Injection Of Bicuculline Methiodide Research Articles

Mechanism of the cardiovascular effects of the GABAA receptors of the ventral tegmental area of the rat brain

The ventral tegmental area (VTA) contains GABA terminals involved in the regulation of the cardiovascular system. Previously, we demonstrated that blocking GABAA but not GABAB receptors produced a pressor response accompanied by marked bradycardia. This study was performed to find the possible mechanisms involved in these responses by blocking ganglionic nicotinic receptors, peripheral muscarinic receptors or peripheral V1 vasopressin receptors.Experiments were performed on urethane anesthetized male Wistar rats. Drugs were microinjected unilaterally into the VTA (100nl). The average changes in mean arterial pressure (MAP) and heart rate (HR) were compared between pre- and post-treatment using paired t-test.Injection of bicuculline methiodide (BMI), a GABAA antagonist, into the VTA caused a significant increase in MAP and a decrease in HR. Administration (i.v.) of the nicotinic receptor blocker, hexamethonium, enhanced the pressor response but abolished the bradycardic response to BMI, which ruled out involvement of the sympathetic nervous system. Blockade of the peripheral muscarinic receptors by homatropine (i.v.) abolished the bradycardic effect of BMI, but had no effect on the pressor response, indicating that bradycardia was produced by the parasympathetic outflow to the heart. Both the pressor and bradycardic responses to BMI were blocked by V1 receptor antagonist (i.v.), indicating that administration of BMI in the VTA disinhibited the release of vasopressin into the circulation.In conclusion, we demonstrated that GABAergic mechanism of the VTA exerts a tonic inhibition on vasopressin release through activation of GABAA receptors. The sympathetic system is not involved in the decrease of blood pressure by GABA of the VTA.

Vasopressin and sympathetic systems mediate the cardiovascular effects of the GABAergic system in the bed nucleus of the stria terminalis

The bed nucleus of the stria terminalis (BST) is an important part of the limbic system. It has been shown that chemical stimulation of the BST elicited cardiovascular depressive and bradycardic responses. It was also demonstrated that GABA is present in the BST, though its role in cardiovascular control is not yet understood. This study was performed to find the effects of GABA receptor subtypes in the BST on cardiovascular responses and to find the possible mechanisms that mediate these responses in urethane-anesthetized rats. Microinjection of muscimol (500 pmol/100 nl), a GABA A agonist, into the BST produced a weak unsignificant decrease in the mean arterial pressure (MAP) and heart rate (HR). Injection of bicuculline methiodide (BMI, 100 pmol/100 nl), a GABA A antagonist, caused a significant increase in the MAP (41.3 ± 5.1 mmHg) as well as in the HR (33.2 ± 5.6 beats/min). Injection of two doses (500 and 1000 pmol/100 nl) of phaclofen, a GABA B antagonist, produced no significant change in either MAP or HR. Administration (i.v.) of the muscarinic receptor blocker, homatropine methyl bromide had no effect on the magnitude of mean arterial pressure or heart rate responses to BMI. This suggests that the parasympathetic system is not involved in these responses. However, administration (i.v.) of the nicotinic receptor blocker, hexamethonium bromide had no effect on the magnitude of mean arterial pressure response but abolished heart rate response to BMI. This suggests that the sympathetic system is involved in the bradycardic effect of GABA. On the other hand, administration (i.v.) of a selective vasopressin V 1 receptor antagonist abolished the pressor effect of BMI, which indicates that the GABAergic system of the BST decreases the arterial pressure via tonic inhibition of vasopressin release. In summary, we demonstrated, for the first time, that GABA exerts its influence in the BST through the activation of GABA A, but not GABA B, receptors that, in turn, tonically inhibit vasopressin release and sympathetic outflow to the heart.

Inhibition of central GABA A receptors enhances hepatic glucose production and peripheral glucose uptake

Previous studies have demonstrated that intracerebroventricular (ICV) injection of bicuculline methiodide (BMI), a gamma-amkrobutyric acid receptor antagonist, increases plasma glucose concentrations. The purpose of the present study was to determine whether the hyperglycemic response was due to an increased rate of hepatic glucose production (HGP) or a change in the rate of glucose utilization. In vivo glucose flux was assessed in catheterized, conscious overnight fasted rats using [3- 3H]glucose. ICV injection of BMI (10 nmol) increased glucose levels 60% after 30 min. This hyperglycemia resulted from a rapid increase in HGP that exceeded an increased rate of glucose utilization. No alteration in the glucose metabolic clearance rate, an index of the avidity of the body's tissues for glucose, was detected in BMI-Injected rats. BMI enhanced both hepatic gluconeogermals and glycogenolysis, since the reduction in War glycogen (19 μmol/g liver) could not totally account for all of the increased HGP. These metabolic alterations were associated with sustained increases in circulating concentrations of corticosterone, giucagon and catecholamines. Prior adrenalectomy completely abolished the BMI-induced increase in glucose flux and the reduction in tissue glycogen, despite the persistent hyperglucagonemia. These data indicate that, in the fasted condition, the hyperglycemia produced by central administration of BMI results from an increased rate of HGP (both gluconeogenesis and glycogenolysis) and not a reduction in the ability of tissues to use glucose. The concomitant elevation in glucose disposal was the result of an increased mass action effect. The enhanced glucose metabolic response to BMI appears mediated exclusively by an increased secretion of epinephrine from the adrenal medulla.

GABA receptors in the region of the dorsomedial hypothalamus of rats regulate anxiety in the elevated plus-maze test. II. Physiological measures

In the previous report, we had shown that blockade and enhancement of GABA A receptors in the DMH of rats increased or decreased the level of anxiety, respectively, as measured by the elevated plus-maze test. The present study was conducted to assess the effects of enhancing GABA A neurotransmission in the DMH of rats on the physiological concomitants of anxiety such as increased in heart rate (HR), blood pressure (BP) and plasma norepinephrine (NE) levels while the animals were placed on the elevated plus-maze. Male Sprague-Dawley rats were equipped with arterial and venous catheters and stereotaxically implanted with microinjection cannulae in the cardiostimulatory region of the DMH where injection of bicuculline methiodide (BMI) elicited increases in heart rate under anesthesia. After recovery, rats were injected with either saline or the GABA A agonist muscimol and their HR, BP and plasma NE responses were measured when confined in the open or the closed arm of the elevated plus-maze. Injection of muscimol into the DMH reduced the increases seen in HR, BP and plasma NE when the rats were confined to either the closed or the open arms in addition to decreasing ‘anxiety’ in the plus-maze. Injection of muscimol into the areas of the hypothalamus surrounding the DMH did not significantly affect the changes in HR, BP and plasma NE in the plus-maze. Blocking the changes in HR and BP elicited by microinjecting GABAergic drugs into the DMH of rats, with systemic injections of a combination of atropine and the beta-blocker atenolol, did not block the behavioral effects of the GABAergic drugs in the plus-maze test.

Zona incerta-lateral hypothalamus as an output structure for impulses involved in neuroleptic drug-induced catalepsy.

Our previous studies showed that the neuronal impulses connected with catalepsy, which have their origin at dopamine D2 receptors in the ventro-rostral part of the nucleus caudatus-putamen in rats, are conveyed to the zona incerta-lateral hypothalamic region. The aim of the present study was to investigate the route of the neuronal impulses between these structures. The experiments were carried out on rats with cannulae chronically implanted in the brain structures. We showed that (1) bilateral injection of bicuculline methiodide (5-50 ng) into the ventro-medial part of the globus pallidus (GPv) and (2) bilateral injection of muscimol (2.5-25 ng) into the substantia nigra pars reticulata (SNR) inhibit, in a dose dependent manner, the catalepsy induced by sulpiride (1 microgram) administered bilaterally into the ventro-rostral part of the nucleus caudatus-putamen. It was also demonstrated that muscimol (25 ng), injected bilaterally into the ventro-medial part of the globus pallidus, induces catalepsy which, in turn, is dose-dependently inhibited by either (1) muscimol (5-25 ng) injected into the substantia nigra pars reticulata, or (2) bicuculline (1.0-2.5 ng) injected into the zona incerta-lateral hypothalamus (ZI-LH). Moreover, even a dose as high as 50 ng of bicuculline, injected into the ventro-medial part of the globus pallidus, had no significant effect on the locomotor activity of rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Amygdaloid kindling with bicuculline methiodide in rats

The effect of repeated intraamygdaloid injection of bicuculline methiodide (BM) was studied in rats. Chemitrodes for both microinjection and electrographic recording were implanted into the left basolateral amygdala. Two weeks after the surgery, a subconvulsive dose of BM (0.2 or 0.4 nmol) was administered through the chemitrodes every fourth day. Repeated injections caused a progressive seizure development which was comparable to that seen with electrical kindling. The kindling effect persisted after a 6-month interruption of the stimulation. When a mixture of BM and GABA agonist (GABA 20 nmol, muscimol 2 nmol, or baclofen 5 nmol) was injected into the amygdala of kindled rats, seizure activity was markedly suppressed to stage 0 or 1. On the other hand, an intraamygdaloid injection of picrotoxin (0.8 nmol) brought about the same seizure as induced by BM, whereas no seizure was observed with strychnine (4 nmol). No histological change specific to this kindling was detected. The present results indicate that chemical kindling can be induced by repeated local injection of BM into the amygdala, and that the mechanism underlying this kindling is closely associated with local postsynaptic GABA receptors in the amygdala. This GABAergic system may also be important in other types of kindling.

Origin of tonic GABAergic inputs to vasopressor neurons in the subretrofacial nucleus of the rabbit

The aims of this study were to determine (1) whether the vasomotor effects reflexly elicited by baroreceptor stimulation are dependent upon γ-aminobutyric acid (GABA) receptors in the subretrofacial (SRF) nucleus in the rostral ventrolateral medulla; (2) the extent to which inputs other than those arising from peripheral baroreceptors, or transmitted via the nucleus tractus solitarius (NTS), contribute to the tonic GABAergic inhibition of SRF vasopressor cells. Following bilateral injection of a mixture of the GABA antagonist bicuculline methiodide (500 pmol) and GABA agonist muscimol (500 pmol) into the SRF nucleus, the sympathoinhibitory response normally evoked by a rise in arterial pressure (induced by inflating an aortic cuff) was abolished in 4 out of 8 rabbits and reduced in the remainder. For the whole group, the mean reduction in this response was 71%. In other experiments, the pressor response produced by injection of bicuculline methiodide into the SRF nucleus was still present after (1) destruction of the intermediate portion of the NTS, and (2) complete removal of the brain rostral to the pons. We conclude that (1) an inhibitory GABAergic input into the SRF nucleus is an important component of the central pathways mediating baroreceptor inhibition of sympathetic vasomotor tone; (2) the SRF nucleus also receives tonic GABAergic inputs that are intrinsic to the lower brainstem and are independent of baroreceptor or other cardiovascular inputs relayed by the NTS.

GABA stimulation and blockade in the hypothalamus and midbrain: Effects on feeding and locomotor activity

Microinjections of the gamma-aminobutyric acid (GABA) antagonist, bicuculline methiodide (BM) (100 ng), into the anterolateral hypothalamus (LH) increased ingestion of sweet milk. A subsequent injection of BM 48 hrs. later produced a type of kindling effect consisting of feeding related automatisms, such as gnawing and bitting. The behavioral effects of injections of 100 ng of GABA into the LH were variable. GABA injections into the ventromedial hypothalamus (VMH) reliably increased food intake. GABA injections into the origin of the nigrostriatal dopamine (DA) neurons in the substantia nigra (SN) suppressed it. Similar injections into the origin of the mesolimbic DA cells in the ventral tegmental area (VTA) had no effect on feeding behavior. Following BM injections into the SN, a moderate increase in tilt box activity was observed. A second injection of the GABA blocker 6 days later exaggerated this effect. Short latency extreme hyperactivation was accompanied by unidirectional barrel rolling which persisted until blocked by local injections of GABA.