Inhibitory Effect Of GABA Research Articles

The effects of ionotropic agonists of excitatory amino acids on the release of arginine vasopressin in rat hypothalamic slices.

The effects of ionotropic excitatory amino acids agonists on the release of vasopressin from rat hypothalamic slices were studied. Incubation with increasing doses of NMDA, kainate or AMPA decreased the release of vasopressin in a dose-dependent manner. The values of the IC50 were 1.0, 9.6, or 3.7 x 10-8 M, respectively. The inhibitory effect of the various excitatory amino acids tested was blocked by coincubation with their respective antagonists. Vasopressin secretion was stimulated to 140.3 +/- 7.6% of controls when the slices were obtained from chronically (7 days) salt-loaded rats. Addition of 1 x 10-7 M NMDA or 1 x 10-6 M kainate to the incubation medium antagonized the salt loading-induced increase in vasopressin release. Incubation with 1 x 10-4 M tetrodotoxin did not change basal vasopressin release, but it blocked the decrease in vasopressin secretion induced by 1 x 10-7 M NMDA or 1 x 10-6 M kainate or 1 x 10-6 M AMPA. Incubation with 1 x 10-5 M phaclophen (a GABAB antagonist) and 1 x 10-5 M bicuculline (a GABAA antagonist) was without effect on basal vasopressin secretion while it reversed the inhibition of vasopressin release induced by 1 x 10-7 M NMDA. Incubation with 1 x 10-6 M GABA alone decreased vasopressin secretion to 64.6 +/- 6.9% of control values. The inhibitory effect of GABA did not change when 1 x 10-7 M NMDA was added to the incubation medium. These findings demonstrate that ionotropic excitatory amino acids agonists inhibit vasopressin secretion from hypothalamic slices. They strongly suggest that this inhibitory effect is mediated through local GABAergic interneurones.

GABA A and GABA B receptors in the anterior piriform cortex modulate feeding in rats

The effects of GABA A and GABA B receptors in the anterior piriform cortex (APC) on intake of an amino acid imbalanced diet and a basal diet were evaluated in rats. Administration of muscimol (GABA A receptor agonist) to the APC immediately suppressed ingestion of both amino acid imbalanced and basal diets. Central administration of bicuculline (a GABA A receptor antagonist) stimulated feeding of the amino acid imbalanced diet but had no effect on intake of the basal diet. The GABA B receptor antagonist phaclofen decreased consumption of the basal diet but did not affect consumption of the amino acid imbalanced diet. These findings demonstrate that manipulation of GABA-sensitive cells in the APC can have a pronounced effect on feeding behavior that is not selective to aminoprivic feeding. However, these data suggest that GABA A and GABA B receptors may function as regulators that are activated by monoaminergic systems and neuropeptides in response to amino acid imbalanced diet intake. Inhibitory effects of GABA A and GABA B receptors may modulate the pyramidal cells, contributing to the reduced feeding response to the amino acid imbalanced diet. Also, transcription of mRNA for both GABA receptors and the GABA reuptake transporter was affected by a threonine deficient but not a corrected diet, compared to the basal diet. Taken together, these results support the involvement of GABA receptors in the APC in feeding in general and the responses to amino acid deprivation in vivo.

Involvement of GABAergic Systems in Manifestation of Pharmacological Activity of Desipramine

We have conducted this study to elucidate the influence of GABAergic systems on manifestation of pharmacological activity of desipramine using both pharmacological and electrophysiological methods. Desipramine (20 mg/kg, i.p.) significantly blocked the adjuvant-induced thermal hyperalgesia, which was facilitated by treatment with the GABA(A) antagonist picrotoxin (2 mg/kg, i.p.) or the GABA(B) antagonist saclofen (2 mg/kg, i.p.). This analgesic effect of desipramine was antagonized by post-treatment with picrotoxin or saclofen. However, none of these compounds showed any effect in normal animals without adjuvant-induced inflammation. In a slice preparation of the hippocampus, treatment with GABA (10(-5)-5 x 10(-4) M), baclofen (10(-5)-10(-4) M) or muscimol (10(-5)-10(-4) M) inhibited the field potential evoked in pyramidal neurons by Schaffer collateral stimulation. The inhibitory effect of GABA was facilitated by concurrent application of desipramine, carbamazepine or diazepam at a concentration of 5 x 10(-5)-2 x 10(-4) M. The rank of order of facilitation is: desipramine > carbamazepine > diazepam. Desipramine also enhanced the inhibitory effect of baclofen and muscimol. These results suggest that desipramine causes GABAergic systems to activate still more, and this phenomenon appears to be involved in manifestation of the pharmacological activity of desipramine such as antinociception.

Effect of experimental diabetes on GABA-mediated inhibition of neurally induced contractions in rat isolated trachea.

1. In the present study, we investigated the effect of GABA and selective GABA agonists and antagonists on neurally induced tracheal contractions in streptozotocin (STZ) diabetic rats. 2. Contractile responses to electrical field stimulation (EFS) in rat tracheal rings were completely abolished by atropine and tetrodotoxin, but were unaffected by the ganglion blocker hexamethonium, indicating that they were mediated via neuronal release of acetylcholine (ACh). 3. Contractions induced by EFS, but not by exogenous ACh, were inhibited by GABA and the selective GABA(B) receptor agonist baclofen, but not by the selective GABA(A) receptor agonist 3-aminopropane sulphonic acid. The inhibitory effects of GABA or baclofen were not affected by the GABA(A) antagonist bicuculline, but were significantly reversed by the GABA(B) antagonist phaclofen. 4. The inhibitory effects of both GABA and baclofen were found to be significantly greater in trachea from control rats compared with tissues from diabetic rats. 5. Non-adrenergic, non-cholinergic relaxation responses elicited by EFS in precontracted tracheal rings from diabetic and control rats were similar in magnitude and were unaffected by GABA or GABA analogues. 6. These results suggest that GABA decreases the response to EFS by directly inhibiting the evoked release of ACh through GABA(B) receptors in rat trachea and that STZ-induced diabetes causes an impairment in the inhibitory effect of GABA on neurally induced contractions in this tissue.

The effects of acute ethanol exposure on inhibitors of hepatic regenerative activity in the rat.

The purpose of this study was to identify the mechanism(s) whereby acute ethanol exposure inhibits hepatic regenerative activity in the rat. Adult, male, Sprague-Dawley rats (200-250 g) were randomized to receive either ethanol (1 g/kg i.p. q 4 h) or an equal volume of saline (controls) for 24 h beginning 1 h prior to a 70% partial hepatectomy (PHx). At 0, 3, 6, 12 and 24 h post-PHx, rats were sacrificed (N = 4-6/group), and the expression of the following genes associated with inhibition of hepatocyte proliferation were documented; p53, p21, transforming growth factor-beta1 (TGF-beta1) and gamma aminobutyric acid transport protein (GABA-TP). Inhibition of hepatic regenerative activity was confirmed by 3H-thymidine incorporation into hepatic DNA at 24 h post-PHx. The results of the study revealed that in ethanol-treated rats, DNA synthesis was inhibited by 37% when compared to saline-treated controls (p < 0.01). Regarding suppressor gene expression, both p21 and TGF-beta1 mRNA expression in ethanol-treated rats were similar to those obtained in saline-treated controls. Although p53 mRNA expression differed in the two groups, in the ethanol-treated group, p53 mRNA expression was decreased rather than increased (relative to controls) at 24 h post-PHx, a finding not in keeping with inhibition of DNA synthesis. GABA-TP mRNA was strongly expressed prior to PHx in both ethanol- and saline-treated rats. Following PHx, GABA-TP mRNA expression decreased in both groups but remained low in the saline-treated group while returning to pre-PHx values in ethanol-treated rats. In summary, the results of this study indicate that the inhibitory effects of ethanol on hepatic regeneration are not associated with significant or the appropriate changes in mRNA expression of the p53, p21 or TGF-beta1 suppressor genes. On the other hand, transcriptional changes in GABA-TP gene expression post-PHx are in keeping with an inhibitory effect of GABA on hepatic regeneration.

Role of Potassium Channels in the GABA Inhibitory Action on the Cholinergic Response to Electrical Field Stimulation in Guinea-pig Ileum

γ-Aminobutyric acid B (GABAB) receptor activation inhibits the cholinergic response to electrical field stimulation (EFS) in guinea-pig ileum. The involvement of potassium channels in this GABA inhibitory action was investigated. The modulatory effect of GABA was not modified after pre-incubation of the organ with 1 mM tetraethylammonium chloride, a non-selective potassium channel antagonist. Nevertheless, the non-selective K+ channels antagonists, 4-aminopyridine (3 mM) and quinine (200 μM), and the selective KATP channel antagonist glibenclamide (1 μM), significantly reduced the inhibitory effects of GABA. The results suggest a coupling between GABAB receptors and KATP channels in the guinea-pig ileum.

N-Type Calcium Channels and Their Regulation by GABABReceptors in Axons of Neonatal Rat Optic Nerve

Axons of neonatal rat optic nerves exhibit fast calcium transients in response to brief action potential stimulation. In response to one to four closely spaced action potentials, evoked calcium transients showed a fast-rising phase followed by a decay with a time constant of approximately 2-3 sec. By selective staining of axons or glial cells with calcium dyes, it was shown that the evoked calcium transient originated from axons. The calcium transient was caused by influx because it was eliminated when bath calcium was removed. Pharmacological profile studies with calcium channel subtype-specific peptides suggested that 58% of the evoked calcium influx was accounted for by N-type calcium channels, whereas L- and P/Q-type calcium channels had little, if any, contribution. The identity of the residual calcium influx remains unclear. GABA application caused a dramatic reduction of the amplitude of the action potential and the associated calcium influx. When GABAA receptors were blocked by bicuculline, the inhibitory effect of GABA on the action potential was eliminated, whereas that on the calcium influx was not, indicating involvement of GABAB receptors. Indeed, the calcium influx was inhibited by the GABAB receptor agonist baclofen. This baclofen effect was occluded by a previous block of N-type calcium channels and was unaffected by the broad-spectrum K+ channel blocker 4-AP. We conclude that neonatal rat optic nerve axons express N-type calcium channels, which are subjected to regulation by G-protein-coupled GABAB receptors. We suggest that receptor-mediated inhibition of axonal calcium channels plays a protective role in neonatal anoxic and/or ischemic injury.

GABA Exerts Stimulatory and Inhibitory Influences on Gonadotropin II Secretion in the Atlantic Croaker (Micropogonias undulatus)

Possible involvement of γ-aminobutyric acid (GABA) in the control of gonadotropin II (GTH II) secretion was investigated during different phases of the seasonal gonadal cycle of the Atlantic croaker (Micropogonias undulatus). Intraperitoneal administration of GABA (100 µg/g) elicited a significant increase in plasma GTH II levels in croaker with regressed or pre-recrudescence phase gonads, whereas it did not significantly affect GTH II levels during the early- to mid-recrudescence phase of the gonadal cycle in this species. On the other hand, GABA induced a dose-related inhibition of GTH II secretion in fish with fully recrudesced gonads. Similar opposite changes in GTH II levels were observed when croaker with regressed and fully recrudesced gonads were injected with the GABA transaminase inhibitor, γ-vinyl GABA (10 and 100 µg/g). This suggests an involvement of endogenous GABA in the control of GTH II secretion. GABA (1–100 µM) did not significantly alter GTH II release from pituitary fragments from regressed or fully recrudesced fish incubated in vitro, suggesting the lack of a major direct influence at the pituitary level. Muscimol (GABA_A receptor agonist, 1 µg/g) treatment mimicked both the stimulatory and inhibitory effects of GABA, whereas injection of baclofen (GABA_B receptor agonist, 1 µg/g) had no significant effect on basal or a gonadotropin-releasing hormone (GnRH) analog-induced GTH II secretion. Pretreatment of fish with bicuculline (1 µg/g), a GABA_A antagonist, completely blocked the stimulatory influence of GABA on GTH II secretion in croaker with regressed gonads and partially blocked the inhibitory effect in individuals with fully recrudesced gonads. The results demonstrate that GABA exerts opposite influences on GTH II secretion at two different stages of the gonadal cycle of croaker, and acts primarily via GABA_A type receptors. Thus, GABAergic pathways may be important in mediating seasonal influences on GTH II secretion in this species.

GABA inhibits endozepine release from cultured rat astrocytes

In the mammalian brain, the endogenous ligands for benzodiazepine receptors (also called endozepines) are predominantly synthesized by glial cells. It has recently been reported that rat astrocytes in primary culture release substantial amounts of endozepines. The aim of the present study was to investigate the possible involvement of GABA in the control of endozepine release. Exposure of cultured rat astrocytes to GABA (10−7 to 10−5 M) induced a dose-related inhibition of endozepine secretion. At higher doses (3 × 10−5 to 10−3 M), the effect of GABA gradually diminished. The inhibitory effect of GABA (10−5 M) was mimicked by the GABAB receptor agonist baclofen (10−5 M). In contrast, the GABAA receptor agonists 3APS and isoguvacine (10−5 M each) did not modify endozepine release. The inhibition of endozepine secretion evoked by GABA and baclofen (10−5 M each) was totally abrogated by the specific GABAB receptor antagonist phaclofen (10−4 M). GABA and baclofen caused a significant inhibition of forskolin-evoked production of cAMP in astrocytes and this effect was abolished in the presence of phaclofen. In contrast, isoguvacine had no effect on cAMP production. Exposure of astrocytes to dbcAMP induced a time- and dose-dependent stimulation of endozepine release. These data indicate that GABA, acting through GABAB receptors negatively coupled to adenylyl cyclase, inhibits endozepine release from cultured rat astrocytes. The secretion of endozepines thus appears to be a valuable marker to monitor astrocyte activity. GLIA 25:404–411, 1999. © 1999 Wiley-Liss, Inc.

GABA inhibits endozepine release from cultured rat astrocytes

In the mammalian brain, the endogenous ligands for benzodiazepine receptors (also called endozepines) are predominantly synthesized by glial cells. It has recently been reported that rat astrocytes in primary culture release substantial amounts of endozepines. The aim of the present study was to investigate the possible involvement of GABA in the control of endozepine release. Exposure of cultured rat astrocytes to GABA (10(-7) to 10(-5) M) induced a dose-related inhibition of endozepine secretion. At higher doses (3 x 10(-5) to 10(-3) M), the effect of GABA gradually diminished. The inhibitory effect of GABA (10(-5) M) was mimicked by the GABA(B) receptor agonist baclofen (10(-5) M). In contrast, the GABA(A) receptor agonists 3APS and isoguvacine (10(-5) M each) did not modify endozepine release. The inhibition of endozepine secretion evoked by GABA and baclofen (10(-5) M each) was totally abrogated by the specific GABA(B) receptor antagonist phaclofen (10(-4) M). GABA and baclofen caused a significant inhibition of forskolin-evoked production of cAMP in astrocytes and this effect was abolished in the presence of phaclofen. In contrast, isoguvacine had no effect on cAMP production. Exposure of astrocytes to dbcAMP induced a time- and dose-dependent stimulation of endozepine release. These data indicate that GABA, acting through GABA(B) receptors negatively coupled to adenylyl cyclase, inhibits endozepine release from cultured rat astrocytes. The secretion of endozepines thus appears to be a valuable marker to monitor astrocyte activity.

Role of arachidonic acid in the regulation of the NMDA-evoked release of acetylcholine in striatal compartments.

The role of endogenously released arachidonic acid in the control of the NMDA (50 microM)-evoked release of [3H]-acetylcholine previously formed from [3H]-choline was investigated in striosome-enriched areas and in the matrix of the rat striatum using a microsuperfusion procedure in vitro. Experiments were performed with either mepacrine (0.2 microM) or bovine serum albumin (BSA, 0.02%) which inhibits phospholipase A2 activity or binds endogenously released arachidonic acid, respectively. Both treatments similarly reduce the NMDA-evoked release of [3H]-acetylcholine, this effect being more pronounced in striosomes than in the matrix. These reductions result from a facilitation of dopamine release, since they were not observed in the presence of (-)sulpiride, the D2 dopamine receptor antagonist. Moreover, the superfusion with BSA was shown to enhance the release of [3H]-dopamine (formed from [3H]-tyrosine), this effect being of larger amplitude in striosomes than in the matrix. In control conditions, due to the blockade of the presynaptic inhibitory effect of GABA on dopamine release, bicuculline (GABA(A) receptor antagonist) reduces the NMDA-evoked release of [3H]-acetylcholine in both striatal compartments. Bicuculline was no longer effective following superfusions with either mepacrine or BSA, suggesting that these treatments eliminate the GABAergic presynaptic inhibitory control on dopamine transmission and thus lead to the dopamine-mediated inhibition of [3H]-acetylcholine release. These results indicate that arachidonic acid endogenously formed under weak stimulation of NMDA receptors contributes to the regulation of the evoked release of [3H]-acetylcholine by facilitating GABAergic transmission and that this process is more important in striosomes than in the matrix.

Calcium currents and GABAB receptors in the dorsal sensory cells of the lamprey spinal cord.

Patch-clamp studies were performed on the isolated dorsal sensory cells of the spinal cords of three species of lamprey, Ichthyomyzon unicuspis, Petromyzon marinus, and Lampetra fluviatilis, to measure changes in the amplitudes of calcium current induced by GABA and its specific antagonists and agonists. The experiments showed that GABA (4 mM) reduced the peak amplitude of the calcium current by 28.5 +/- 4.9%, with subsequent recovery to 96.2 +/- 9.2% of control (n = 45). The GABAB agonist baclofen had similar effects. The GABAA agonists glycine and taurine had no effect on the Ca2+ current. The inhibitory effect of GABA was blocked by 2-hydroxysaclofen (a GABAB antagonist), but persisted in the presence of bicuculline (a GABAA antagonist). These results are evidence that the membranes of dorsal sensory cells contain GABAB receptors, which significantly increases our understanding of the mechanisms of presynaptic inhibition in the spinal cords of the cyclostomata.

Clozapine's antipsychotic effects do not depend on blockade of 5-HT3 receptors.

Sixteen known 5-HT3 receptor blockers, including clozapine, fully or partially reverse the inhibitory effect of 1 microM GABA on [35S]TBPS binding, indicating that they are also GABA(A) antagonists, some of them selective for subsets of GABA(A) receptors. The 5-HT3 receptor blocker, ondansetron, has been reported to produce some antipsychotic and anxiolytic effects. However, no antipsychotic effects have been reported for a large number of highly potent 5-HT3 receptor blockers. Like clozapine, ondansetron partially reverses the inhibitory effect of GABA on [35S]TBPS binding. Additivity experiments suggest that ten 5-HT3 receptor blockers tested at low concentrations preferentially block subtypes of GABA(A) receptors that are among those blocked by clozapine. Wiley and Porter (29) reported that MDL-72222, the most potent GABA(A) antagonist described here, partially generalizes (71%) with clozapine in rats trained to discriminate an interoceptive clozapine stimulus, but only at a dose that severely decreases responding. Tropisetron (ICS-205,930) exhibits both GABA-positive and GABA-negative effects. R-(+)-zacopride is 6-fold more potent than S-(-)-zacopride as a GABA(A) antagonist. We conclude that the observed antipsychotic and, possibly, anxiolytic effects of some 5-HT3 receptor blockers are due to selective antagonism of certain GABA(A) receptors, and not to blockade of 5-HT3 receptors. We speculate that the anxiolytic and sedative effects of clozapine and several other antipsychotic drugs may be due to selective blockade of alpha1beta2gamma2 GABA(A) receptors which are preferentially located on certain types of GABAergic interneurons (probably parvalbumin positive). Blockade of these receptors will increase the inhibitory output of these interneurons. So far, no highly potent GABA(A) antagonists with clozapine-like selectivity have been identified. Such compounds may exhibit improved clozapine-like antipsychotic activity.

Both GABA A and GABA B receptors participate in suppression of [Ca 2+] i pulsing in toad melanotrophs

The receptor mechanisms involved in the inhibitory effect of γ-aminobutyric acid (GABA) in suppressing spontaneous [Ca 2+] i pulsing in melanotrophs of Xenopus laevis were investigated. The selective GABA B receptor agonist, baclofen reversibly arrested [Ca 2+] i pulsing. This inhibition was unaffected by the selective GABA A receptor antagonist, bicuculline methiodide, but was blocked by the selective GABA B receptor antagonist, CGP 35348 (3-aminopropyl diethyoxymethyl phosphinic acid). The selective GABA A receptor agonist, muscimol, also arrested [Ca 2+] i pulsing after causing a transient rise in [Ca 2+] i. This biphasic response to muscimol was unaffected by CGP 35348, but was blocked by bicuculline. The inhibitory effect of GABA was unaffected by either CGP 35348 or bicuculline when given alone, but was blocked by both antagonists given together. In cells pretreated with pertussis toxin, the response to baclofen was completely lost, whereas responses to GABA and muscimol persisted; the response to GABA was blocked by bicuculline alone. Thus, both GABA A and GABA B receptors are involved in the inhibitory effect of GABA in suppressing spontaneous [Ca 2+] i pulsing in Xenopus melanotrophs.

Clozapine and some other antipsychotic drugs may preferentially block the same subset of GABA(A) receptors.

Selective blockade of a subset of GABA(A) receptors may be involved in the antipsychotic effects of Clozapine and several other antipsychotic drugs. Seven antipsychotic drugs, and 11 drugs classified as antidepressants that only partially reverse the inhibitory effect of 1 microM GABA on [35S]TBPS binding, do not yield additive reversal when tested pairwise with Clozapine, which also only partially reverses the inhibitory effect of GABA. This suggests that all of these antipsychotic/antidepressant drugs may block a common subset of GABA(A) receptors. DMCM and Ro 5-4864 are also partial reversers of GABA's inhibitory effect, but they yield additive reversals when tested pairwise with the antipsychotic/antidepressant drugs, and also with each other, suggesting that DMCM, Ro 5-4864, and the antipsychotic drugs define three heterogeneous subsets of GABA(A) receptors, with variable overlap, depending on the drug. Several potent ligands for benzodiazepine binding sites can block the GABA inhibitory effects of DMCM and Ro 5-4864, but with different patterns: the ligands generally blocked DMCM less potently, but more completely than Ro 5-4864. Ro 5-4864 was not blocked by Flumazenil or CGS-8216, ligands that potently blocked DMCM. Nine additional antipsychotic/antidepressant drugs, as well as Clozapine, and 7 "classical" GABA(A) receptor blockers, all of which reversed GABA nearly completely, when tested at lower concentrations that only reverse approximately 20-35%, yielded almost complete additivity when tested pairwise with DMCM or Ro 54864. Another convulsant benzodiazepine, KW-1937, a positional isomer of Brotizolam, fully reverses the inhibitory effect of 1 microM GABA. At a lower concentration yielding about 50% reversal, KW-1937 is completely additive with DMCM, but entirely nonadditive with Ro 5-4864. The 50% reversal obtained with KW-1937 was potently blocked by Triazolam, but with a plateau similar to that obtained with Ro 5-4864. The results with KW- 1937 suggest that its 50% reversal largely corresponds to the reversal obtained with Ro 5-4864, and that virtually all of the [35S]TBPS binding sites inhibited by 1 microM GABA are coupled to benzodiazepine binding sites. The fraction of GABA(A) receptors preferentially blocked by all the antipsychotic/antidepressant drugs, roughly 25% of the [35S]TBPS binding sites inhibited with 1 microM GABA, are sensitive to KW-1937, but not to DMCM or to Ro 5-4864.

Role of nitric oxide/cyclic GMP pathway in the inhibitory effect of GABA and dopamine on prolactin release.

The anterior pituitary gland is a site of nitric oxide (NO) production and action, suggesting a local regulatory function. We recently reported that NO inhibits in vitro prolactin release. The aim of the present study was to establish the mechanism of action of NO on prolactin release and to determine whether NO is involved in the inhibitory effect of GABA on prolactin release. Since NO exerts its action through cGMP by activating guanylate cyclase in different tissues, we examined the effect of sodium nitroprusside (NP), a NO releaser, on intrapituitary cGMP levels. Incubation of anterior pituitary glands with 0.5 mM NP 4-fold increased intrapituitary cGMP content, but decreased intrapituitary cAMP levels. In addition, we studied the effect of NP on prolactin release in the presence of LY 83583, an inhibitor of guanylate cyclase activity and 3-isobutyl-1-methylxanthine (IBMX), an inhibitor of phosphodiesterase activity. 10 microM LY 83583 and 0.5 mM IBMX blocked the inhibitory effect of NP on prolactin release. (10(-3) M) 8Br-cGMP, an analogue of cGMP, mimicked the effect of NP on prolactin release. On the other hand, NO seems to be involved in the inhibitory effect of GABA on prolactin release since hemoglobin, a scavenger of NO, and Nw-nitro-L-arginine methyl ester, an inhibitor of NO synthase (NOS), blocked the pituitary response to GABA. Moreover, GABA (10(-6) M) stimulated NOS activity by almost 50%. GABA increased intrapituitary cGMP levels and decreased cAMP. Dopamine stimulated NOS activity weakly. These observations suggest that NO, acting through the guanylate cyclase-cGMP pathway, inhibits prolactin secretion. In addition, NO may be involved in the inhibitory effect of GABA and dopamine on prolactin release.

Multiple GABA receptor subtypes mediate inhibition of calcium influx at rat retinal bipolar cell terminals

Inhibitory effects of GABA on K(+)-evoked Ca2+ influx into rat retinal bipolar cell terminals were studied using calcium imaging methods. Application of high K+ evokes a sustained, reversible increase in [Ca2+]i at bipolar cell terminals, which occurs mainly via dihydropyridine-sensitive (L-type) Ca2+ channels. There are at least two GABA receptor subtypes coexisting at bipolar cell terminals: a conventional GABAA receptor and a bicuculline/baclofen-insensitive GABA receptor. Activation of either GABA receptor inhibited the K(+)-evoked Ca2+ response. However, these two GABA receptor subtypes have distinct properties. GABAA receptors suppress the Ca2+ response only at relatively high concentrations of agonist, and with fas kinetics and a narrow dynamic range. In contrast, the bicuculline/baclofen-insensitive GABA receptors produce inhibition on the Ca2+ response at a much lower concentration of agonist, and with slow onset and a wider dynamic range. The pharmacologic profile of the bicuculline/baclofen-insensitive GABA receptor at bipolar cell terminals is most similar to the GABAC receptor reported by Feigenspan et al. (1993). Unlike the GABAC receptors described in other species, it is extremely insensitive to picrotoxin. Therefore, it may be appropriate to refer to this receptor as a picrotoxin-insensitive GABAc receptor. 3-Aminopropyl(methyl)phosphinic acid (3-APMPA) and 3-aminopropylphosphonic (3-APA), two phosphate analogs of GABA, selectively antagonize the picrotoxin-insensitive GABAc receptors but not the GABAA receptors in this system. These results imply a functional role for multiple GABA receptors in regulating synaptic transmission at bipolar cell terminals.

Long‐Term GABA Treatment Elicits Supersensitivity of Quisqualate‐Preferring Metabotropic Glutamate Receptor in Cultured Rat Cerebellar Neurons

In primary cultures of rat cerebellar granule neurons, GABA treatment (50 microM, 7 days) caused a withdrawal supersensitivity selective for the metabotropic glutamate receptors that mainly prefer L-glutamate, quisqualate and, to a lesser extent, kainate. The withdrawal supersensitivity was absent when 10 microM SR-95531 was coadministered with GABA during the treatment period, an event that suggests the GABAA receptors primarily produced the GABA treatment effect. This was supported further by the inability of baclofen treatment to mimic completely the treatment effect of GABA. Withdrawal from 7 days of baclofen treatment only produced a slight increase in the metabotropic effect of L-glutamate and carbachol. In addition, in untreated neurons, baclofen had no acute effect, whereas GABA inhibited the effect of L-glutamate and carbachol. The inhibitory effect of GABA was reversed by SR-95531 and was absent in neurons treated with GABA. These observations suggest the involvement of GABAA receptors and the apparent development of tolerance to GABA, respectively. Also, dependence on GABA may have occurred; the metabotropic effects of glutamate, kainate, and quisqualate were not altered in neurons maintained with GABA treatment.

GABA control of LHRH release is dependent on the steroid milieu

The effect of GABA in controlling luteinizing hormone releasing hormone (LHRH) release from isolated hypothalamic fragments containing the mediobasal hypothalamus, suprachiasmatic nucleus, and the preoptic area (MBH-SCN-POA) was tested under different hormonal conditions with an in vitro superfusion method. GABA significantly decreased LHRH release under conditions of low, chronic levels of estradiol (E 2) treatment but had no effect on tissue obtained from animals in proestrus. This suggests that the inhibitory effect of GABA is only manifested during certain stages of the estrous cycle characterized by negative feedback and low E 2.

Inhibitory effect of GABA (γ-aminobutyric acid) on the induction of long-term potentiation in guinea pig superior colliculus slices

Superior colliculus (SC) slices were prepared from guinea pigs and postsynaptic potentials (PSPs) were recorded in the superficial grey layer of the SC after electrical stimulation to the optic layer (OL). Tetanic stimulation of 50 Hz frequency and 20 s duration to the OL induced long-term potentiation (LTP) in the PSP increasing the amplitude to 140% of the original level in 30 min. Tetanic stimulation failed to induce LTP after application of γ-aminobutyric acid (GABA) (300 μM, 1 or 3 mM) to the perfusion medium. On the other hand, tetanic stimulation to the OL after application of bicuculline methiodide (1 μM), a GABA A receptor antagonist, facilitated the formation of LTP. These results indicate that GABA modulates the formation of LTP in the SC.