Inhibitory Effect Of GABA Research Articles

1470-P: GABA Stimulates or Inhibits Insulin and Somatostatin Secretion in Human Pancreatic Islets Depending on Glucose Concentration

GABA is an inhibitory neurotransmitter synthesized and secreted by β-cells that is reported to modulate glucagon, somatostatin (SST) , and insulin secretion. However, the direction of the effect of GABA, whether stimulatory or inhibitory, on insulin secretion is controversial. Our results from human islets show that GABA is inhibitory to insulin secretion under glucose-stimulating conditions. Further, islets from conditional knockout mice lacking the GABA-synthesizing enzymes GAD65 and GAD67 oversecrete insulin during glucose stimulation, consistent with GABA being inhibitory. However, the inhibitory effect of GABA is somewhat paradoxical as ligation of GABA-A receptor Cl channels are expected to depolarize the β-cell that should contribute to stimulating insulin secretion. It is also unclear if GABA signaling in β-cells is strictly autocrine or if the observed inhibition is mediated through an indirect effect of inducing SST secretion which, in turn, inhibits β-cells. Here, we improve understanding of GABA’s effect on insulin secretion by transiently applying 100 μM of GABA to human islets during perifusion with low (3 mM) and high glucose (16.7 mM) . GABA stimulated both insulin and SST in low glucose but inhibited insulin and SST when applied in high glucose. Thus, the modulation of GABA presents both stimulatory and inhibitory effects on β- and δ-cells depending on glucose concentration. In addition, the effect of GABA on β-cells appears to be direct and independent of SST signaling because SST and insulin secretion both change in the same direction with applied GABA. These results suggest the hypothesis that β-cell GABA-A receptors convey stimulatory responses to GABA under low glucose, but that G-protein coupled GABA-B receptors mediate inhibitory GABA signaling under high glucose. This mechanism would provide a consensus model that reconciles decades of seemingly antithetical studies where GABA is either inhibitory or stimulatory. Disclosure S.Ferreira: None. D.W.Hagan: None. E.Phelps: Research Support; Immunocore, Ltd. Funding National Institutes of Health (R01DK124267)

Anti-spastic effect induced by waggle needling correlates with KCC2-GABAA pathway in post-stroke spasticity rats

Although clinical efficacy of waggle needling has been confirmed, therapeutic mechanisms still remain poorly understood. Reduction of GABA was involved in the etiology of spasticity. Recently, accumulated evidences suggest that the inhibitory effect of GABA is determined by low intracellular chloride concentration, which is predominantly mediated by KCC2. This study was designed to investigate whether KCC2-GABAA pathway was involved in the mechanism underlying acupuncture intervention in rats with middle cerebral artery occlusion (MCAO). Three days after modeling, the rats received waggle needling, routine needling and placebo needling for 7 consecutive days. After treatment, the muscle spasticity, motor function and infarct volumes were tested. KCC2 and GABAAγ2 levels were detected via western blotting, RT-PCR and immunofluorescence. KCC2 antagonist and agonist were administered after the last intervention. We found that acupuncture, particularly waggle needling, could remarkably alleviate muscle spasticity, reverse motor deficits and reduce cerebral infraction in MCAO rats, possibly due to its effects on up-regulating expressions of KCC2 and GABAAγ2 in the cortical infarct regions. However, the effects were blocked by KCC2 antagonist. In summary, this study suggests that improvements in muscle spasticity and motor function induced by waggle needling correlates with the activation of KCC2-GABAA pathway.

GABAergic mechanisms in the suprachiasmatic nucleus that influence circadian rhythm.

The mammalian central circadian clock is located in the suprachiasmatic nucleus (SCN) of the hypothalamus. The SCN contains multiple circadian oscillators which synchronize with each other via several neurotransmitters. Importantly, an inhibitory neurotransmitter, γ-amino butyric acid (GABA), is expressed in almost all SCN neurons. In this review, we discuss how GABA influences circadian rhythms in the SCN. Excitatory and inhibitory effects of GABA may depend on intracellular Cl- concentration, in which several factors such as day-length, time of day, development, and region in the SCN may be involved. GABA also mediates oscillatory coupling of the circadian rhythms in the SCN. Recent genetic approaches reveal that GABA refines circadian output rhythms, but not circadian oscillations in the SCN. Since several efferent projections of the SCN have been suggested, GABA might work downstream of neuronal pathways from the SCN which regulate the temporal order of physiology and behavior.

Astrocytic γ-aminobutyric acid (GABA) transporters mediate guanidinoacetate transport in rat brain

Guanidinoacetate (GAA) is a biosynthetic precursor of creatine, which plays a critical role in homeostasis of high-energy phosphates in the brain, but cerebral accumulation of GAA leads to neurological complications, such as epilepsy and seizures. The purpose of the present study was to clarify the contribution of the γ-aminobutyric acid (GABA) transport systems to GAA transport in astrocytes by means of uptake studies in rat brain slices, primary astrocyte cultures and Chinese hamster ovary (CHO) cells expressing human GABA transporters (GATs). GAA uptake by rat brain slices was Na+- and Cl−-dependent, and GABA-sensitive. The inhibitory effect of GABA, a common substrate of GATs, on GAA uptake by the brain slices was similar to that of β-alanine, a selective substrate of GAT2/Slc6a13, GAT3/Slc6a11, and taurine transporter (TauT)/Slc6a6. Taurine, a high-affinity substrate of TauT/Slc6a6, exhibited a lesser inhibitory effect. In contrast, betaine, a substrate of betaine-GABA transporter 1 (BGT1)/Slc6a12, and creatine, a substrate of creatine transporter (CRT)/Slc6a8, had little inhibitory effect. A similar inhibition profile was observed in primary-cultured astrocytes. CHO cells expressing human GAT2/SLC6A13, GAT3/SLC6A11 and BGT1/SLC6A12 exhibited GAA transport, whereas CHO cells expressing GAT1/SLC6A1 did not. The Michaelis-Menten values in CHO cells expressing GAT2/SLC6A13 and GAT3/SLC6A11 were similar to those in primary-cultured astrocytes. Overall, our results suggest that astrocytic GAT2/Slc6a13 and GAT3/Slc6a11 play major roles in GAA uptake as regulatory mechanisms of GAA in rat brain, while TauT/Slc6a6, BGT1/Slc6a12, and CRT/Slc6a8 make relatively small contributions.

Detox Protocol for Acute Alcohol and Drug Withdrawal

Submit Manuscript | http://medcraveonline.com Other drugs, such as marijuana, have less intense withdrawal effects and risks due to being less addictive. While marijuana is now recognized to cause withdrawal in addicted users, it does not appear to trigger intense withdrawal requiring detox. Acute drug withdrawal produces excess stimulation of glutamate receptors, which increases the rate of nerve all firing to levels above a healthy range. Excessive glutamate excitability can produce free radicals, which can trigger a cascade of events that can lend to neuronal cell death. This cascade of events can be interrupted if the excessive glutamate activity can be decreased. Klonopin tapers stimulate gaba receptors inhibitory effects to lower the excessive glutamate receptors activity. The result is to try to restore balance between the inhibitory effects of gaba, and the excitatory effects of glutamate, to reduce the rate of nerve cell bring back towards a normal range, and prevent neuronal cell death.

Interactions between hypocretinergic and GABAergic systems in the control of activity of neurons in the cat pontine reticular formation

Anatomical studies have demonstrated that hypocretinergic and GABAergic neurons innervate cells in the nucleus pontis oralis (NPO), a nucleus responsible for the generation of active (rapid eye movement (REM)) sleep (AS) and wakefulness (W). Behavioral and electrophysiological studies have shown that hypocretinergic and GABAergic processes in the NPO are involved in the generation of AS as well as W. An increase in hypocretin in the NPO is associated with both AS and W, whereas GABA levels in the NPO are elevated during W. We therefore examined the manner in which GABA modulates NPO neuronal responses to hypocretin. We hypothesized that interactions between the hypocretinergic and GABAergic systems in the NPO play an important role in determining the occurrence of AS or W. To determine the veracity of this hypothesis, we examined the effects of the juxtacellular application of hypocretin-1 and GABA on the activity of NPO neurons, which were recorded intracellularly, in chloralose-anesthetized cats. The juxtacellular application of hypocretin-1 significantly increased the mean amplitude of spontaneous EPSPs and the frequency of discharge of NPO neurons; in contrast, the juxtacellular microinjection of GABA produced the opposite effects, i.e., there was a significant reduction in the mean amplitude of spontaneous EPSPs and a decrease in the discharge of these cells. When hypocretin-1 and GABA were applied simultaneously, the inhibitory effect of GABA on the activity of NPO neurons was reduced or completely blocked. In addition, hypocretin-1 also blocked GABAergic inhibition of EPSPs evoked by stimulation of the laterodorsal tegmental nucleus. These data indicate that hypocretin and GABA function within the context of a neuronal gate that controls the activity of AS-on neurons. Therefore, we suggest that the occurrence of either AS or W depends upon interactions between hypocretinergic and GABAergic processes as well as inputs from other sites that project to AS-on neurons in the NPO.

Valerian inhibits rat hepatocarcinogenesis by activating GABA(A) receptor-mediated signaling.

Valerian is widely used as a traditional medicine to improve the quality of sleep due to interaction of several active components with the γ-aminobutyric acid (GABA) A receptor (GABA(A)R) system. Recently, activation of GABA signaling in stem cells has been reported to suppress cell cycle progression in vivo. Furthermore, possible inhibitory effects of GABA(A)R agonists on hepatocarcinogenesis have been reported. The present study was performed to investigate modulating effects of Valerian on hepatocarcinogenesis using a medium-term rat liver bioassay. Male F344 rats were treated with one of the most powerful Valerian species (Valeriana sitchensis) at doses of 0, 50, 500 and 5000 ppm in their drinking water after initiation of hepatocarcinogenesis with diethylnitrosamine (DEN). Formation of glutathione S-transferase placental form positive (GST-P+) foci was significantly inhibited by Valerian at all applied doses compared with DEN initiation control rats. Generation of 8-hydroxy-2′-deoxyguanosine in the rat liver was significantly suppressed by all doses of Valerian, likely due to suppression of Nrf2, CYP7A1 and induction of catalase expression. Cell proliferation was significantly inhibited, while apoptosis was induced in areas of GST-P+ foci of Valerian groups associated with suppression of c-myc, Mafb, cyclin D1 and induction of p21Waf1/Cip1, p53 and Bax mRNA expression. Interestingly, expression of the GABA(A)R alpha 1 subunit was observed in GST-P+ foci of DEN control rats, with significant elevation associated with Valerian treatment. These results indicate that Valerian exhibits inhibitory effects on rat hepatocarcinogenesis by inhibiting oxidative DNA damage, suppressing cell proliferation and inducing apoptosis in GST-P+ foci by activating GABA(A)R-mediated signaling.

GABA concentration in posterior cingulate cortex predicts putamen response during resting state fMRI.

The role of neurotransmitters in the activity of resting state networks has been gaining attention and has become a field of research with magnetic resonance spectroscopy (MRS) being one of the key techniques. MRS permits the measurement of γ-aminobutyric acid (GABA) and glutamate levels, the central biochemical constituents of the excitation-inhibition balance in vivo. The inhibitory effects of GABA in the brain have been largely investigated in relation to the activity of resting state networks in functional magnetic resonance imaging (fMRI). In this study GABA concentration in the posterior cingulate cortex (PCC) was measured using single voxel spectra acquired with standard point resolved spectroscopy (PRESS) from 20 healthy male volunteers at 3 T. Resting state fMRI was consecutively measured and the values of GABA/Creatine+Phosphocreatine ratio (GABA ratio) were included in a general linear model matrix as a step of dual regression analysis in order to identify voxels whose neuroimaging metrics during rest were related to individual levels of the GABA ratio. Our data show that the connection strength of putamen to the default-mode network during resting state has a negative linear relationship with the GABA ratio measured in the PCC. These findings highlight the role of PCC and GABA in segregation of the motor input, which is an inherent condition that characterises resting state.

GABAB receptor subtypes differentially modulate synaptic inhibition in the dentate gyrus to enhance granule cell output

Activation of GABAB receptors in the dentate gyrus (DG) enhances granule cell (GC) activity by reducing synaptic inhibition imposed by hilar interneurons. This disinhibitory action facilitates signal transfer from the perforant path to the hippocampus. However, as the two main molecular subtypes, GABA(B(1a,2)) and GABA(B(1b,2)) receptors, prefer axonal terminal and dendritic compartments, respectively, they may modulate the hilar pathways at different synaptic localizations. We examined their relative expression and functions in the DG. The localization of GABAB subtypes was revealed immunohistochemically using subunit-selective antibodies in GABA(B1a)(-/-) and GABA(B1b)(-/-) mice. Effects of subtype activation by the GABAB receptor agonist, baclofen, were examined on the perforant path-stimulated GC population activities in brain slices. GABA(B(1a,2)) receptors were concentrated in the inner molecular layer, the neuropil of the hilus and hilar neurons at the border zone; while GABA(B(1b,2)) receptors dominated the outer molecular layer and hilar neurons in the deep layer, showing their differential localization on GC dendrite and in the hilus. Baclofen enhanced the GC population spike to a larger extent in the GABA(B1b)(-/-) mice, demonstrating exclusively disinhibitory roles of the GABA(B(1a,2)) receptors. Conversely, in the GABA(B1a)(-/-) mice baclofen not only enhanced but also inhibited the population spike during GABAA blockade, revealing both disinhibitory and inhibitory effects of GABA(B(1b,2)) receptors. The GABA(B(1a,2)) and GABA(B(1b,2)) receptor subtypes differentially modulate GC outputs via selective axonal terminal and dendritic locations in the hilar pathways. The GABA(B(1a,2)) receptors exclusively mediate disinhibition, thereby playing a greater role in gating signal transfer for hippocampal spatial and pattern learning.

Reversal of GABA-mediated Inhibition of the Electrically and Potassium Chloride Evoked [3H]-GABA Release from Rat Substantia Nigra Slices by DL-3-Hydroxy-3-Phenyl Pentanamide

The phenyl alcohol amides, DL-2-hydroxy-2-phenyl butyramide (CAS 52839-87-9), DL-3-hydroxy-3-phenyl pentanamide (CAS 131802-69-2, DL-HEPP) and DL-4-hydroxy-4-phenyl hexanamide (CAS 67880-30-2) and their fluorine and chlorine analogs, at a concentration of 100 micromol/L, did not displace [3H]-gamma-aminobutyric acid ([3H]-GABA, CAS 108158-36-7) from GABAA receptors and only weakly displaced [3H]-GABA and [3H]-CGP62349 (CAS 186986-97-0), a GABAB receptor antagonist, from GABAB receptors in rat brain crude synaptic membranes. The electrically and potassium chloride (15 mmol/L) evoked [3H]-GABA release in the presence of DL-HEPP, GABA and GABAB receptor ligands from rat brain substantia nigra (SN) slices was studied. R-Baclofen (CAS 69308-37-8) (10 micromol/L), a GABAB receptor agonist, produced an inhibition of the electrically evoked [3H]-GABA release and this inhibition was blocked by CGP 55845A (CAS 149184-22-5) (10 micromol/L), a GABAB receptor antagonist, but was not affected by DL-HEPP (100 micromol/L). CGP 55845A (10 micromol/L) did not alter the electrically evoked [3H]-GABA release in the absence of baclofen. The addition of DL-HEPP (100 micromol/L) alone did not affect the electrically-evoked release of [3H]-GABA release control, but it was able to significantly reduce the inhibitory effect of GABA (CAS 56-12-2) (10 micromol/L) on [3H]-GABA release evoked both by electrical and potassium chloride stimulation, in the presence of tiagabine (CAS 115103-54-3) (10 micromol/L), a GABA uptake blocker. In three preliminary experiments, bicuculline (CAS 485-49-4) (10 micromol/L) and picrotoxinin (CAS 17617-45-7) (10 micromol/L), two GABAA antagonists, inhibited the electrically evoked release of [3H]-GABA from rat SN slices, and DL-HEPP (100 micromol/L) reversed this inhibition. The mechanism of action of DL-HEPP is not known but, it might act as a negative GABA modulator in rat brain slices.

Differences in the Activation of Inhibitory Motoneuron Receptors in the Frog Rana Ridibunda by GABA and Glycine and Their Interaction

Intracellular recording of potentials was used in isolated spinal cord segments from the frog Rana ridibunda to compare the inhibitory effects of GABA and glycine on the motoneuron membrane. At equal concentrations, the response (a change in membrane potential) to application of glycine was 1.5-2 times greater than the response to GABA in terms of amplitude, and EC(50) values were 0.75 and 1.57 mM, respectively. The response to simultaneous application of GABA and glycine averaged 79.1 +/- 2.4% (n = 19) of the sum of the individual responses and 130.1 +/- 1.5% (n = 19) of the glycine response (partial occlusion). Preliminary application of glycine decreased the GABA response by 85.3 +/- 0.2% (n = 10), while preapplication of GABA decreased the glycine response by only 52.9 +/- 0.3% (n = 11). The glycine and GABA responses were specifically suppressed by strychnine and bicuculline. These results provide evidence that as in mammals, amphibian motoneurons have both glycine (predominantly) and GABA(A) receptors; they also show that asymmetrical cross inhibition can occur.

Synaptic transmission and short-term plasticity at the calyx of Held synapse revealed by multielectrode array recordings

We assessed the potential of using multielectrode arrays (MEAs) to investigate several physiological properties of the calyx of Held synapse in the medial nucleus of the trapezoid body of gerbil. Due to the large size of the synapse, it became widely employed in studies on synaptic mechanisms. Electrical stimulation at the midline evoked a characteristic compound signal consisting of a presynaptic volley ( C 1) and a postsynaptic response ( C 2). The C 1 was blocked by tetrodotoxin, whilst the C 2 was blocked by perfusion of low Ca 2+ external solution, or the AMPA-R antagonists CNQX, and GYKI52466. NMDA-R blocker D-AP5, partially inhibited the postsynaptic response at P12, but showed no effect in P30 animals. The inhibitory effects of GABA or glycine on postsynaptic responses were reciprocal with regard to animal's maturity: GABA caused a pronounced reduction of C 2 amplitude in P20–22 animals, while glycine showed a stronger inhibition in P27–28 animals. Low-frequency super-threshold stimulation of the afferents induced facilitation of the postsynaptic C 2 amplitudes and only minor changes in temporal characteristics of the signals. At stimulation frequencies >200 Hz, however, significant depression occurs accompanied by increases in transmission delay and in the width of the postsynaptic response. This study suggests MEAs as a useful tool to study calyx of Held synapse by simultaneous recordings of pre- and postsynaptic elements of synaptically interconnected neurons in the auditory brainstem. Moreover, MEAs enable convenient analysis of activity-dependent depression and modulation of neuronal activity by glycine and GABA at later developmental stages not accessible to patch recordings.

Human peripheral blood mononuclear cells express GABA A receptor subunits

The polymerase chain reaction was used to screen human peripheral blood mononuclear cells (PBMC) and Jurkat cells for the presence of GABA A receptor subunit mRNAs. Positive signals were detected for the α1, α3, β2, β3, δ and ɛ subunit mRNAs in both cell populations, with the Jurkat cells giving a positive signal for some additional species. Real-time PCR was used to confirm that PBMC, lymphocytes and monocytes contained significant levels of the α1 subunit mRNA and that PBMC and lymphocytes contained low levels of β2 mRNA. The α1 subunit was detected in PBMC and fractionated T-cell populations, as well as Jurkat and HL-60 cell lines, by Western blotting and immunofluorescence using a specific antibody. The application of 1 mM GABA reduced the specific increase in intracellular PBMC Ca 2+ levels produced by addition of 1 nM fMLP: this effect was mimicked by muscimol, but not glycine, and was blocked by bicuculline. The inhibitory effect of GABA was limited to a subset of PBMC. We conclude that cells within the human PBMC population, including lymphocytes, express functional GABA A receptors and these receptors may modulate immune responses.

Effects of GABA and bicuculline on the electrical activity of rat olfactory placode neurons derived at E13.5 and cultured for 1 week on multi-electrode dishes.

The present study was performed to record the electrical activity of olfactory placode neurons and to check the effect of GABA and bicuculline on it. Olfactory placodes obtained at day 13.5 of gestation were cultured for 1 week on multi-electrode dishes. Olfactory placode neurons showed spontaneous firing, with firing rates of 0.77 +/- 0.05 Hz (0.03-3.82 Hz, n = 12), but there was no bursting activity. Perfusion with 10 microM GABA almost immediately inhibited 8 of 11 firing activities (we could not test it in 1 activity). In contrast, perfusion with 10 microM bicuculline induced facilitation in 5 of 12 activities and did not induce any change in 7 other activities. Statistical analysis by chi(2)-test showed a significant difference in the response of neurons to the two drugs. Fisher's exact probability test showed that the inhibitory effect of GABA was significant (p<0.05) whereas neither the facilitatory effect nor the lack of effect of bicuculline was significant (p>0.1). These results suggest that cultured olfactory placode neurons, even in a probably immature stage, respond to GABA with inhibition, as generally observed at mature stages.

The subcellular localization of GABA(B) receptor subunits in the rat substantia nigra.

The inhibitory effects of GABA within the substantia nigra (SN) are mediated in part by metabotropic GABA(B) receptors. To better understand the mechanisms underlying these effects, we have examined the subcellular localization of the GABA(B) receptor subunits, GABA(B1) and GABA(B2), in SN neurons and afferents using pre-embedding immunocytochemistry combined with anterograde or retrograde labelling. In both the SN pars compacta (SNc) and pars reticulata (SNr), GABA(B1) and GABA(B2) showed overlapping, but distinct, patterns of immunolabelling. GABA(B1) was more strongly expressed by putative dopaminergic neurons in the SNc than by SNr projection neurons, whereas GABA(B2) was mainly expressed in the neuropil of both regions. Immunogold labelling for GABA(B1) and GABA(B2) was localized in presynaptic and postsynaptic elements throughout the SN. The majority of labelling was intracellular or was associated with extrasynaptic sites on the plasma membrane. In addition, labelling for both subunits was found on the presynaptic and postsynaptic membranes at symmetric, putative GABAergic synapses, including those formed by anterogradely labelled striatonigral and pallidonigral terminals. Labelling was also observed on the presynaptic membrane and at the edge of the postsynaptic density at asymmetric, putative excitatory synapses. Double immunolabelling, using the vesicular glutamate transporter 2, revealed the glutamatergic nature of many of the immunogold-labelled asymmetric synapses. The widespread distribution of GABA(B) subunits in the SNc and SNr suggests that GABA(B)-mediated effects in these regions are likely to be more complex than previously described, involving presynaptic autoreceptors and heteroreceptors, and postsynaptic receptors on different populations of SN neurons.

Role of GABA A and GABA B receptors in GABA-induced inhibition of rat red nucleus neurons

We investigated GABA receptor subtypes mediating GABA-induced inhibition of red nucleus (RN) neuronal firing recorded extracellularly from anaesthetized rats. GABA response was mimicked by the GABA A agonists muscimol and isoguvacine in all cases and was partially blocked by the GABA A antagonist bicuculline. The GABA B agonist baclofen induced a long-lasting inhibition in 84% of cells. Neurons responding to either GABA A or GABA B agonists were equally distributed within the RN. The GABA C receptor agonist cis-amino-crotonic acid (CACA) did not modify RN neuronal firing; at high doses CACA occasionally induced inhibition abolished by bicuculline and thus mediated by GABA A receptors. We conclude that the inhibitory effects of GABA in the RN are mediated by both GABA A and GABA B receptors, whereas GABA C receptors are not involved.

Involvement of GABA(B) receptors in presynaptic inhibition of fibers of the descending projections of the spinal cord in the frog Rana ridibunda.

Isolated spinal cord preparations from Rana ridibunda frogs were used for studies of the effects of the GABA(B) receptor agonists (-)-baclofen (50 and 100 microM) and GABA (4-8 mM) and the specific GABA(B) receptor antagonist 2-hydroxysaclofen (100 microM) on the transmission of signals from fibers of the ventral columns monosynaptically connected with motoneurons in segments 9 and 10. These experiments showed that (-)-baclofen (50 and 100 microM) produced significant and dose-dependent suppression of excitatory postsynaptic potentials (EPSP) in motoneurons and ventral root potentials evoked by stimulation of fibers of the ipsi- and contralateral ventral columns. The inhibitory effect of (-)-baclofen (100 microM) on descending EPSP was 35-50% blocked by the GABA(B) receptor antagonist 2-hydroxysaclofen (0.2 mM). The inhibitory effect of GABA (4-8 mM) on descending EPSP was 60% blocked by the GABA(A) receptor antagonist picrotoxin (0.05 mM). (-)-Baclofen (50 microM) and GABA (4 and 6 mM) were found to have inhibitory effects on ventral root potentials evoked by stimulation of the ipsi- and contralateral ventral columns. The data obtained here, as well as data obtained by pharmacological analysis and conditioning by stimulation of the ipsi- and contralateral ventral columns, are regarded as a significant argument supporting the existence of GABA(B) receptor-mediated presynaptic inhibition of descending fibers connected monosynaptically to spinal cord motoneurons in the frog Rana ridibunda.

Inhibitory effects of GABA B receptor agonists on swallowing in the dog

The effects of the GABA B receptor agonists baclofen (1.4 and 7 μmol/kg i.v.) and CGP 44532 ([(2 S)-3-amino-2-hydroxypropyl]methyl phosphinic acid], 0.2 and 0.7 μmol/kg i.v.) on transient lower esophageal sphincter relaxations and spontaneous and pharyngeally stimulated swallowing were investigated in conscious dogs. Both compounds inhibited transient lower esophageal sphincter relaxations dose-dependently, CGP 44532 being approximately fivefold more potent. In experiments designed to measure transient lower esophageal sphincter relaxations, spontaneous swallowing was suppressed by both compounds. When swallowing was evoked by intrapharyngeal water injection, both baclofen and CGP 44532 reduced the occurrence of primary peristalsis. It is concluded that centrally acting GABA B receptor agonists inhibit spontaneous and stimulated swallowing probably through an action in the central pattern generator for swallowing.

Mechanism underlying γ-aminobutyric acid-induced antihypertensive effect in spontaneously hypertensive rats

We examined the effects of γ-aminobutyric acid (GABA) on the blood pressure in spontaneously hypertensive rats and normotensive Wistar–Kyoto rats. A single oral administration (0.5 mg/kg) significantly lowered the systolic blood pressure in spontaneously hypertensive rats, but not in normotensive rats. In the mesenteric arterial bed, the perivascular nerve stimulation-induced increase in perfusion pressure and noradrenaline release were significantly inhibited by GABA in spontaneously hypertensive rats, but not in normotensive rats, and attenuated by the selective GABA B receptor agonist, baclofen, but not by the selective GABA A receptor agonist muscimol. The inhibitory effects of GABA on the perivascular nerve stimulation-induced increase in perfusion pressure and noradrenaline release were completely antagonized by the selective GABA B receptor antagonist, saclofen, but not by the selective GABA A receptor antagonist, bicuculline. These results suggest that, in spontaneously hypertensive rats, GABA has an antihypertensive effect due to its inhibition of noradrenaline release from sympathetic nerves in the mesenteric arterial bed via presynaptic GABA B receptors.

The study of the role of the GABA A and GABA B receptors in presynaptic inhibition of primary afferents of spinal cord of the frog Rana ridibunda

The role of GABAA- and GABAB-receptors in presynaptic inhibition of primary afferent fibers was studied on an isolated preparation of the spinal cord of the frog Rana ridibunda. It is shown that the inhibitory effect of GABA on synaptic transmission from afferent fiber to motoneuron is caused by activation of both GABAA- and GABAB-receptors. A temporal correlation (± 5 min) was shown between the blocking action of bicuculline (a specific antagonist of GABAA-receptors) on primary afferent fiber depolarization (PAD) and its potentiating effect on the excitatory postsynaptic potential (EPSP) at parallel intracellular recording of EPSP in motoneuron and PAD in axons of the dorsal root. As a basis of this correlation, the single GABAA-receptor mechanism is discussed, which mediates the effect of bicuculline on PAD and EPSP. When a specific agonist of GABAB-receptor, baclofen, and an antagonist of GABAB-receptor, 2(OH)-saclofen, were applied, the obtained data indicated an involvement of GABAB-receptors in inhibition of synaptic transmission from afferent fibers to the motoneuron. Analysis of parameters of the unitary synaptic responses recorded in the control experiments and of their changes under the effect of (− )-baclofen indicates that the inhibitory action caused by activation of GABAB-receptors develops at the presynaptic level.