Infusion Of Gamma-aminobutyric Acid Research Articles

Intraventricular and Intracerebral Delivery of Anti-epileptic Drugs in the Kindling Model

A means to avoid the pharmacokinetic problems affecting the anti-epileptic drugs may be their direct intracerebroventricular (ICV) or intracerebral delivery. This approach may achieve a greater drug concentration at the epileptogenic area while minimizing it in other brain or systemic areas, and thus it could be an interesting therapeutic alternative in drug-resistant epilepsies. The objective of this article is to review a series of experiments, ranging from actute ICV injection to continuous intracerebral infusion of anti-epileptic drugs or grafting of neurotransmitter producing cells, in experimental models, especially in the kindling model of epilepsy in the rat. Acute ICV injection of phenytoin, phenobarbital or carbamacepine is able to diminish the intensity of kindling seizures, but it is also associated with a high neurologic toxicity, especially phenobarbital. Continuous ICV infusion of anti-epileptic drugs can effectively control seizures, but neurologic toxicity is not improved compared with systemic delivery. However, systemic toxicity may be improved, as in the case of valproic acid, whose continuous ICV infusion results in very low plasmatic or hepatic drug concentrations. Continuous intracerebral infusion at the epileptogenic area was studied as an alternative to minimize neurologic toxicity. Thus, intra-amygdalar infusion of gamma-aminobutyric acid (GABA) controls seizures with minimal neurotoxicity in amygdala-kindled rats. Similarly, continuous infusion of GABA into the dorsomedian nucleus of the thalamus improves seizure spread, while not affecting the local epileptogenic activity at the amygdala. Grafting of GABA releasing cells may reduce kindling parameter severity without behavioral side effects. We may conclude that ICV or intracerebral delivery of anti-epileptic drugs or neurotransmitters may be a useful technique to modulate epilepsy.

Nitrogen at raised pressure interacts with the GABA(A) receptor to produce its narcotic pharmacological effect in the rat.

Strong evidence supports the concept that conventional anesthetics, including inhalational agents and inert gases, such as xenon and nitrous oxide, interact directly with ion channel neurotransmitter receptors. However, there is no evidence that nitrogen, which only exhibits narcotic potency at increased pressure, may act by a similar mechanism. We compared the inhibitory and sedative effects of gamma-aminobutyric acid (GABA) and nitrogen pressure on locomotor activity and striatal dopamine release in freely moving rats and investigated the pharmacologic properties of the GABA-induced and nitrogen pressure-induced narcotic action using the highly selective competitive GABA(A) receptor antagonist bicuculine. Intracerebroventricular GABA infusion up to 60 micromol or exposure to nitrogen pressure up to 3 MPa decreased to a similar extent striatal dopamine release (r2= 0.899, df = 4, P < 0.01) and locomotor activity (r2 = 0.996, df = 28, P < 0.001). However, both agents only showed small effects on striatal dopamine release, reducing dopamine currents by only 12-13% at sedative concentrations. Pretreatment with bicuculline at 0.5, 1, and 2.5 pmol reduced the sedative action of GABA on locomotor activity by 10, 20, and 41%, respectively. Bicuculline in the nanomole range at 1, 2.5, and 5 nmol but not in the picomole range reduced the sedative action of nitrogen pressure by 5, 37, and 73%, respectively. Schild plot analysis is consistent with the fact that bicuculline is a competitive antagonist of both GABA and nitrogen at pressure. These results suggest (1) that the presynaptic effects of both GABA and nitrogen pressure on striatal dopamine transmission are modest and not mainly involved in their sedative action and (2) that nitrogen at increased pressure may interact directly with the GABA(A) receptor. However, because the antagonistic effect of bicuculline on nitrogen sedation only occurred at much higher bicuculline concentrations than seen with GABA, it is suggested that nitrogen does not compete for the same site as GABA.

Effects of adrenalectomy and gonadectomy on sex and stress-induced differences in bicuculline-induced convulsions.

The effects of adrenalectomy, gonadectomy and combined adrenalectomy plus gonadectomy on the previously described sex-dependent anticonvulsive effect of swim stress were studied in rats. The convulsive signs (myoclonic twitch, generalized convulsions, tonic hindlimb extension) were produced by constant i.v. infusion of gamma-aminobutyric acid(A) (GABA(A)) antagonist bicuculline, which started 15 min after termination of swim stress (10-min swim at 18-19 degrees C). Adrenalectomy decreased the threshold doses of bicuculline producing the first myoclonic twitch and the onset of generalized convulsions only in females. In adrenalectomized females, but not in males, swim stress enhanced the threshold dose of bicuculline producing generalized convulsions, but, unlike in adrenal-intact animals, it failed to enhance the dose of bicuculline producing tonic hindlimb extension. In gonadectomized stressed and unstressed animals all sex differences disappeared, and swim stress enhanced in both sexes only the threshold doses of bicuculline producing tonic hindlimb extension. Adrenalectomized plus gonadectomized animals displayed clear sex differences in doses of bicuculline necessary to produce all the convulsive signs. In the same animals swim stress postponed, especially in females, the onset of the first myoclonic twitch and generalized convulsions, but not the onset of tonic hindlimb extension. In summary, our results suggest that hormones of the adrenal and gonadal glands are only partly responsible for decreased susceptibility, especially of female rats, to the GABA(A) antagonist bicuculline. Moreover, they have demonstrated that stress produces a gender-specific anticonvulsive effect even in the animals completely deprived of steroid hormones of peripheral origin.

Effect of infusing gamma-aminobutyric acid receptor agonists and antagonists into the medial preoptic area and ventromedial hypothalamus on prolactin secretion in male sheep.

We investigated the effects of gamma-aminobutyric acid (GABA) agonists muscimol and baclofen (GABA(A) and GABA(B) agonists, respectively) and antagonists bicuculline methiodide (BMI, GABA(A) antagonist) or 2-hydroxysaclofen (SAC) and CGP 55845A (GABA(B) antagonists) on prolactin (PRL) secretion in castrated rams. The drugs were applied by microdialysis into either the medial preoptic area (mPOA) or ventromedial hypothalamus (VMH). Dialysis of baclofen into the mPOA significantly increased mean PRL (p < 0.05), whereas SAC caused a small, but significant decrease (p < 0.01). Dialysis of either muscimol or BMI into the mPOA had no effect on prolactin. In the VMH, baclofen significantly increased (p < 0.01) mean PRL but SAC and CGP 55845A were ineffective, whereas dialysis of either muscimol or BMI increased mean prolactin (p < 0.01). These results show that infusion into the mPOA of drugs that affect GABA(B) receptor alter PRL release, whereas infusion of a GABA(A) agonists and antagonist was without effect on PRL release. In contrast, infusion of both GABA(A) and GABA(B) agonists and a GABA(A) antagonist into the VMH altered PRL secretion. This suggest that GABAergic neurons in both regions participate in regulating PRL secretion, but by different receptor systems.

The effect of infusion of putrescine and gamma amino butyric acid on the intake of steers offered grass silage containing three levels of lactic acid

An experiment was carried out to assess the role of putrescine (P) and gamma amino butyric acid (GABA) on the intake of grass silage containing three levels of lactic acid. The experimental design involved a nine period, 3 × 3 factorial changeover design experiment, with each period of 14 days duration. Putrescine and GABA were applied at a rate of 6 g kg −1 dry matter (DM), dissolved in 4 I water and intraruminally infused using a peristaltic pump with the amount infused varied throughout the day by means of an electronic time-switch attached to the infusion pump. (This was an attempt to match the amount infused with the amount the animal would have consumed if the treatment had been added to the diet.) The basal diet consisted of a well fermented grass silage with 116 g lactic acid kg −1 DM. Lactic acid was added to the basal diet at levels of 0, 50 and 100 g kg −1 DM to give final lactic acid concentrations of 116 (L1), 160 (L2) and 188 (L3) g kg −1 DM, respectively. No significant interactions between infusion treatment and level of lactic acid were observed on intake, eating behaviour, rumen fermentation parameters or dry matter, nitrogen, modified acid detergent fibre (MADF) or acid detergent fibre (ADF) degradability characteristics of silage. Main treatment effects only are presented. Putrescine and GABA did not significantly ( P > 0.05) affect dry matter intake or feeding behaviour. L2 significantly increased total time spent eating per day ( P < 0.05), reduced daily eating rate ( P < 0.001) and reduced mean eating rate during the secondary meals ( P < 0.001). Infusion of P increased rumen fluid pH ( P < 0.05). GABA infusion reduced the molar proportion of propionic acid ( P < 0.05) and increased total i-butyric acid concentration ( P < 0.05). L3 increased total concentration of n-butyric acid ( P < 0.05); L2 and L3 increased the molar proportion of n-butyric acid ( P < 0.05). Lactic acid treatment had no significant effect ( P > 0.05) on DM, N or MADF degradability characteristics. However, L3 reduced NDF degradability ( P < 0.05). Infusion treatment had no significant effect ( P > 0.05) on DM, MADF or NDF degradability characteristics. However, P reduced nitrogen degradability ( P < 0.01).

The effect of intraruminal infusions of amines and gamma amino butyric acid on rumen fermentation parameters and food intake of steers offered grass silage

An experiment was carried out to assess the role of amines and gamma amino butyric acid (GABA) in food intake control with steers offered grass silage based diets. The effect of the amines, putrescine (P) and tyramine (T), and GABA (G) on voluntary food intake were investigated at three levels of addition: 2 g kg −1 DM (1); 4 g kg −1 DM (2) and 6 g kg −1 DM (3). An additional ‘control’ treatment involved infusion of water. A ten period changeover design experiment was used, with four steers, mean liveweight 597 ± 38 kg, fitted with rumen cannulae. The basal diet consisted of an extensively fermented grass silage offered ad libitum. Each period was of one week duration. Following a 4 day preliminary period, amines, GABA or water were infused into the rumen for 3 days for 8 h per day. Daily dry matter intake was recorded in addition to intakes at various time intervals throughout the day. Rumen samples were also taken throughout a 24 h period on Day 3 of each experimental period. No significant ( P > 0.05) treatment effects on DM intake were observed with dry matter intakes (g kg −1 LW) as follows: 13.1 (C); 13.7 (P1); 13.2 (P2); 12.7 (P3); 13.7 (G1); 13.8 (G2); 13.9 (G3); 14.2 (T1); 14.1 (T2) and 13.2 (T3) SED (0.68). Tyramine infusion at 6 g kg −1 DM significantly ( P < 0.05) increased rumen fluid pH, GABA infusion at 2 g kg −1 DM significantly ( P < 0.05) reduced the proportion of propionate and tyramine infusion at 2 and 6 g kg −1 DM significantly ( P < 0.05) increased isovalerate proportion in rumen fluid. These results suggest that infusion of putrescine, tyramine and GABA into the rumen at levels up to 6 g kg −1 DM intake, do not significantly affect voluntary food intake with steers offered an extensively fermented grass silage.

The effects of either dietary additions or intraruminal infusion of amines and juice extracted from grass silage on the voluntary intake of steers offered grass silage

An experiment was carried out to assess the possible role of amines and other end products of silage fermentation on food intake of steers. In total, 6 treatments were examined in a 9 period changeover design experiment with 4 steers, mean live weight 520 kg, fitted with rumen cannula. Each period was one week long. The 6 treatments comprised putrescine (P); cadaverine (C); gamma amino butyric acid (GABA); putrescine + cadaverine + gamma amino butyric acid (P + C + GABA); expressed juice from grass silage (S); water (w) control. These were either added to the diet (D) or infused directly into the rumen (I). Amines were applied at a rate of 2 g kg −1 DM and were dissolved in water for infusion and addition to the diet. The basal diet consisted of a restricted fermentation grass silage, from which juice was expressed prior to feeding using a hydraulic press. Within each 7 day period, animals were offered a standard silage for the first 4 days following which treatments were applied for 3 days, with total dry matter intake recorded each day of the 3 day treatment period. On day 2, intakes were measured at 5 min intervals for the first 3 h following feeding and every 2 h for the next 10 h. Dry matter intakes (g kg −1 LW) for each of the treatments were as follows: 10.2 (W,I); 10.9 (W,D); 11.6 (S,I); 12.1 (S,D); 9.0 (GABA,I); 10.5 (GABA,D); 10.8 (P,I); 10.4 (P,D); 12,2 (C,I); 10.4 (C.D); 11.0 ([P + C + GABA],I); 10.3 ([P + C + GABA],D); SEM = 0.780. These results indicate that intraruminal infusion or dietary addition of amines, GABA or silage juice at levels up to 2 g kg −1 DM, had no significant ( P > 0.05) effect on the voluntary food intake of steers offered grass silage.

Role of amino acids in salivation and the localization of their receptors in the rat salivary gland.

The distribution of gamma-aminobutyric acid (GABA) receptor subunits such as GABAAR-gamma 1 and GABAAR-gamma 2, and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) type receptor subunits such as GluR-1, GluR-2/3 and GluR-4, and N-methyl-D-aspartic acid (NMDA) type subunits such as NR1 were investigated by immunocytochemistry. Furthermore, the roles of these amino acids, GABA and glutamate, on salivation were analyzed in the rat submandibular and sublingual glands. Some similarities were observed in the distribution patterns of GABAA type receptors and AMPA receptors. In the submandibular ganglion cells, collecting ducts and striated ducts, these subunits were expressed strongly; however, there were some differences in their expression patterns between the submandibular and sublingual gland acinar cells. Since these receptor subunits were expressed in the acinar cell bodies of the submandibular gland, they were not expressed in the acinar cells but were expressed in the myoepithelial cells in the sublingual gland. On the other hand, no NR1 expression was observed. To examine the roles of GABA and glutamate in salivation, the submandibular and sublingual glands were perfused partially with Ringer's solution via a facial artery to avoid systemic influence, and substrates were infused into the perfusion solution. No salivary secretion was evoked by GABA or glutamate infusion in the absence of electrical stimulation (2-3 V, 5 ms, 20 Hz). Salivary flow evoked by electrical stimulation of the chorda-lingual nerve caused significant inhibition by GABA (10(-6), 10(-5), 10(-4) and 10(-3) M) and the GABAAR agonist muscimol 10(-3) and 10(-6) M) (n = 6, P < 0.05). Such GABA-induced inhibition was antagonized by the GABAAR antagonists bicuculline (BCC; 10(-6) and 10(-3) M) and picrotoxin (PTX; 10(-6) and 10(-3) M). On the other hand, salivary flow evoked by electrical stimulation (8-10 V, 5 ms, 20 Hz) of the superior cervical ganglion (SCG) was not affected by GABA. While high doses of glutamate (10(-1) M) and NMDA (10(-1) M) showed no effects on salivary flow despite application of electrical stimulation, AMPA at a high concentration (10(-1) M) significantly inhibited salivary secretion (n = 6, P < 0.05). These studies revealed that inhibitory and excitatory amino acid receptors such as GABAA and AMPA type receptors are coexpressed in the rat salivary glands, and that GABA inhibits salivary secretion via GABAA receptors which may act with acetylcholine. However, the role of glutamate in salivation remains unclear despite the presence of AMPA type receptors. The present findings suggest that glutamate does not act alone but with other substances such as peptides and/or other amino acids.

Decrease of glutamate decarboxylase activity after in vivo cortical infusion of γ-aminobutyric acid

gamma-Aminobutyric acid (GABA) levels and the activity of glutamate decarboxylase were measured in homogenates of rat brain cortical tissue, at different times after chronic intracortical infusion of GABA in vivo during 2, 6 or 24 h. Cortical electrical activity was also recorded. As previously described, about 1 h after cessation of the infusion epileptic discharges were observed (GABA-withdrawal syndrome), which lasted for several days. At zero time after cessation of the infusion, before the appearance of seizures, GABA levels were increased 3-6-fold and glutamate decarboxylase activity was decreased 27-48% in the infused cortex, as compared to the contralateral cortex or to tissue from control intact rats. During epileptic discharges GABA levels gradually returned to normal values. In contrast, glutamate decarboxylase activity remained decreased during seizures and returned to normal only after recovery from the GABA-withdrawal syndrome. These results suggest that the persistent decrease in the activity of the decarboxylase is due probably to a lowered amount of the enzymatic protein, occurring as a consequence of a temporarily elevated intracellular GABA concentration. The decreased rate of GABA synthesis might be involved in the pathophysiology of the GABA-withdrawal syndrome.

Noradrenaline mediates paradoxical effects on rat neocortical neurons after GABA withdrawal

1. The aim of the present study was to determine the role of noradrenergic neurotransmission in neuronal activities intracellularly recorded in neocortical slices obtained from rats presenting the gamma-aminobutyric acid (GABA) withdrawal syndrome (GWS), a focal epilepsy consecutive to the interruption of a chronic intracortical GABA infusion into the somatomotor cortex. Neurons recorded in the epileptic focus area (n = 52) were bursting or nonbursting cells. Intrinsic bursting (IB, n = 20) cells presented bursts of action potentials (APs) to an intracellular depolarizing current injection and paroxysmal depolarization shifts (PDSs) to white matter stimulation. Synaptic bursting (SB, n = 22) cells presented only PDSs. Nonbursting (NB, n = 10) cells presented no burst after either synaptic stimulation or depolarizing current injection. Results were compared with those obtained from NB neurons (n = 4) recorded in slices from saline-infused rats. 2. In all of the recorded neurons, bath application of norepinephrine (NE, 10 and 100 microM) provoked a depolarization (1-5 mV) associated with a decrease in input K+ conductance having a mean reversal potential at -90 to -102 mV, not significantly different for bursting and nonbursting cells. This reversal potential differed from that of Cl(-)-mediated inhibitory postsynaptic potentials (-70 mV) elicited in NB cells by electrical stimulation of the white matter. 3. In IB cells, the NE-induced depolarization replaced the intrinsic bursts by a sustained repetitive discharge of single APs and caused intrinsic bursts to appear during previously subthreshold depolarizing current pulses. These NE-increased activities were abolished by dihydropyridine nitrendipine (1 microM) and by Cd2+ (0.5 mM) or Co2+ (2 mM), thus confirming that Ca2+ currents contribute to burst generation in IB cells. 4. In both NB and SB cells recorded in slices from GWS rats, NE provoked the appearance of intrinsic bursts of APs during steps of depolarizing current injections. In addition, in NB cells, NE caused synaptic bursts to appear after white matter stimulation. These NE-induced bursts were dihydropyridine (nitrendipine, 1 microM)- and Cd2+ (0.5 mM)- or Co2+ (2 mM)-sensitive and were related to an increased AP-afterdepolarization. The fast AP-afterhyperpolarization was not affected by NE. In NB cells recorded in slices from saline-infused rats (n = 4) NE did not provoke the appearance of bursts even when stimulation intensity was increased up to three times.(ABSTRACT TRUNCATED AT 400 WORDS)

Laterality of the gonadotrophin releasing hormone pulse generator in the rat: interaction of neurons located in both preoptic areas as a basic feature.

Though the pivotal importance of pulsatile gonadotrophin releasing hormone (GnRH) release from the hypothalamus for sufficient luteinizing hormone (LH) secretion is well recognized, the mechanisms causing GnRH pulses are still largely unknown. In the rat, the GnRH neurons are concentrated in the preoptic area, which is a bilateral structure of the anterior hypothalamus. This distribution into two distinct brain areas raises the question of whether the GnRH neurons in both preoptic areas are simultaneously or alternatively active and whether an interaction between both areas is required to generate GnRH pulses. We addressed these questions by unilateral manipulations of the activity of GnRH neurons using either electrical lesion or local infusion of gamma-aminobutyric acid (GABA), a neurotransmitter known to inhibit LH release via a hypothalamic action. Acute lesion of one preoptic area caused complete cessation of pulsatile LH release which was restored after 48 h. An acute inhibition of LH release was also observed after unilateral preoptic application of GABA. It is concluded that an interaction of both preoptic areas is mandatory for GnRH pulse generation. However, we suggest that the pacemaker function is not intrinsic to GnRH nor GABA neurons but the cross-talk between these neurons within and between both preoptic areas results in episodic GnRH release.

Local cerebral glucose utilization during chronic infusion of GABA into the nucleus basalis magnocellularis of rats

The metabolic effects of gamma aminobutyric acid (GABA) unilaterally infused for 24 h into the nucleus basalis magnocellularis (NBM) of rats were studied using the 2-deoxyglucose method for measuring local cerebral metabolic rates for glucose (CMRglc). These results demonstrated that GABA infusion at the concentration of 100 μg·μl −1 induced a strong ipsilateral depression in glucose utilization in the NBM and in some surrounding structures (e.g., the lateral hypothalamic area, thalamus, and amygdala) as well as in the major cortical targets of NBM neurons (i.e., the frontal, parietal, and temporal cortices). On an other hand, GABA infusion at the concentration of 10 μg·μl −1 had no significant effect on cerebral metabolism compared to the saline-infused rats. Nevertheless, both GABA at the concentration of 10 μg·μl −1 and saline induced a reduction of cerebral metabolism in the infused NBM, compared to the contralateral noninfused NBM. It was also demonstrated that saline pretreatment reduced the metabolic depression induced by GABA infusion at the highest concentration. When GABA was infused at the concentration of 100 μg·μl −1, a slight reduction of CMRglc was observed in the hemisphere contralateral to the infused one, mainly in NBM target areas. In view of our previous behavioral data, these results suggest among others, that there is no necessary relationship between cerebral glucose metabolism and behavioral expression.

Burst generation in neocortical neurons after GABA withdrawal in the rat

1. gamma-Aminobutyric acid (GABA) withdrawal syndrome (GWS) represents a particular model of focal epilepsy consecutive to the interruption of a chronic intracortical GABA infusion and is characterized by the appearance of focal epileptic electroencephalographic (EEG) discharges and localized clinical signs on withdrawal of GABA. Effects of Ca2+ channel blockers and N-methyl-D-aspartate (NMDA) antagonists were evaluated in living rats presenting a GWS after interruption of a 5-day GABA infusion into the somatomotor cortex and in neocortical slices obtained from such rats. Bursting properties and morphology of neurons were also analyzed in slices. 2. In living rats, the noncompetitive NMDA antagonist phencyclidine [1-(1-phenylcyclohexyl)piperidine] and the Ca2+ antagonist flunarizine [E-1 (bis(4fluorophenyl)methyl)-4(3phenyl2-propenyl)-piperazine] were administered systemically to two groups of rats. Rats in the first group (n = 12) were injected with the drug 30-60 min before discontinuation of the GABA infusion. In this case, phencyclidine (10 mg/kg ip) prevented the development of GWS (n = 5), whereas flunarizine (40 mg/kg ip) had no consistent effect on the GWS appearance and characteristics (n = 7). Rats in the second group (n = 12) were injected 60-90 min after GABA discontinuation, i.e., during a fully developed GWS. In that case, neither drug suppressed GWS. 3. Neuronal activities in the epileptic focus were studied in slices with conventional intracellular recording and stimulation techniques. From the 65 neurons recorded, 29 responded with EPSPs and paroxysmal depolarization shifts (PDSs) to white matter stimulation (synaptic bursting or SB cells). Nineteen other neurons presented, in addition to synaptically induced PDSs, bursts of action potentials (APs) induced by intracellular depolarizing current injection (intrinsic bursting or IB cells). The remaining 17 neurons presented no bursting properties to either synaptic stimulation or depolarizing current injection (nonbursting or NB cells). 4. The recorded neurons were located 0.7-1.2 mm distant from the lesion because of the penetration of the GABA infusion cannula. Intracellular injection of neurons (n = 4) with biocytin or Lucifer yellow revealed that both SB and IB neurons were large, spiny pyramidal neurons localized in layer V of the sensorimotor cortex. 5. Bath application of the selective antagonist of NMDA receptors DL-2amino-5phosphonovalerate or DL-2amino-7phosphonoheptanoate (10-50 microM) reversibly reduced the amplitude (by 25-50%) and the duration (by 20-25%) of PDSs in all cases (n = 17).(ABSTRACT TRUNCATED AT 400 WORDS)

Avian locomotion activated by brainstem infusion of neurotransmitter agonists and antagonists. II. gamma-Aminobutyric acid.

The companion article (Sholomenko et al. 1991) described the brainstem locomoter regions in the bird where direct intracerebral injection of a number of putative excitatory neurochemicals, including cholinergic agonists, N-methyl-d-aspartate (NMDA) and Substance P, evoke locomotion. Using the same experimental protocol, this study focuses on the locomotor effects following discrete brainstem injections of the inhibitory neurotransmitter, gamma-aminobutyric acid (GABA), and some of GABA agonists and antagonists. Brainstem regions that were electrically and chemically stimulated included the ventromedial medullary reticular formation, the pontobulbar locomotor strip of the dorsolateral pons and medulla, the pontine reticular formation, and the mesencephalic reticular formation. Locomotion was evoked after the injection of the GABA antagonists picrotoxin (a GABAA receptor antagonist) and bicuculline (GABAA antagonist) into several brainstem locomoter regions. Brainstem stimulated locomotion (both chemically and electrically induced) could be transiently blocked by intracerebral infusion of GABA and irreversibly blocked by muscimol (GABAA agonist). Our avian results are similar to those described for mammals and provide support for the suggestion that motor circuitry, at least at brainstem levels, is similar in all vertebrates.

Isolation of Gamma-Amino Butyric Acid from Pig Hypothalami and Demonstration of its Prolactin Release-Inhibiting (PIF) Activityin VivoandinVitro*

A non-retarded fraction with prolactin-release inhibiting factor (PIF) activity obtained by chromatography of a concentrate of porcine hypothalami on carboxymethyl-cellulose was chromatographically distinct from catecholamines. This fraction was purified further by six steps involving chromatography on Sephadex G-25, countercurrent distribution, free-flow electrophoresis, and chromatography on triethylaminoethyl cellulose. The PIF-active substance was isolated and identified as gamma-amino-butyric acid (GABA) by: 1) amino acid analyses using sodium as well as lithium-based buffers for resolution of biological fluids, 2) thin-layer chromatography of underivatized material as well as phenylthiocarbamyl derivatives, and 3) mass spectroscopy. Natural and synthetic GABA inhibited prolactin, but not LH release in vitro from isolated rat pituitary halves at doses as low as 0.1 microgram/ml. The inhibition was proportional to the dose; natural and synthetic GABA possessed identical PIF activity. Synthetic GABA also decreased prolactin release in monolayer cultures of rat pituitary cells and inhibited TRH-stimulated prolactin release. The inhibition of prolactin release in vitro by GABA could not be blocked by perphenazine, which inhibits PIF activity of catecholamines. GABA also suppressed prolactin release in vivo, although large doses were needed. Either rapid iv injection or infusion of GABA in doses of 1 to 100 mg in rats significantly decreased serum prolactin levels, which were previously elevated by pretreatment with monoiodotyrosine perphenazine, chlorpromazine, haloperidol, or sulpiride. beta-hydroxy GABA significantly depressed prolactin release, but beta-(p-chlorophenyl)-GABA (Lioresal, CIBA) and 4 other analogs of GABA were not effective in vivo and/or in vitro. The results indicate that GABA can inhibit prolactin release by a direct action on the pituitary gland, but whether this effect is physiologically meaningful still remains to be determined.

Effects of parenterally administered gamma-aminobutyric acid on the general behavior of the young chick

RECENTLY we reported that intravenous infusion of gamma-amino butyric acid (GABA) to young chicks (7–14 days post hatch) markedly altered the electroretinogram (ERG) and suppressed the bioelectrical potential in the optic tectum evoked by photic stimulation (1). These results, together with a briefly noted behavioural observation (2) that intraperitoneal injection of GABA to baby chicks, at dose levels sufficient to elevate its normal concentration in the brain, produced overt signs of uncoordinated motor activity suggests that parenterally administered GABA has a marked central action on young chicks. However, in a similar preparation, Kramer et al. (3) encountered no striking effects resulting from administration of GABA. In view of this seeming contradiction we made a more detailed study of the behavioural responses of young chicks to the parenteral administration of GABA.