. However, the GABA(C) agonist cis-4-aminocrotonic acid induced calcium response only at unspecifically high concentrations (500 microM). Co-application of GABA antagonists revealed that both GABA(AC) agonists' actions could be blocked by the GABA(A) antagonist bicuculline but not the GABA(C) antagonists 1,2,5,6-tetrahydro-(pyridin-4-yl) methylphosphinic acid. This suggests that activation of GABA(A) but not GABA(C) receptors contributes to excitatory GABA responses and related calcium signals in neonatal SC neurones.

The effects of bath applied muscimol upon spontaneous and evoked antidromic activity recorded from lumbar dorsal roots was investigated in hemisected, isolated preparations of rat spinal cord. In magnesium free medium containing 0.1 microM 4-aminopyridine, bursts of high amplitude (up to 1 mV), dorsal root reflexes were recorded. These were blocked by low concentrations of muscimol (2-5 microM). Higher concentrations (5-20 microM) of muscimol caused a concentration-dependent increase in the frequency of small amplitude (

Regulation of striatal dopamine release by gamma-aminobutyric acid (GABA) neurotransmission was investigated using voltammetry in freely moving rats, following focal injection of the GABA B receptor agonist baclofen or the GABA B receptor antagonist 5-aminovaleric acid (5-AVA) in either the substantia nigra pars reticulata (SNr) or the substantia nigra pars compacta (SNc). Administration in the SNr of baclofen and 5-AVA at the dose of 2 pg, but not of 0.2 pg, resulted in a decrease and an increase in striatal dopamine release, respectively. In contrast, when injected in the SNc, 5-AVA only produced a transient increase in striatal dopamine release, while baclofen remained ineffective. This suggests that GABA B receptors in the SNr, but not the SNc, may play a major role in the control of nigrostriatal dopamine (DA) activity and the release of DA in the striatum.

It has been reported that GABA(B) receptors are abundantly expressed in the superficial superior colliculus (sSC). To elucidate the action of GABA(B) receptors in the sSC, we tested the effect of a specific GABA(B) receptor agonist baclofen on sSC neurons in slices obtained from rats (16-22 days postnatal) using the whole-cell patch clamp technique. Bath application of baclofen (10-30 microM) elicited hyperpolarization associated with an increase in potassium conductance in morphologically different types of neurons including putative excitatory and inhibitory neurons. In addition, baclofen presynaptically suppressed electrically induced excitatory postsynaptic currents and inhibitory postsynaptic currents. These effects of baclofen were antagonized by a specific GABA(B) receptor antagonist CGP55845A (1 microM). The results suggest that the GABA(B) receptors have an inhibitory action at both post- and pre-synaptic sites in the sSC.

Immobilization stress increased the number of neurons with Fos immunoreactivity, mainly in the ventral zone of the rostral part of the lateral septal nucleus (LSV) in rats. Immobilization stress caused an increase in blood pressure, and the stress-induced pressor response was inhibited by the GABA(A) receptor agonist, muscimol (8 and 80 pmol), injected bilaterally into the rostral part of the LSV in a dose-dependent manner. Intracerebroventricular injection of muscimol (16 pmol) did not affect the immobilization stress-induced pressor response. These findings suggest that the rostral part of the LSV is involved in mediation of the stress-induced pressor response.

The modulation of GABAergic transmission by metabotropic glutamate (mGlu) receptors was examined in histaminergic neurons using a conventional whole-cell patch clamp technique. DHPG, a selective group I mGlu receptor agonist, had no effect on GABAergic inhibitory postsynaptic currents (IPSCs). However, DCG-IV (1 μМ) and L-AP4 (1 μМ), selective group II or III mGlu receptor agonists, respectively, decreased the amplitude of GABAergic IPSCs and simultaneously increased the paired-pulse ratio. The inhibitory effect of DCG-IV was completely blocked by 200 nM LY341495, a group II and III mGlu receptor antagonist, and the inhibitory effect of L-AP4 was completely blocked by 10 μМ LY341495. These results suggest that multiple mGlu receptors are involved in regulating the excitability of histaminergic neurons.

Male rats show 4-6 penile erection episodes when put in the presence of an inaccessible receptive female. These non-contact penile erections were reduced dose-dependently by muscimol, a gamma aminobutyric acid (GABA)(A) receptor agonist, when given into the paraventricular nucleus of the hypothalamus (0.1, 0.5, 1 and 2 microg). In contrast, baclofen, a GABA(B) receptor agonist (2 microg) was ineffective. Muscimol reduction of non-contact penile erections was not seen when male rats were pretreated with bicuculline methiodide (2 microg) given 5 min before muscimol into the paraventricular nucleus. Since muscimol injected into the paraventricular nucleus also prevents penile erection induced by drugs (e.g. apomorphine, oxytocin or N-methyl-D-aspartic acid), the present results show that an increased GABAergic activity in the paraventricular nucleus can impair the expression of penile erection induced not only by drugs but also by sexual physiological stimuli.

Recent experimental and clinical studies clearly suggest the role of gamma-aminobutyric acid (GABA) in the pathogenesis of tremors. The present study was undertaken to investigate the effect of baclofen, a GABA B receptor agonist on harmaline induced tremors. Four groups of female Wistar rats weighing 100±15 g were injected with harmaline (10 mg/kg, intraperitoneally) for inducing experimental tremors. The animals in groups 2, 3 and 4 were given baclofen by gavage at doses of 2.5, 5 and 10 mg/kg, respectively, half an hour before harmaline administration, whereas, the rats in group 1 served as control and received water. The latency of onset, intensity and duration of tremor and electromyographic (EMG) responses were recorded. Treatment with baclofen resulted in a dose dependent decrease in the intensity of tremor. Our EMG study also revealed a significant decrease in the amplitude of tremors in baclofen treated rats. A highly significant increase in latency of onset of tremor was observed in the rats treated with high dose (10 mg/kg) of baclofen only. This study clearly suggests beneficial effects of baclofen in harmaline induced tremors.

Modulation of excitatory synaptic transmission by agonists for several neurotransmitter receptors was investigated at intrinsic cortical synapses derived from single presynaptic neurons. Excitatory postsynaptic currents (EPSCs) were recorded from layer 5 pyramidal neurons in the rat visual cortex in response to minimal stimulation within the same layer. 5-hydroxytryptamine, adenosine, baclofen, carbachol and DCG-IV all suppressed EPSCs with an increase in paired-pulse ratio. These agonists reduced the frequency of miniature EPSCs without significantly affecting their amplitude distribution. These results suggest that glutamatergic excitatory transmission in the neocortex is under the control of presynaptic inhibition mediated by multiple neuromodulator receptors co-expressed in single presynaptic terminals.

A multichannel glutamate sensor was fabricated that consists of enzyme modified electrodes and has a high sensitivity and selectivity to glutamate. We placed a rat hippocampal slice on the sensor and monitored the current at four electrodes resulting from the stimulation with muscimol, a gamma-aminobutyric acid(A) (GABA(A)) receptor agonist. We obtained different glutamate concentration increases at the different positions, suppressed by bicuculline, a GABA(A) receptor antagonist. This demonstrated that the sensor can monitor the glutamate released via GABA(A) receptors pathways, and the difference in the concentrations may indicate differences in the distribution of GABA(A) receptor as well as diverse receptor functions. This multichannel sensor may be useful for non-invasive, real-time monitoring of glutamate distribution, which would make it a valuable tool for pharmacological analysis.

The weaver mutation causes cell loss in the center of the substantia nigra, pars compacta. We compared the depression of γ-aminobutyric acid (GABA) A synaptic currents by the GABA B agonist R-baclofen in pars compacta neurons of weaver mice which were largely spared from cell degeneration and of wild-type mice. In weaver neurons the suppression of GABA A synaptic currents by R-baclofen was reduced compared to wild-type neurons. The EC 50 of R-baclofen was 6.3 times higher in weaver than in wild-type mice. In the neostriatum, which is not a target of the mutation, such a difference did not exist. We conclude that in the pars compacta the weaver mutation leads to a reduced presynaptic autoinhibition through GABA B receptors which may promote survival of a subset of weaver neurons in the pars compacta.

Nociceptin (NC) and the opioid receptor like-1 receptors are widely distributed in areas of the neuraxis that are part of the descending modulatory pain system. We used the tail-flick assay in mice to assess the interaction between NC and other analgesic compounds acting on different areas of the descending pathway. Given by intracerebroventricular injection, NC induced hyperalgesia at 10 nmol (39% of reduction vs. control group). The same dose of NC reversed analgesia induced by distinct classes of analgesia-producing compounds such as morphine, dynorphin A or baclofen. NC caused a reduction of their antinociceptive effects: 61, 41 and 27%, respectively. Thus, NC at the supraspinal level appears to interact with both opioid and gamma-aminobutyric acid(B) systems producing anti-analgesic effects probably through the descending pathway for pain control.

Using in vitro autoradiography to measure binding of the gamma-aminobutyric acid(A) (GABA(A)) receptor agonist, muscimol, we examined male and female rats on postnatal days 1, 5, 10, and 20. There were no sex differences in muscimol binding in any hypothalamic or limbic regions examined. However, all regions exhibited a developmental increase in the density of binding, except the ventromedial nucleus (VMN) of the hypothalamus. We have previously shown that the adult VMN is the only hypothalamic nucleus containing an abundance of the alpha2 subunit of the GABA(A) receptor and lack of the alpha1 subunit. We hypothesize that the lack of alpha1 may be causally related to the lack of increase in muscimol binding.

Long-term depression has recently been shown to occur at glutamatergic synapses in the avian hippocampus and requires activation of calcium/calmodulin-dependent protein kinase II in the nerve terminal. Here using whole cell and intracellular recordings from brain slices, we show that the N-type calcium channel contributes significantly to glutamate release in the avian hippocampus. Activation of the metabotrobic gamma-aminobutyric acid (GABA)(B) receptor by the specific agonist baclofen blocks synaptic transmission. The action of baclofen was associated with a change in paired pulse facilitation indicating that it resulted from a reduction in the probability of transmitter release. In contrast, no change in paired pulse facilitation was observed following the induction of long-term depression. These results show that activation of GABA(B) receptors and long-term depression reduce transmitter release by distinct mechanisms.

Population spikes associated with the paired pulse facilitation paradigm have been successfully used to measure presynaptic inhibition in several systems. In the present work, this paradigm was used to evaluate the action of baclofen on neostriatal glutamatergic transmission. Baclofen enhanced synaptic facilitation with an EC(50)=0.57 microM and a maximal effect of 457%. Selective antagonists for N-, P- and Q-type Ca(2+)-channels enhanced paired pulse facilitation; suggesting that these channel types participate in the release of transmitter. Nevertheless, neither 1 microM omega-conotoxin GVIA, nor 20 nM omega-agatoxinTK occluded the action of baclofen. Baclofen's action was occluded only by 400 nM omega-agatoxinTK. These data suggest that Q-type Ca(2+)-channels mediate gamma-aminobutyric acid(B) presynaptic inhibition of neostriatal afferents.

The effect of muscimol and baclofen injected into the paraventricular nucleus of the hypothalamus on penile erection and yawning induced by apomorphine, oxytocin and N-methyl-D-aspartic acid (NMDA) was studied in male rats. Muscimol (20-200 ng), but not baclofen (200 ng), injected into the paraventricular nucleus of the hypothalamus 10 min before apomorphine (50 ng), oxytocin (10 ng) or NMDA (50 ng) reduced penile erection and yawning induced by the above compounds given into the paraventricular nucleus. Bicuculline (250 ng) injected into the paraventricular nucleus 5 min before muscimol (100 ng) prevented the inhibitory effect of muscimol on penile erection and yawning induced by apomorphine, oxytocin and NMDA. The present results show that gamma-aminobutyric acid (GABA) inhibits penile erection and yawning by acting on GABA(A) receptors in the paraventricular nucleus of the hypothalamus.

Inhibition of the subthalamic nucleus (STN) has been shown to suppress seizures in different animal models of epilepsy. The aim of this study was to examine the role of the pallidal inputs to the STN in the control of absence seizures in a genetic model in the rat. Disinhibition of the globus pallidus or the ventral pallidum, by local injections of a GABA A antagonist, suppressed absence seizures. Conversely, inhibition of the ventral pallidum by a GABA A agonist aggravated absence seizures. Furthermore, the antiepileptic effects of intrapallidal injections of a GABA A antagonist were correlated with a decrease of extracellular levels of glutamate in the substantia nigra. Our results show that both the globus pallidus and the ventral pallidum exert a modulatory influence on absence seizures and suggest that these effects are mediated through the STN.

The purpose of this study was to determine the involvement of supraspinal centers and spinal synaptic integration in cutaneous vasodilation mediated by dorsal spinal cord stimulation (DCS). Laser Doppler flowmetry was used to assess cutaneous blood flow changes in the rat hindpaw during DCS with a unipolar ball electrode placed at the L2-L3 spinal level. Results demonstrated that transecting the spinal cord at the T10 spinal segment did not alter the DCS response while T13 spinal transection abolished the DCS-induced vasodilation. Inhibition of synaptic activity with topical application of muscimol (0.2 mM) on the dorsal surface of the spinal cord markedly attenuated the DCS response. In conclusion DCS-induced vasodilation involved synaptic integration but did not require input from rostral spinal sites or supraspinal areas.

The possible voltage-dependence of the dopamine transporter (DAT) was investigated using electrophysiological and electrochemical recordings in rat brain slices containing the substantia nigra (SN). Whole-cell patch clamp recordings of DA neurons, revealed that addition of 15 mM KCl rapidly depolarized the membrane potential by approximately 20 mV, whereas these cells were hyperpolarized approximately 10 mV by DA (10 microM) and approximately 14 mV by the GABAB-receptor agonist baclofen (30 microM). High-speed chronoamperometric recordings were used to monitor clearance properties of exogenously applied DA signals during similar manipulations. Superfusion of slices with 15 mM KCl significantly increased the time course of the DA signal, consistent with inhibition of DAT activity. Application of the D2 DA-receptor antagonist sulpiride (50 microM) also significantly increased the time course, suggesting that DA-induced hyperpolarization enhances DAT activity. Baclofen reversed the effects of sulpiride on DA clearance. These results demonstrate that changes in DA cell membrane potential can modulate DAT activity.

Phrenic motoneurons (PMNs) receive intermittently glutaminergic inspiratory drives and GABAergic inhibition in adult mammals. Since gamma-amino-butyric acid (GABA) might act as an excitatory amino acid in early stages of development, we here investigated if GABA(A) receptors inhibit PMNs in neonates. Using in vitro neonatal rat brainstem-spinal cord preparation, local application of GABA and muscimol (a GABA(A) receptor agonist) to the vicinity of PMNs consistently reduced the inspiratory activity of C4 ventral roots. Under whole-cell patch-clamp conditions and in the presence of 0.5 microM TTX to block synaptic transmission, muscimol (10 microM) decreased whole-cell input resistance, and surprisingly, when PMNs were voltage-clamped at their resting membrane potential, muscimol induced depolarizing-inward, rather than hyperpolarizing-outward membrane current. Our findings indicate that GABA(A) receptors mediate a depolarizing blockade of the glutaminergic excitatory inputs to neonatal rat PMNs.

In rats with abnormally low withdrawal thresholds ('allodynia') in one hindpaw induced by a photochemical sciatic lesion, an intrathecal catheter was inserted to the lumber enlargement and an epidural electrode was implanted at T11. I.t. administration of GABA(B) or adenosine A1 receptor agonists (baclofen, R(-)-N6-(2-phenylisopropyl)adenosine (R-PIA)) suppressed allodynia in a dose-dependent fashion. When the two agonists were given together, each in an ineffective dose, there was a normalization of the thresholds. Rats, in which spinal cord stimulation (SCS) could not suppress the allodynia (non-responders), were transformed into SCS-responders by injection of baclofen and R-PIA in low, ineffective doses, combined with SCS. In SCS responding rats, combination of a selective GABA(B) and an adenosine A1 receptor antagonist (CGP 55845, CPT) in low, ineffective doses abolished the SCS-induced threshold normalization. These results indicate that GABAergic and adenosine-dependent mechanisms are involved in the SCS effect and further suggest a strategy for enhancing the therapeutic efficacy of SCS.

The benzodiazepine agonist, diazepam, inhibits cAMP production in the rat brain. Since cAMP influences c-fos activity, we examined the effects of diazepam on expression of this immediate early gene, as indicated by Western blot analysis. Intraperitoneal administration of diazepam increased Fos protein levels in the striatum, but not in the hippocampus. In contrast, pretreatment with diazepam blocked the potent inducing effect of amphetamine, in both brain regions. Similar induction and blockade effects were also observed for a 90 kDa Fos related antigen (Fra), in the striatum and hippocampus. The possible mechanisms underlying the modulatory effects of diazepam on c-fos expression in the brain are discussed.

Studies have demonstrated that median preoptic nucleus (MnPO) neurons play a role in organizing the cardiovascular responses induced by changes in the circulating blood volume. The present study examined whether the MnPO controls cardiovascular function. Male Wistar normotensive (NT) rats and spontaneously hypertensive rats (SHRs; 250-300 g) were anesthetized with urethane (1.2 g kg(-1), i.v.) and instrumented for recordings of mean arterial blood pressure (MAP) and renal blood flow (RBF). The renal vascular conductance (RVC) was calculated as the RBF:MAP ratio and was expressed as a percentage of the baseline value. In the NT rats (n=6), MnPO inhibition produced a MAP reduction (-8.1±1.1 mmHg, p0.05). In the SHRs (n=6), the MAP response to MnPO inhibition was significantly greater (-22.3±4 mmHg, p0.05) than in the NT rats. Furthermore, the increase in the RVC was higher in the SHRs (10.9±3.3%, p0.05). Histological analyses confirmed that the injection sites were confined to the MnPO. We conclude that the MnPO is involved in the tonic regulation of blood pressure in NT rats. Moreover, the greater cardiovascular response to MnPO inhibition observed in the SHRs strongly suggests that the MnPO may contribute to the pathophysiology of essential hypertension.

The effect of gamma-hydroxybutyric acid (GHB) on the excitatory postsynaptic potential (EPSP) evoked in thalamocortical neurones of the rat dorsal lateral geniculate nucleus and ventrobasal thalamus was investigated in vitro. GHB (0.1-5 mM) dose-dependently and reversibly decreased (36-78%) the amplitude of the sensory EPSP. This effect of GHB was blocked by the GABAB receptor antagonist CGP 35348 (1 mM). NCS 382 (1-3 mM), a putative GHB receptor antagonist, did not antagonise but weakly potentiated both the GHB- and baclofen-mediated decrease of the EPSP amplitude.

In vitro brainstem-spinal cord preparations isolated from neonatal (0-5 days old) rats were used to investigate the GABAB receptor-mediated modulation of the dorsal root evoked potentials in lumbar motoneurons recorded intracellularly. The GABAB receptor agonist, baclofen, at low concentrations (1-10 microM), caused a reduction of the amplitude of the monosynaptic excitatory postsynaptic potential (EPSP), in a concentration-dependent manner. The depression of EPSPs was likely exerted at a presynaptic level since it occurred without any significant change of the passive membrane properties of the motoneurons. The two GABAB receptor antagonists, CGP 35348 and CGP 55845A blocked the effects of baclofen. These two compounds may be useful tools to study the evolution of GABAB receptor-mediated presynaptic inhibition during ontogenesis.