GABAB Agonist Research Articles

Expression of gamma-aminobutyric acid receptor (subtype A) in prostate cancer

Background. In prostate cancer, gamma-aminobutyric acid (GABA) has been previously reported to increase cellular proliferation via the ionotropic GABAa receptor (GABAar) and to promote cellular invasiveness via the metabotropic GABAb receptor. Methods. In this study, we have investigated, by immunohistochemistry, GABAar levels in 12 normal human prostate, 13 benign prostatic hyperplasia (BPH) and 148 human prostate cancer specimens. We have also examined the effect of several GABA agonists and antagonists on the in vitro proliferation of four human prostate cancer cell lines: LNCaP, MDA-PCA-2b, DU145 and PC3. Results. GABAar immunoreactivity was present in the stroma of ∼75% of the normal and BPH specimens, and in 95% of the prostate cancer specimens. Also, low to moderate GABAar staining was observed in the acinar epithelium of 50 (33%) prostate cancer specimens. No correlation was observed between GABAar staining and patient age, Gleason Sum or TNM stage. A GABAa agonist isoguvacine, at doses between 5–50 ug/ml (31–310 uM), stimulated the proliferation of all four human prostate cancer cell lines, tested. Baclofen, a GABAb agonist (up to 50 ug/ml, 234 uM) had no effect on growth. Also, at concentrations up to 100 ug/ml, GABA antagonists, bicuculline (223 uM), picrotoxin (166 uM) and saclofen (400 uM), did not have significant growth-inhibitory effects. However, dihydroergotoxine, which binds the GABAar chloride ion-channel, inhibited cellular proliferation (IC50 18–38 uM). Conclusions. These data indicate frequent expression of GABAar in prostate cancer and support a role for GABAar in the proliferation of prostate cancer cells.

Neuroprotection Afforded by GABA Against Oxygen-Glucose Deprivation-Induced Injury in Rat Cortical Brain Slices: An Hormetic Dose-Response Effect

The aim of the present investigation was to assess neuroprotection exerted by GABA (100 nM-100 mM) in rat cortical brain slices subjected to oxygen-glucose deprivation and reoxygenation. Neuronal injury and neuroprotection were assessed by measuring the release of glutamate and lactate dehydrogenase and tissue water content. Results demonstrate that GABA exerted neuroprotective effects according to a U-shaped, hormetic-like, concentration-response curve, with an efficacy window of 10-100 I¼M concentration. In order to verify whether GABA-neuroprotective effects were GABAA- or GABAB-mediated, Muscimol (GABAA agonist) and Baclofen (GABAB agonist) were tested. Results demonstrated that only Muscimol exerted neuroprotection according to a U-shaped, hormetic-like, concentration-response curve, while baclofen was ineffective. In conclusion, the hormetic response of GABA or GABAA agonists should be taken into consideration when designing experiments aimed at assessing neuroprotection by these agents against ischemia/reoxygenation injury.

Growth hormone response to the GABA-B agonist baclofen in 3-week abstinent alcoholics

Gamma-aminobutyric acid (GABA) dysfunction is a known feature of alcoholism. We investigated GABA-B receptor activity in 3-week abstinent alcoholics using the growth hormone (GH) response to baclofen, a GABA-B receptor agonist. The study aimed to investigate the relationship between GABA-B receptor activity and alcohol withdrawal. GH response to baclofen was measured in alcohol-dependent males without depression ( n = 22) who were on day 21 of alcohol abstinence and in healthy control male subjects ( n = 23). After 20 mg baclofen was given orally to the subjects, blood samples for GH assay were obtained every 30 min for the subsequent 150 min. The patients were divided into two subgroups (continuing withdrawal and recovered withdrawal subgroups) according to their withdrawal symptom severity scores on day 21 of alcohol cessation. Baclofen administration significantly altered GH secretion in the controls, but not in the patients. When GH response to baclofen was assessed as ΔGH, it was lower in the patients with continuing withdrawal symptoms than in the controls and in the recovered withdrawal group. Impaired GH response to baclofen in all patients mainly pertained to the patients whose withdrawal symptoms partly continued. Our results suggest that reduced GABA-B receptor activity might be associated with longer-term alcohol withdrawal symptoms in alcoholic patients.

Contribution of Ventral Tegmental GABA Receptors to Cocaine Self-administration in Rats

Recent evidence has suggested that compounds affecting GABAergic transmission may provide useful pharmacological tools for the treatment of cocaine addiction. Using a rat model of self-administration, the present study examined the effects of GABA agonists and antagonists injected directly into the ventral tegmental area (VTA) on cocaine intake in rats trained to self-administer cocaine (0, 125, 250 and 500 microg/infusion) under an FR5 schedule of reinforcement. Separate groups of rats received bilateral intra-VTA injections of the GABA-A antagonist picrotoxin (34 ng/side, n = 7; 68 ng/side, n = 8), GABA-A agonist muscimol (14 ng/side, n = 8), GABA-B agonist baclofen (56 ng/side, n = 7; 100 ng/side, n = 6), picrotoxin (68 ng/side) co-injected with the GABA-B antagonist 2-hydroxysaclofen (100 ng/side, n = 7; 2 microg/side, n = 8) or artificial cerebrospinal fluid (aCSF, n = 6) to assess the effects of the various compounds on the cocaine self-administration dose-response curve. Both picrotoxin and baclofen reduced responding maintained by cocaine, whereas muscimol had no effect on responding. In contrast, neither picrotoxin (n = 6) nor baclofen (n = 8) affected responding maintained by food. Interestingly, 2-hydroxysaclofen effectively blocked the suppression of responding produced by picrotoxin, suggesting that both picrotoxin and baclofen exert their effects via activation of GABA-B receptors. Additionally, these effects appear to be specific to cocaine reinforcement, supporting current investigation of baclofen as a treatment for cocaine addiction.

GABA receptor agonism in the sublenticular central extended amygdala impairs medial forebrain bundle self-stimulation but GABA blockade does not enhance it

The sublenticular central extended amygdala (SLEAc), which is important in medial forebrain bundle (MFB) self-stimulation, is heavily populated with GABAergic medium spiny neurons that intercommunicate via local axon collaterals. This study examines the role of GABAergic communication in the SLEAc in MFB self-stimulation. Male Long–Evans rats were given unilateral MFB stimulation electrodes and bilateral injection guide cannulae aimed at the SLEAc. Changes in the stimulation's reward efficacy and in the animals’ ability to respond for stimulation following 0.5 μl injections of saline, 50 ng of muscimol (GABA-A agonist), 75 ng of baclofen (GABA-B agonist), 25 ng of bicuculline methiodide (GABA-A antagonist), and 500 ng of phaclofen (GABA-B antagonist) were assessed using the rate–frequency curve shift paradigm. Injecting the receptor agonists ipsi- and contralateral to the stimulation site substantially decreased stimulation reward efficacy and responding. Baclofen's effects were more anatomically specific to the SLEAc and baclofen was less challenging to response rate than was muscimol. Ipsi- and contralateral injections of the receptor blockers and saline had no effect on either stimulation reward efficacy or response rate, although the blockers, especially phaclofen, sometimes enhanced response rate when injected into the globus pallidus or ventral pallidum. Comparing these results to others’ observations about GABAergic communication in the basal forebrain suggests that muscimol may have inhibited endemic SLEAc neurons while baclofen may have impaired the release of glutamate into the SLEAc from cortical and amygdalar afferents. These results also suggest functional differences between the two basal forebrain macrostructures of striatopallidum and extended amygdala.

CGP7930: A Positive Allosteric Modulator of the GABAB Receptor

CGP7930 (3-(3',5'-Di-tert-butyl-4'-hydroxy)phenyl-2,2-dimethylpropanol) is a positive allosteric modulator of the metabotropic GABAB receptor. CGP7930 has been found to modulate the GABAB receptor in the open, or high affinity, state increasing agonist affinity for the receptor and signal transduction efficacy following agonist stimulation. The GABAB heteromeric subunit B2, involved in signal transduction but not ligand binding, seems to be the site of action of CGP7930 and similar allosteric modulators. When administered alone in naïve animals, CGP7930 acts as an anxiolytic in rodents without other overt behavioral effects and has also been demonstrated to reduce self-administration of nicotine, cocaine, or alcohol in rodents, suggesting that "fine tuning" of the GABAB receptor by positive allosteric modulators may be able to regulate abuse of these drugs. Baclofen, the GABAB agonist, is currently finding use in treating addiction and various other disorders, but this can result in off-target effects and tolerance. CGP7930 when co-administered with baclofen enhances its potency, which could in theory minimize deleterious effects. Further study of CGP7930 is required, but this compound, and others like it, holds potential in a clinical setting.

Lateral Diffusion of the GABAB Receptor Is Regulated by the GABAB2 C Terminus

GABAB (gamma-aminobutyric acid, type B) is a heterodimeric G-protein-coupled receptor. The GABAB1 subunit, which contains an endoplasmic reticulum retention sequence, is only transported to the cell surface when it is associated with the GABAB2 subunit. Fluorescence recovery after photobleaching studies in transfected COS-7 cells and hippocampal neurons revealed that GABAB2 diffuses slowly within the plasma membrane whether expressed alone or with the GABAB1 subunit. Treatment of cells with brefeldin A revealed that GABAB2 moves freely within the endoplasmic reticulum, suggesting that slow movement of GABAB2 is a result of its plasma membrane insertion. Disruption of the cytoskeleton did not affect the mobility of GABAB2, indicating that its restricted diffusion is not due to direct interactions with actin or tubulin. To determine whether the C terminus of GABAB2 regulates its diffusion, this region of the subunit was attached to the lymphocyte membrane protein, CD2, which then exhibited a slower rate of lateral diffusion. Furthermore, co-expression of a cytoplasmically expressed soluble form of the GABAB2 C terminus increased movement of the GABAB2 subunit. We constructed forms of GABAB2 with various C-terminal truncations. Truncation of GABAB2 after residue 862, but not residue 886, caused a dramatic increase in its mobility, suggesting that the region between these two residues is critical for restricting GABAB2 diffusion. Finally, we investigated whether activation of GABAB might modulate its movement. Treatment of COS-7 cells with the GABAB receptor agonist baclofen significantly increased its mobile fraction. These data show that the restricted movement of GABAB at the cell surface is regulated by a region within its C terminus.

Baclofen for binge eating: An open‐label trial

Baclofen is a GABA-B agonist that may be useful in the treatment of substance use disorders, and also reduces 'binge-like' eating in rodents. We hypothesized that baclofen might be effective in reducing binge eating episodes in binge eating disorder (BED) and bulimia nervosa (BN). Seven women with BED (n = 4) or BN (n = 3) took baclofen (60 mg/day) for 10 weeks. Six out of seven patients completed the full 10-week trial. Five out of seven participants (3 BED; 2 BN) demonstrated 50% or greater reduction of frequency of binge eating from beginning to end of the study. Three out of seven participants (2 BED; 1 BN) were free of binge eating at study end. Four out of seven participants elected to continue baclofen at study end. Baclofen was well tolerated by the participants. In this open-label trial, baclofen was associated with decreased binge eating frequency in patients with BED and BN.

Effects of baclofen on temperature perception in humans

The aim of this study was to evaluate the effect of chronic consumption of the GABAB agonist baclofen on temperature perception in humans. We investigated temperature perception thresholds to detect warm and cold stimuli in a group of 21 patients with spinal cord injury, who were chronically consuming oral baclofen at different daily doses to treat spasticity. Temperature perception thresholds were assessed above the level of the lesion, using a psychophysical approach based on the ability of the subjects to perceive precisely quantified sensory stimuli (quantitative sensory testing, QST). The data were compared with a control group of healthy subjects, not receiving baclofen. We found that chronic baclofen consumption increased temperature perception thresholds for both cold and warm stimuli in a dose-dependent manner. Temperature perception thresholds did not depend on the level of the lesion nor on the duration of baclofen treatment, suggesting that our finding represent normal GABAB-mediated modulation in spared nervous structures. We conclude that GABAB therefore plays a role in temperature perception in humans.

Ameliorating effect of emodin, a constitute of Polygonatum multiflorum, on cycloheximide-induced impairment of memory consolidation in rats

The aim of the present study was intended to investigate the ameliorating effects of emodin on memory consolidation via cholinergic, serotonergic and GABAergic neuronal systems in rats. First, we evaluated the ameliorating effects of emodin on cycloheximide (CXM)-induced impairment of passive avoidance response in rats. Secondly, we clarified the role of cholinergic, serotonergic or GABAergic system on the ameliorating effect of emodin by using 5-HT1A receptor partial agonist, 5-HT2 receptor antagonist, GABAB agonist, GABAA antagonist and muscarinic receptor antagonist. Emodin protected the rat from CXM-induced memory consolidation impairment. The beneficial effect of emodin on CXM-induced memory consolidation impairment was amplified by 8-OH-DPAT (5-HT1A receptor partial agonist) and ritanserin (5-HT2 receptor antagonist), but reduced by scopolamine. These results suggested that the beneficial effect of emodin on CXM-induced memory consolidation impairment was amplified by serotonergic 5-HT1A-receptor partial agonist and 5-HT2 receptor antagonist but reduced by muscarinic receptor antagonist.

Altered synaptic input and GABAB receptor function in spinal superficial dorsal horn neurons in rats with diabetic neuropathy

Hyperactivity of spinal dorsal horn neurons plays an important role in the development of diabetic neuropathic pain. However, little is known as to whether synaptic input to spinal dorsal horn neurons is altered in diabetic neuropathy. Also, the function of GABAB receptors in the control of synaptic input to dorsal horn neurons in diabetes remains poorly understood. To determine the changes in synaptic input to dorsal horn neurons and the GABAB)receptor function in streptozotocin-induced diabetes, we performed whole-cell recording (GDP-beta-S included in the internal solution) on lamina II neurons in rat spinal cord slices. The frequency of glutamatergic mEPSCs and the amplitude of monosynaptic EPSCs evoked from the dorsal root were significantly higher in diabetic than in control rats. On the other hand, the basal frequency and amplitude of GABAergic spontaneous IPSCs and mIPSCs and those of glycinergic spontaneous IPSCs and mIPSCs did not differ significantly between control and diabetic rats. The GABAB agonist baclofen produced a significantly greater reduction in dorsal root-evoked EPSCs and the frequency of mEPSCs in control than in diabetic rats. However, the inhibitory effect of baclofen on GABAergic and glycinergic spontaneous IPSCs and mIPSCs was not significantly different in the two groups. These findings suggest that increased glutamatergic input from primary afferents to dorsal horn neurons may contribute to synaptic plasticity and central sensitization in diabetic neuropathic pain. Furthermore, the function of presynaptic GABAB receptors at primary afferent terminals, but not that on GABAergic and glycinergic interneurons, in the spinal cord is reduced in diabetic neuropathy.

Baclofen alters flash-evoked potentials in Long–Evans rats

This experiment examined the effects of the GABA-B agonist baclofen on flash-evoked potentials (FEPs) recorded from both the visual cortex (VC) and superior colliculus (SC) of chronically implanted male Long–Evans rats. FEPs were recorded at 5, 25, 45, and 65 min following intraperitoneal injections of saline, and of 1.25, 2.5, 5.0, and 10.0 mg/kg baclofen on separate days. In the VC, the amplitude of components P 23, P 37, N 55, N 150, and P 242 increased, while the amplitude of components N 31 and P 48 decreased following baclofen administration. P 88 was unchanged. In the SC, components P 28, N 49, N 55, and N 59 were reduced in amplitude, while P 39 was unaffected by baclofen. These effects on amplitudes were dose- and time-dependent. Many peak latencies in the VC and SC were altered by baclofen, although there was no obvious pattern of change, with some decreasing, a few increasing, and others unchanged. Body temperature was recorded in a separate group of animals, with both the 5.0 and 10.0 mg/kg doses of baclofen producing significant hypothermia. The 10.0 mg/kg dose of baclofen resulted in a significant decrease in movement during the recording sessions, but not in subsequent open field observations. The results show the involvement of GABA-B receptors in the production/modulation of the various components of FEPs.

In vivo temporal property of GABAergic neural transmission in collateral feed-forward inhibition system of hippocampal–prefrontal pathway

Anatomical evidence suggests that rat CA1 hippocampal afferents collaterally innervate excitatory projecting pyramidal neurons and inhibitory interneurons, creating a disynaptic, feed-forward inhibition microcircuit in the medial prefrontal cortex (mPFC). We investigated the temporal relationship between the frequency of paired synaptic transmission and gamma-aminobutyric acid (GABA)ergic receptor-mediated modulation of the microcircuit in vivo under urethane anesthesia. Local perfusions of a GABAa antagonist (−)-bicuculline into the mPFC via microdialysis resulted in a statistically significant disinhibitory effect on intrinsic GABA action, increasing the first and second mPFC responses following hippocampal paired stimulation at interstimulus intervals of 100–200 ms, but not those at 25–50 ms. This (−)-bicuculline-induced disinhibition was compensated by the GABAa agonist muscimol, which itself did not attenuate the intrinsic oscillation of the local field potentials. The perfusion of a sub-minimal concentration of GABAb agonist ( R)-baclofen slightly enhanced the synaptic transmission, regardless of the interstimulus interval. In addition to the tonic control by spontaneous fast-spiking GABAergic neurons, it is clear the sequential transmission of the hippocampal-mPFC pathway can phasically drive the collateral feed-forward inhibition system through activation of a GABAa receptor, bringing an active signal filter to the various types of impulse trains that enter the mPFC from the hippocampus in vivo.

Dissociation and trafficking of rat GABAB receptor heterodimer upon chronic capsaicin stimulation

Gamma-aminobutyric acid type B receptors (GABAB) are G-protein-coupled receptors that mediate GABAergic inhibition in the brain. Their functional expression is dependent upon the formation of heterodimers between GABAB1 and GABAB2 subunits, a process that occurs within the endoplasmic reticulum. However, the mechanisms that regulate GABAB receptor oligomerization at the plasma membrane remain largely unknown. We first characterized the functional cytoarchitecture of an organotypic co-culture model of rat dorsal root ganglia and spinal cord. Subsequently, we studied the interactions between GABAB subunits after chronic stimulation of sensory fibres with capsaicin. Surface labelling of recombinant proteins showed a decrease in subunit co-localization and GABAB2 labelling, after capsaicin treatment. In these conditions, fluorescence lifetime imaging measurements further demonstrated a loss of interactions between green fluorescent protein-GABAB1b and t-dimer discosoma sp red fluorescent protein-GABAB2 subunits. Finally, we established that the GABAB receptor undergoes clathrin-dependent internalization and rapid recycling to the plasma membrane following activation with baclofen, a GABAB agonist. However, in cultures chronically stimulated with capsaicin, the agonist-induced endocytosis was decreased, reflecting changes in the dimeric state of the receptor. Taken together, our results indicate that the chronic stimulation of sensory fibres can dissociate the GABAB heterodimer and alters its responsiveness to the endogenous ligand. Chronic stimulation thus modulates receptor oligomerization, providing additional levels of control of signalling.

A GABAB agonist reverses the behavioral sensitization to morphine in rats

In laboratory animals, repeated administration of drugs of abuse causes sensitization to their stimulant and rewarding effects. Neuroadaptations underlying sensitization could be related to those that contribute to addictive behaviors. An increased understanding of the molecular mechanisms of sensitization could lead to improved treatments for addiction. Since baclofen (BCF) co-administration has been reported to block the development and the expression of motor sensitization to morphine (MOR), the present study examined the hypothesis that a chronic treatment with BCF alone might reverse and/or prevent MOR-induced sensitization. Rats were first sensitized to MOR (saline or 10 mg/kg MOR i.p.; days 1-10) and then chronically treated with BCF (saline or 2 mg/kg BCF i.p.; days 11-20). Finally, the motility effect of MOR (10 mg/kg i.p.) was assessed 3 and 30 days after the end of BCF treatment. The same rats were again challenged with MOR on day 70, after a further period of saline or MOR treatment (days 51-60). Behavioral sensitization to MOR was observed in control animals but not in rats chronically treated with BCF (days 23 and 50). Thus, BCF completely reversed MOR-induced sensitization, and its effect was long lasting. However, a previous repeated BCF treatment did not prevent the development of sensitization to MOR both in naive and desensitized rats. The present results confirm that gamma-aminobutyric acid (GABA)(B) receptors play an important role in the expression of motor sensitization to MOR and suggest that GABA(B) agonists could be useful for reversing the neuroadaptations related to drug addiction.

Effects of GABAA and GABAB agonists on interhemispheric inhibition in man

ObjectiveAnimal studies on neurotransmitter systems that mediate interhemispheric inhibition (IHI) suggest that, (i) callosal transmission is regulated by presynaptic GABAB receptors, and (ii) GABAA-ergic neurones mediate early IHI, whereas GABAB-ergic neurones mediate later IHI. In humans the mechanism is unclear. Interactions between cortical inhibitory circuits suggest a postsynaptic GABAB-ergic mechanism. We will here test this hypothesis. MethodsShort-latency IHI (s-IHI) and long-latency IHI (l-IHI) were evaluated using the paired pulse paradigm before and under medication with (i) a GABAB-agonist (baclofen) in 17 subjects, and (ii) a GABAA-agonist (midazolam) in 10 subjects participating twice. ResultsBaclofen did not significantly enhance s-IHI. L-IHI between 20 and 50ms was significantly strengthened, and obtained also at ISIs between 100 and 200ms. Midazolam had no effect on s-IHI, whereas l-IHI was attenuated. ConclusionsOur results support the hypothesis, that l-IHI in humans is mediated by postsynaptic GABAB receptors. GABAA-ergic medication resulted in attenuation of l-IHI. Regarding s-IHI, our results are inconclusive and require further investigation. SignificanceThis is the first human study evaluating the effect of baclofen on IHI, indicating that l-IHI is mediated by GABAB-ergic neurones. Because interhemispheric interaction is now also been used as a therapeutic approach, understanding the underlying neurotransmitter systems will be increasingly relevant.

XP19986 Decreases Reflux and Is Well Tolerated in GERD Patients

Purpose: XP19986 is a prodrug of R-baclofen, a GABA-b agonist. The primary objective of this study is to assess the efficacy and tolerability of a controlled-release formulation of XP19986 for decreasing the number of reflux episodes in patients with GERD. Methods: This is a multi-center, randomized, double-blind, placebo-controlled, crossover, study. Separate cohorts of patients are enrolled at each of four escalating dose levels. Enrolled patients have a history of GERD symptoms at least 3 times per week and ≥ 20 reflux episodes on impedance-pH monitoring over 2 hrs following a high-fat meal, and receive single doses of XP19986 or placebo in test periods separated by 4 to 7 days. High-fat meals are consumed at 2 and 6 hrs after dosing. Reflux episodes recorded and blood samples for XP19986 and R-baclofen levels collected through 12-hours post-dose. Other measures include GERD symptoms, vital signs, ECG, and clinical labs. Results: The primary endpoint for the clinical trial was the total number of reflux events over the 12-hour monitoring period following the dose of XP19986. For the combined 10, 20 and 40 mg groups (N = 35), the median number of reflux events during placebo treatment was 51.0, and there were fewer reflux events during XP19986 (median difference 7.0; Wilcoxon signed-rank p= 0.034). For the individual 10, 20 and 40 mg dosage groups, the median number of reflux events during placebo treatment was 79.0 (N = 12), 52.0 (N = 12) and 34.0 (N = 11), respectively. The median reduction during XP19986 treatment compared to placebo was 2.0 (p= 0.458), 11.5 (p= 0.210) and 10.0 (p= 0.051), respectively. In the 40 mg dose group, 9 of 11 patients had fewer reflux events after XP19986 treatment than after placebo treatment. XP19986 was well tolerated at all dose levels: no adverse event occurred in more than one patient on XP19986. Fatigue occurred in 1 patient each on placebo and XP19986. Nausea occurred in 3 patients on placebo and none on XP19986, headache in 3 on placebo and none on XP19986. Conclusions: The results for three dose levels of XP19986 are encouraging, both in terms of the efficacy in reducing reflux and the favorable tolerability of XP19986 in patients with GERD. Data for the 60 mg dose group and pharmacokinetic results will be available at the time of the meeting.

Target-Specific Regulation of Synaptic Efficacy in the Feeding Central Pattern Generator ofAplysia:Potential Substrates for Behavioral Plasticity?

The contributions to this symposium are unified by their focus on the role of synaptic plasticity in sensorimotor learning. Synaptic plasticities are also known to operate within the central pattern generator (CPG) circuits that produce repetitive motor programs, where their relation to adaptive behavior is less well understood. This study examined divergent synaptic plasticity in the signaling of an influential interneuron, B20, located within the CPG that controls consummatory feeding-related behaviors in Aplysia. Previously, B20 was shown to contain markers for catecholamines and GABA (Díaz-Ríos et al., 2002), and its rapid synaptic signaling to two follower motor neurons, B16 and B8, was found to be mediated by dopamine (Díaz-Ríos and Miller, 2005). In this investigation, two incremental forms of increased synaptic efficacy, facilitation and summation, were both greater in the signaling from B20 to B8 than in the signaling from B20 to B16. Manipulation of the membrane potentials of the two postsynaptic motor neurons did not affect facilitation of excitatory postsynaptic potentials (EPSPs) to either follower cell. Striking levels of summation in B8, however, were eliminated at hyperpolarized membrane potentials and could be attributed to distinctive membrane properties of this postsynaptic cell. GABA and the GABAB agonist baclofen increased facilitation and summation of EPSPs from B20 to B8, but not to B16. The enhanced facilitation was not affected when the membrane potential of B8 was pre-set to hyperpolarized levels, but GABAergic effects on summation were eliminated by this manipulation. These observations demonstrate a target-specific amplification of synaptic efficacy that can contribute to channeling the flow of divergent information from an intrinsic interneuron within the buccal CPG. They further suggest that GABA, acting as a cotransmitter in B20, could induce coordinated and target-specific pre- and postsynaptic modulation of these signals. Finally, we speculate that target-specific plasticity and its modulation could be efficient, specific, and flexible substrates for learning-related modifications of CPG function.

Morphine withdrawal syndrome and its prevention with baclofen: Autoradiographic study of μ‐opioid receptors in prepubertal male and female mice

Although the expression of the morphine (MOR) withdrawal syndrome is more marked in male mice than in females, we have demonstrated that the GABAB agonist baclofen (BAC) is able to attenuate MOR withdrawal signs in either sex. In order to extend these previous observations, the aim of the present study was to evaluate the mu-opioid receptor labeling in various brain areas in mice of either sex, during MOR withdrawal and its prevention with BAC. Prepubertal Swiss-Webster mice were rendered dependent by intraperitonial (i.p.) injection of MOR (2 mg/kg) twice daily for 9 days. On the 10th day, dependent animals received naloxone (NAL; 6 mg/kg, i.p.) 60 min after MOR, and another pool of dependent mice received BAC (2 mg/kg, i.p.) previous to NAL. Thirty minutes after NAL, mice were sacrificed and autoradiography with [3H]-[D-Ala2, N-Me-Phe4, -glycol5] enkephalin (DAMGO) was carried out on mice brains at five different anatomical levels. Autoradiographic mapping showed a significant increase of mu-opioid receptor labeling during MOR withdrawal in nucleus accumbens core (NAcC), caudate putamen (CPu), mediodorsal thalamic nucleus (MDTh), basolateral and basomedial amygdala, and ventral tegmental area vs. respective control groups in male mice. In contrast, opiate receptor labeling was not significantly modified in any of the brain areas studied in withdrawn females. BAC reestablished mu-opioid receptor binding sites during MOR withdrawal only in NAcC of males, and a similar tendency was observed in CPu and MDTh, even when it was not statistically significant. The sexual dimorphism observed in the present study confirms previous reports indicating a greater sensitivity of males in response to MOR pharmacological properties. The present results suggest that the effect of BAC in preventing the expression of MOR withdrawal signs could be related with the ability of BAC to reestablish the mu-opioid receptor labeling in certain brain areas.

Electrically stimulated relaxation is not mediated by GABA in cat lower esophageal sphincter muscle

This study examined the effect of Gamma-Amino butyric acid (GABA) and selective GABA receptor related drugs on the electrically stimulated relaxation in the lower esophageal sphincter muscle (LES) of a cat. Tetrodotoxin (10(-6) M) suppressed the electrically stimulated (0.5-5 Hz) relaxation of the LES. However, guanethidine (10(-6) M) and atropine (10(-6) M) had no effect indicating that the relaxations were neurally mediated via the nonadrenergic and noncholinergic (NANC) pathways. NG-nitro-L-arginine methyl ester (10(-4) M, L-NAME) also inhibited the relaxant response but did not completely abolish the electrically stimulated relaxation with 60 % inhibition, which suggests the involvement of nitric oxide as an inhibitory transmitter. This study examined the role of GABA, an inhibitory neurotransmitter, on neurally mediated LES relaxation. GABA (10(-3)-10(-5) M, non selective receptor agonist), muscimol (10(-3)-10(-5) M, GABA-A agonist), and baclofen (10(-3)-10(-5) M, GABA-B agonist) had no significant effect on the electrically stimulated relaxation. Moreover, bicuculline (10(-5) M, GABA-A antagonist) and phaclofen (10(-5) M, GABA-B antagonist) had no inhibitory effect on the electrically stimulated relaxation. This suggests that GABA and the GABA receptor are not involved in the electrically stimulated NANC relaxation in the cat LES.