GABAergic modulation of neurons in the nucleus of the solitary tract with ascending projections to the subfornical organ in the rat

Twenty-five neurons in the region of the nucleus of the solitary tract (NTS) were antidromically activated by electrical stimulation of the subfornical organ (SFO) in male rats under urethane anesthesia. Microiontophoretically applied bicuculline, a gamma-aminobutyric acid (GABA)(A) antagonist, but not phaclofen, a GABA(B) antagonist, attenuated the post-antidromic inhibitory response evoked by SFO stimulation of approximately two-third (n=17) of identified neurons, indicating the existence of recurrent inhibitory systems through GABA(A) receptors. Iontophoretically applied GABA decreased the spontaneous activity of all identified neurons, and the GABA-induced inhibition was prevented by simultaneously applied bicuculline, but not by phaclofen. Activation of peripheral baroreceptors, achieved by rising arterial blood pressure with an intravenous infusions of phenylepherine, suppressed the activity of the majority (n=20) of identified neurons. The inhibitory response of identified neurons (n=7) to baroreceptor activation was partially antagonized by iontophoretically applied bicuculline, but not by phaclofen. These results imply that GABAergic mechanisms may modulate the baroreceptor reflex acting on GABA(A) receptors of NTS neurons with ascending projections to the SFO in the region of the NTS.     

Inhibitory inputs to the subfornical organ from the AV3V: involvement of GABA.

Action potentials were recorded extracellularly from single neurons in the subfornical organ (SFO) of the pentobarbital-anesthetized cat following stimulation of the regions surrounding the anteroventral third ventricle (AV3V). Of 328 SFO neurons studied, 103 were antidromically activated, showing direct projections from the SFO to the AV3V. However, the major effects of stimulations of the AV3V on SFO neurons were orthodromic inhibition; almost 30% of SFO neurons were inhibited by various sites in the AV3V, while a smaller proportion of cells were excited. Local application of bicuculline, an antagonist for GABA, attenuated the inhibitory responses induced by stimulation of the AV3V in seven out of eight neurons tested. Application of GABA inhibited 16 out of 24 neurons, while that of bicuculline alone excited 11 out of 26 neurons, suggesting the tonic inhibitory action of GABA on some SFO neurons. On the other hand, application of kynurenic acid, a nonspecific antagonist for the excitatory amino acids, did not affect the excitatory responses induced by stimulation of the AV3V, but kynurenic acid itself inhibited 6 out of 18 neurons tested. Application of glutamate excited most SFO neurons. This suggests that the excitatory amino acids may be the transmitter(s) of interneurons in the SFO but may not mediate the excitation from the AV3V.     

GABAergic systems in the nucleus tractus solitarius regulate noradrenaline release in the subfornical organ area in the rat

Previous studies have shown that catecholaminergic neurons in the nucleus tractus solitarius (NTS) with ascending projections to the subfornical organ (SFO) are highly sensitive to gamma-aminobutyric acid (GABA). To clarify the role of the GABAergic system in the NTS in the regulation of the activity of noradrenergic NTS projections to the SFO, the present study was carried out to investigate the effects of local administration (50 nl) of GABA, the GABA(A) agonist muscimol, the GABA(B) agonist baclofen, the GABA(A) antagonist bicuculline or the GABA(B) antagonist phaclofen into the NTS on the release of noradrenaline (NA) in the region of the SFO using microdialysis techniques in rats under urethane anesthesia. Microinjections of GABA (10(-4) - 10(-2) M) into the region of the NTS significantly decreased the NA release in the SFO area. Injections of either muscimol (10(-4) - 10(-2) M) or baclofen (10(-5) - 10(-3) M) into the NTS region significantly attenuated the NA release in the SFO area. Injections of bicuculline (10(-5) and 10(-4) M), but not phaclophen (10(-6) - 10(-4) M), into the NTS region significantly enhanced the NA release in the SFO area, suggesting that the GABAergic system in the NTS may tonically inhibit the NA release in the SFO area through a GABA(A) receptor mechanism. Neither injection of these drugs in any of the doses used in this study into the NTS region caused any significant changes in the NA release in the sites away from the SFO. Injections of vehicle (50 nl) into the NTS region had no significant effect on the NA release in either the SFO area or the sites away from the SFO. These results suggest that the GABAergic system in the NTS may serve to decrease the release of NA in the SFO area and the two types of GABA receptors are involved in the modulation of the NA release.     

Activation of serotonergic pathways from the midbrain raphe system to the subfornical organ by hemorrhage in the rat

The role of serotonergic neural pathways from the midbrain raphe nuclei to the subfornical organ (SFO) in the central regulation of cardiovascular function and body fluid balance was investigated in adult male rats under urethane anesthesia. Eleven neurons in the dorsal raphe nucleus (DR) were antidromically activated by electrical stimulation of the SFO. Of these neurons, 6 displayed an excitatory response following hemorrhage (10 ml/kg bwt) while the remaining 5 neurons were unresponsive. Ninety-four neurons in the SFO were tested for a response to electrical stimulation of the DR or hemorrhage. Electrical stimulation of the DR caused orthodromic excitation (19%) or inhibition (5%) of the activity of SFO neurons. In 14 of 18 SFO neurons that displayed the excitation to the stimulation of the DR, hemorrhage (30 to 50 mm Hg suppression in mean arterial pressure) produced an increase of their discharge, while the stimulus was without effect in the remaining neurons responsive to the stimulation of the DR. The effects of hemorrhage on serotonin (5-HT) release in the region of the SFO were examined using intracerebral microdialysis techniques. Hemorrhage significantly increased 5-HT and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) concentrations in the region of the SFO. The present data suggest that the serotonergic pathways from the DR to the SFO may relay activation of the peripheral baroreceptors to SFO neurons which result in enhanced excitability, indicating the involvement of the pathways in the regulation of cardiovascular function.     

Responses of subfornical organ neurons projecting to the hyphotalamic paraventricular nucleus to hemorrhage

The activity of subfornical organ (SFO) neurons that were antidromically identified by electrical stimulation of the rat hypothalamic paraventricular nucleus (PVN) was tested for a response to microiontophoretic application of angiotensin II (ANG II) or hemorrhage (10 ml/kg b.v.t.). Microiontophoretically (MIPh) applied ANG II caused an increased excitability in 24 out of 28 neurons tested and the excitation was blocked by MIPh-applied saralasin (Sar), a specific ANG II antagonist. Of these neurons that responded to ANG II, 14 displayed an increase in neuronal firing in response to hemorrhage, while 10 were unresponsive. The excitatory response to hemorrhage in 5 out of 14 neurons tested was prevented by MIPh-applied Sar, whereas the response of the remaining neurons was not affected. These results show that part of SFO neurons projecting to the PVN may receive neural inputs from the peripheral baroreceptors, and suggest that the inputs may be partially attributable to the involvement of central angiotensinergic circuits.     

GABAergic neurotransmission at the nucleus tractus solitarii in the suppression of reflex bradycardia by parabrachial nucleus.

We investigated the role of GABAergic neurotransmission at the nucleus tractus solitarii (NTS) in the suppression of cardiac baroreceptor reflex (BRR) response induced by parabrachial nucleus (PBN) complex in adult Sprague-Dawley rats maintained under pentobarbital anesthesia. Based on in vivo microdialysis coupled with high-performance liquid chromatography-fluorescence detection for gamma-aminobutyric acid (GABA), we found that electrical stimulation of the ventrolateral regions and Koelliker-Fuse (KF) subnucleus of PBN complex resulted in a site-specific increase in GABA concentration in the dialysate collected from the NTS. The temporal increase in extracellular GABA concentration in the NTS coincided with the time course of PBN-induced cardiac BRR inhibition. In addition, the PBN-induced cardiac BRR suppression was reversed by microinjection bilaterally into the NTS of a GABA(A) receptor antagonist, bicuculline methiodide (5 pmol), or a GABA(B) receptor antagonist, 2-OH saclofen (500 pmol). Blockade of neuronal activity in the ventrolateral regions and KF subnucleus of PBN complex with lidocaine (5%) elicited an enhancement of the same reflex response. The time course of this facilitatory effect of lidocaine correlated positively with the temporal decrease in extracellular GABA concentration in the NTS. Anatomically, Fast Blue-labeled neurons were identified in the same subnuclei of the PBN complex after microinjection of the retrograde transport tracer into the NTS. Some of these Fast Blue-labeled neurons were also immunoreactive to glutamic acid decarboxylase. These results suggest that a direct GABAergic descending projection from the KF subnucleus and surrounding areas of the PBN complex to the NTS may inhibit cardiac BRR response by activating GABA(A) and GABA(B) receptors at the NTS.     

Somatic nociception activates NK1 receptors in the nucleus tractus solitarii to attenuate the baroreceptor cardiac reflex

There is limited information regarding the integration of visceral and somatic afferents within the nucleus of the solitary tract (NTS). We studied the interaction of nociceptive and baroreceptive inputs in this nucleus in an in situ arterially perfused, un-anaesthetized decerebrate preparation of rat. At the systemic level, the gain of the cardiac component of the baroreceptor reflex was attenuated significantly by noxious mechanical stimulation of a forepaw. This baroreceptor reflex depression was mimicked by NTS microinjection of substance P and antagonized by microinjection of either bicuculline (a GABAA receptor antagonist) or a neurokinin type 1 (NK1) receptor antagonist (CP-99994). The substance P effect was also blocked by a bilateral microinjection of bicuculline, at a dose that was without effect on basal baroreceptor reflex gain. Baroreceptive NTS neurons were defined by their excitatory response following increases in pressure within the ipsilateral carotid sinus. In 27 of 34 neurons the number of evoked spikes from baroreceptor stimulation was reduced significantly by concomitant electrical stimulation of the brachial nerve (P < 0.01). Furthermore, the attenuation of baroreceptor inputs to NTS neurons by brachial nerve stimulation was prevented by pressure-ejection of bicuculline from a multi-barrelled microelectrode (n = 8). In a separate population of 17 of 45 cells tested, brachial nerve stimulation evoked an excitatory response that was antagonized by blockade of NK1 receptors. We conclude that nociceptive afferents activate NK1 receptors, which in turn excite GABAergic interneurons impinging on cells mediating the cardiac component of the baroreceptor reflex.     

Membrane and synaptic properties of nucleus tractus solitarius neurons projecting to the caudal ventrolateral medulla

Projections from the nucleus of tractus solitarius (NTS) to the caudal ventrolateral medulla (CVLM) are important in mediating autonomic reflexes. However, little is known about the cellular properties of the CVLM-projecting NTS neurons. In this study, the CVLM-projecting NTS neurons were retrogradely labeled by fluorescent microspheres injected into the CVLM. Whole cell voltage- and current-clamp recordings were performed on labeled NTS neurons in coronal brainstem slices. Compared with unlabeled neurons, the labeled NTS neurons had more depolarized resting membrane potentials, larger input resistance, and higher firing activity in response to depolarizing currents. Bath application of an ionotropic glutamate receptor antagonist kynurenic acid and a non-NMDA receptor antagonist CNQX significantly decreased the firing activity in the majority of labeled NTS neurons. In contrast, an NMDA receptor antagonist AP5 failed to alter the firing activity in labeled neurons tested. While the glycine receptor antagonist strychnine had no effect on the firing activity, blockade of GABA(A)receptors with bicuculline significantly increased the firing rate in the majority of labeled NTS neurons. Furthermore, CNQX blocked the majority of spontaneous excitatory postsynaptic currents (EPSCs) and evoked EPSCs elicited by stimulation of the tractus solitarius. The residual spontaneous and evoked EPSCs were abolished by the nicotinic receptor antagonist mecamylamine and the purinergic P2X receptor antagonist iso-PPADS. Finally, while bicuculline completely blocked the miniature inhibitory postsynaptic currents (IPSCs), the spontaneous and evoked IPSCs were abolished by a combination of bicuculline and strychnine in labeled NTS neurons. Collectively, these data suggest that the CVLM-projecting neurons are a population of neurons with distinctive membrane properties.     

Lateral hypothalamic area stimulation excites neurons in the region of the subfornical organ with efferent projections to the hypothalamic paraventricular nucleus in the rat

Fifteen neurons in the region of the subfornical organ (SFO) were antidromically activated by electrical stimulation of the paraventricular nucleus (PVN) in the rat. Electrical stimulation of the lateral hypothalamic area (LHA) excited the activity of 9 of the identified units, but did not affect the remaining units. The excitatory response of the identified units was blocked by microiontophoretically (MIPh) applied saralasin (Sar), an angiotensin II (ANGII) antagonist, but not by atropine (Atr), a muscarinic antagonist. These results suggest that the LHA has an excitatory influence on the activity of neurons in the region of the SFO with efferent projections to the PVN and that the influence may be mediated by ANGII receptors.     

Convergence of somatosensory and baroreceptive inputs onto parabrachio-subfornical organ neurons in the rat: an electrophysiological study.

Electrophysiological characteristics were described for neurons of the parabrachial nucleus (PBN) which receive baroreceptive and somatosensory inputs in the rat. Following focal electrical stimulation in the ipsilateral caudal nucleus of the tractus solitarii (NTS), the firing rates of these neurons were increased in 94 (55.6%), and decreased in 38 (22.5%). Fifty-three (54.5%) of 97 PBN neurons tested were excited, and 11 (11.3%) inhibited in response to contralateral common peroneal nerve (CPN) stimulation. Of these neurons, 52 were found to respond to both caudal NTS and CPN stimulation. Effects of subfornical organ (SFO) stimulation were also examined in 151 of 169 parabrachial neurons. 13 (7.7%) were activated antidromically and were located in the lateral division of the PBN, while 34 (22.5%) were affected orthodromically. Stimulation of the caudal NTS resulted in both a fall in the heart rate and changes of PBN neuronal firing rates. Similar effects were elicited by activating peripheral baroreceptors by the administration of phenylephrine, an alpha-adrenergic agonist. These results strongly indicate that: (1) the lateral PBN is involved in central cardiovascular control; (2) somatosensory and baroreceptive messages may converge onto some PBN neurons; and (3) some PBN neurons may relay baroreceptive information from the caudal NTS to the SFO.     

Descending noradrenergic pathways involved in the A5 depressor response

The objective of the present study was to analyze the anatomical basis of the A₅ depressor response and to test if the putative neurotransmitter noradrenaline is involved in the response. Two approaches were used; one was neuroanatomical and the other was pharmacological. First, the retrograde transport method in which two fluorescent markers (Fast blue and rhodamine microspheres) was used in combination with the indirect immunofluorescence technique to establish that A₅ catecholamine neurons project to both the spinal cord and the region of the nucleus tractus solitarii (NTS). Second, we analyzed the effects of 6-hydroxydopamine (6-OHDA) lesions of the spinal cord and/or NTS area on the A₅ depressor response. This response was elicited by a 80-nl microinjection ofl-glutamate (500 mM) into the A₅ region in pentobarbital anesthetized rats; it was characterized by a decrease in blood pressure and heart rate. After destruction of various noradrenergic terminal fields we have found that (1) intraspinal injections of 6-OHDA caused a 30% reduction in the blood pressure component of the A₅ depressor response and a transient depression of the bradycardic response. This result suggests that only a small portion of the A₅ depressor response depends on the descending A₅ spinal pathway. (2) Injections of 6-OHDA into the NTS region caused a transient depression of the A5 depressor response, and by 7–14 days postinjection, the response returned to normal. (3) After combined 6-OHDA injections into the spinal cord and NTS area, the blood pressure and heart rate components of the A₅ depressor response were reduced to 80% of the control level at 3 days postinjection. By 14 days, even with severe depletion of noradrenaline in the spinal cord (96%) and a moderate depletion of noradrenaline in the NTS (50%), the A₅ response was restored to about 80% of its original magnitude, suggesting some type of functional recovery occurs in this system. Third, the blood pressure decrease elicited byl-glutamate stimulation of the A₅ cell group was unaffected by pharmacological blockade of the heart. In addition, this response appeared to be normal in rats that had both their autonomic supply to the heart blocked pharmacologically and their spinal cord noradrenaline levels depleted (14 days after intraspinal 6-OHDA injections). These data suggest that the major A₅ depressor response operates mainly by inhibition of the sympathetic outflow involved in control of total peripheral resistance and that this system is controlled by a descending spinal pathway which probably does not use noradrenaline as a neurotransmitter.     

Gamma aminobutyric acid regulates glucosensitive neuropeptide Y neurons in arcuate nucleus via A/B receptors

Gamma aminobutyric acid (GABA) is localized in neuropeptide Y (NPY) neurons of the hypothalamic arcuate nucleus (ARC). We examined regulation of ARC NPY neurons by GABA. Light and electron microscopic immunohistochemistry confirmed that GABA-containing nerve terminals contacted NPY-containing neurons in the ARC. Lowering glucose (1 mM) increased cytosolic Ca2+ concentration ([Ca2+]i) in isolated ARC neurons that were immunoreactive to NPY. The [Ca2+]i increases were inhibited by GABA, the gamma-aminobutyric acid type A receptor (GABAA) agonist muscimol and the gamma-aminobutyric acid type B receptor (GABAB) agonist baclofen. Neither the GABAA antagonist bicuculline nor the GABAB antagonist CGP35348 counteracted the GABA inhibition when applied alone, but did so when applied together. These results indicate that GABA regulates ARC glucose-sensitive NPY neurons via GABAA and GABAB receptors, which could function to attenuate the orexigenic NPY pathway when it is not beneficial.     

Connections of neurons in the region of the nucleus tractus solitarius with the hypothalamic paraventricular nucleus: Their possible involvement in neural control of the cardiovascular system in rats

Extracellular recordings were made from 607 spontaneously firing neurons within the nucleus tractus solitarius (NTS) and its vicinity in urethane-anesthetized male rats. Following electrical stimulation of the hypothalamic paraventricular nucleus (PVN) area, 21% of the neurons were orthodromically excited, 6% were inhibited and 2.5% were antidromically activated. The antidromic spike latencies were 22-64 ms. Among those orthodromically responding neurons, 81 neurons were tested by pressure pulse stimulation of the isolated carotid sinus. The pressure stimulation produced excitation in 7 and inhibition in 13 neurons. Of the 8 tested neurons which were antidromically activated, one neuron was excited and another neuron inhibited by the pressure pulse stimulation. These results provide electrophysiological evidence for reciprocal connections between neurons in the NTS region and the PVN, and give support to the hypothesis that the PVN is involved in the neural control of the cardiovascular system.     

GABA-mediated inhibition of locus coeruleus from the dorsomedial rostral medulla

Recent anatomic studies in our laboratory (Aston-Jones et al., 1986) identified the nucleus prepositus hypoglossi (PrH) in the dorsomedial medulla as a major afferent of the locus coeruleus (LC). In the present studies, the influence of projections from PrH to LC was assessed in anesthetized rats. Focal electrical stimulation of PrH inhibited the spontaneous discharge of 42 of 47 LC neurons; the latency to onset of such inhibition was 19.8 +/- 2.5 msec and its duration was 172.4 +/- 10.4 msec. PrH-evoked inhibition of LC neurons was unaffected by administration of the opiate receptor antagonist naloxone or the alpha 2-receptor antagonist idazoxan but was substantially reduced by systemic picrotoxin, an antagonist of GABA. The GABAA receptor antagonist bicuculline methiodide blocked the inhibition from PrH, whether applied by local microinfusion or iontophoresis into the LC. These results lead us to propose that PrH provides a direct inhibitory synaptic input to LC, for which GABA is the likely transmitter.     

Effects of electrical stimulation of the pontine A5 cell group on blood pressure and heart rate in the rabbit

The effects of electrical stimulation of the A5 noradrenergic cell group of the ventrolateral pons was assessed in rabbits. Stimulation administered through either concentric bipolar or monopolar electrodes produced current-intensity related increases in mean arterial pressure (MAP). Decreases in heart rate (HR) accompanied the increases in MAP, but were essentially eliminated by bilateral vagotomy or destruction of the nucleus and tractus solitarii (NTS), thereby indicating that the HR decelerations were secondary to activation of baroreceptor reflexes. Neither vagotomy nor midcollicular section of the brainstem altered the MAP response to A5 stimulation. Bilateral destruction of the NTS slightly enhanced the response. Several rabbits received microinjections of 6-hydroxydopamine (6-OHDA) into the A5 region 2 weeks before the experiment. Other rabbits received vehicle injections and served as control subjects for the non-specific effects of the 6-OHDA injections. 6-OHDA injections, but not vehicle injections, prevented the vasopressor effects of A5 stimulation. However, stimulation of the A1 noradrenergic nucleus of the ventrolateral medulla produced decreases in MAP in rabbits given prior microinjections of 6-OHDA into A5. These observations are interpreted to indicate that the 6-OHDA injections produced neurotoxic effects which were relatively restricted to the A5 region. Furthermore, the data from all of these experiments are interpreted as indicating that the vasopressor effects observed as a consequence of electrical stimulation of A5 are due to excitation of the noradrenaline-containing neuron cell bodies of this region and that this effect is mediated via pathways arising from this region and terminating in the intermediolateral cell column of the spinal cord.     

Decreased effectiveness of GABA-mediated inhibition in the inferior colliculus of the genetically epilepsy-prone rat

The inferior colliculus (IC) is a critical site for induction of audiogenic seizures in the genetically epilepsy-prone rat (GEPR). Abnormal response properties observed in inferior colliculus neurons of that strain include a high incidence of onset-offset responses which may be a form of afterdischarge. These response abnormalities may involve altered actions of neurotransmitters in that region of the brain. GABA is implicated as a transmitter in endogenous sound-induced inhibition in the inferior colliculus. Endogenous inhibition and the actions of agents that affect GABA receptors were examined in inferior colliculus neurons in epileptic and normal rats. The iontophoretic dose (current) of GABA required to suppress neuronal firing in the epilepsy-prone rat was significantly greater than that required in neurons of the normal rat. A form of endogenous (binaural) inhibition in inferior colliculus neurons, which is proposed to be GABA-mediated, was also significantly reduced in the epilepsy-prone rat as compared with the normal rat. A benzodiazepine (flurazepam) which enhances the action of GABA in many brain sites including the inferior colliculus was significantly less effective than normal when applied iontophoretically onto the same neurons of the epilepsy-prone rat. The GABAA antagonist, bicuculline, which blocks the effect of GABA on inferior colliculus neurons, frequently induced the onset-offset response in neurons not previously exhibiting this pattern. These findings suggest that the reduction of GABA-mediated inhibition in the genetically epilepsy-prone rat may result in the increased incidence of afterdischarges in neurons of the inferior colliculus and may serve as an important mechanism of epileptogenesis in audiogenic seizures in this genetic form of epilepsy.     

Estrogen decreases the responsiveness of subfornical organ neurons to angiotensinergic neural inputs from the lateral hypothalamic area in the female rat

Twenty-eight subfornical organ (SFO) neurons in ovariectomized (OVX) female rats that were treated with propylene glycol (PG) vehicle and 26 SFO neurons in OVX female rats that were treated with estrogen benzoate (EB) were antidromically activated by electrical stimulation of the hypothalamic paraventricular nucleus (PVN) under urethane anesthesia. No significant differences were observed between the PG-treated and EB-treated OVX animals in the latency, conduction velocity, or threshold of antidromic activation. The mean spontaneous discharge rate was significantly lower in the EB-treated than in the PG-treated OVX animals. In both groups, the activity of the majority (86% in the PG-treated animals and 88% in the EB-treated animals) of identified SFO neurons were activated by microiontophoretic application of angiotensin II (ANG II). Electrical stimulation of the lateral hypothalamic area (LHA) increased the excitability of these ANG II-sensitive SFO neurons (58% in the PG-treated animals and 52% in the EB-treated animals). The excitatory response to either ANG II or LHA stimulation was blocked by microiontophoretic application of the ANG II antagonist saralasin (Sar), suggesting that the excitatory response to LHA stimulation may be mediated by angiotensinergic LHA projections to the SFO. The magnitude of excitatory response to either ANG II or the LHA stimulation was much greater in the PG-treated than in the EB-treated animals. These results suggest that estrogen decreases the responsiveness of SFO neurons projecting to the PVN to angiotensinergic inputs from the LHA.     

Regulation of Limbic Motor Seizures by GABA and Glutamate Transmission in Nucleus Tractus Solitarius

PURPOSE: The nucleus of the solitary tract (NTS) is a primary site at which vagal afferents terminate. Because afferent vagal nerve stimulation has been demonstrated to have anticonvulsant effects, it is likely that changes in synaptic transmission in the NTS can regulate seizure susceptibility. We tested this hypothesis by examining the influence of gamma-aminobutyric acid (GABA) ergic and glutamatergic transmission in the NTS on seizures evoked by systemic and focal bicuculline and systemic pentylenetetrazol (PTZ) in rats. METHODS: Muscimol (256 pmol), a GABA(A)-receptor agonist, bicuculline methiodide (177 pmol), a GABA(A)-receptor antagonist, kynurenate (634 pmol), a glutamate-receptor antagonist, or lidocaine (100 nl; 5%), a local anesthetic, was microinjected into the mediocaudal (m)NTS. Ten minutes later, seizure activity was induced by either a focal microinfusion of bicuculline methiodide (177 pmol) into the rostral piriform cortex, systemic PTZ (50 mg/kg, i.p.), or systemic bicuculline (0.35 mg/kg, i.v.). RESULTS: Muscimol in mNTS (but not in adjacent regions of NTS) attenuated seizures in all seizure models tested, whereas bicuculline methiodide into mNTS did not alter seizure responses. Kynurenate infusions into mNTS significantly reduced the severity of seizures evoked both systemically and focally. Anticonvulsant effects also were obtained with lidocaine application into the same region of mNTS. Unilateral injections were sufficient to afford seizure protection. CONCLUSIONS: Our results demonstrate that an increase in GABA transmission or a decrease in glutamate transmission in the rat mNTS reduces susceptibility to limbic motor seizures. This suggests that inhibition of mNTS outputs enhances seizure resistance in the forebrain and provides a potential mechanism for the seizure protection obtained with vagal stimulation.     

Tooth-pulp-evoked rostral spinal trigeminal neuronal excitation is attenuated by the activation of 5-HT3 receptors via GABAergic interneurons in the rat

The effect of iontophoretic application of the 5-HT3 receptor agonist, phenylbiguanide (PBG), on the excitation of the trigeminal spinal nucleus oralis (TSNO) neurons to tooth-pulp (TP) stimulation was examined. The PBG application inhibited the TP-evoked TSNO neuronal excitation, and this inhibition was completely blocked by co-application of a GABAA receptor antagonist, bicuculline. The results suggest that the activation of 5-HT3 receptors elicits GABA release in the TSNO.     

GABAA receptor modulation of trigeminovascular nociceptive neurotransmission by midazolam is antagonized by flumazenil

Studies of the pharmacology of trigeminocervical neurons with input from intracranial pain-producing structures have enhanced the understanding of the basic neurobiology of primary headache, such as migraine. Clinical observations of the treatment of migraine with medicines acting at the gamma-aminobutyric acid (GABA) GABAA receptor have lead to studies of their effects on models of trigeminovascular nociception. Extracellular recordings were made from neurons in the trigeminocervical complex activated by supramaximal electrical stimulation of superior sagittal sinus (SSS) in the cat. Intravenous administration of the benzodiazepine receptor agonist midazolam, resulted in a dose-dependent inhibition of superior sagittal sinus evoked trigeminocervical nucleus activity. The inhibition at 50 microg/kg midazolam was 65+/-11% compared to the baseline response (n=11). Intravenous administration of the benzodiazepine receptor antagonist flumazenil, resulted in a dose-dependent recovery of superior sagittal sinus evoked trigeminocervical nucleus activity. At a dose of 50 microg/kg, there was a 64+/-5% recovery (n=6). The data demonstrate a potent, reproducible effect of facilitation of GABA transmission at the GABAA receptor that results in inhibition of trigeminovascular nociceptive transmission. These data are consistent with the useful clinical effects reported with compounds that can augment GABAergic transmission in the central nervous system (CNS).