Activity Of Nucleus Tractus Solitarius Neurons Research Articles

Synaptic mechanisms underlying the elevated sympathetic outflow in fructose-induced hypertension.

Metabolic syndrome is associated with cardiovascular dysfunction, including elevated sympathetic outflow. However, the underlying brain mechanisms are unclear. The nucleus tractus solitarius (NTS) critically regulates autonomic reflexes related to cardiovascular function and contains neurons projecting to the caudal ventrolateral medulla (CVLM). Nitric oxide (NO) is a diffusible free-radical messenger in the vascular, immune, and nervous systems. In this study, we determine if NO in the NTS is involved in the synaptic plasticity underlying the elevated sympathetic outflow in fructose-induced hypertension. We retrogradely labeled CVLM-projecting NTS neurons through the injection of FluoSpheres into the CVLM in a fructose-fed rat model to determine the cellular mechanism involved in increased sympathetic outflow. Fructose feeding increased the blood pressure and glucose levels, which represent metabolic syndrome. We found that fructose feeding reduces the NO precursor L-arginine-induced increase in the firing activity of CVLM-projecting NTS neurons. Furthermore, fructose feeding reduces the L-arginine-induced increase in presynaptic spontaneous glutamatergic synaptic inputs to NTS neurons, while NO donor DEA/NO produces an increase in glutamatergic synaptic inputs in fructose-fed rats similar to that in vehicle-treated rats. In addition, fructose feeding reduces the NO-induced depressor response and sympathoinhibition. These data suggested that fructose feeding reduced NO production and, thus, the subsequent NO-induced glutamate releases in the NTS and depressor response. The findings of this study provide new insights into the central mechanisms involved in the neural control of cardiovascular and autonomic functions in the NTS in metabolic syndrome.

Systemic Glucose Regulation by a Hindbrain Inhibitory Circuit in a Mouse Model of Type 1 Diabetes

Introduction: Previous work showed that increasing the electrical activity of inhibitory neurons in the dorsal vagal complex (DVC) is sufficient to increase whole-body glucose concentration in normoglycemic mice. Here we tested the hypothesis that deactivating GABAergic neurons in the dorsal hindbrain of hyperglycemic mice decreases synaptic inhibition of parasympathetic motor neurons in the dorsal motor nucleus of the vagus (DMV) and reduces systemic glucose levels. Methods: Chemogenetic activation or inactivation of GABAergic neurons in the nucleus tractus solitarius (NTS) was used to assess effects of modulating parasympathetic output on blood glucose concentration in normoglycemic and hyperglycemic mice. Patch-clamp electrophysiology in vitro was used to assess cellular effects of chemogenetic manipulation of NTS GABA neurons. Results: Chemogenetic activation of GABAergic NTS neurons in normoglycemic mice increased their action potential firing, resulting in increased inhibitory synaptic input to DMV motor neurons and elevated blood glucose concentration. Deactivation of GABAergic DVC neurons in normoglycemic mice altered their electrical activity but did not alter systemic glucose levels. Conversely, stimulation of GABAergic DVC neurons in mice that were hyperglycemic subsequent to treatment with streptozotocin changed their electrical activity but did not alter whole-body glucose concentration, while deactivation of this inhibitory circuit significantly decreased circulating glucose concentration. Peripheral administration of a brain impermeant muscarinic acetylcholine receptor antagonist abolished these effects. Conclusion: Disinhibiting vagal motor neurons decreases hyperglycemia in a mouse model of type 1 diabetes. This inhibitory brainstem circuit emerges as a key parasympathetic regulator of whole-body glucose homeostasis that undergoes functional plasticity in hyperglycemic conditions.

The carotid body detects circulating tumor necrosis factor-alpha to activate a sympathetic anti-inflammatory reflex

Recent evidence has suggested that the carotid bodies might act as immunological sensors, detecting pro-inflammatory mediators and signalling to the central nervous system, which, in turn, orchestrates autonomic responses. Here, we confirmed that the TNF-α receptor type I is expressed in the carotid bodies of rats. The systemic administration of TNF-α increased carotid body afferent discharge and activated glutamatergic neurons in the nucleus tractus solitarius (NTS) that project to the rostral ventrolateral medulla (RVLM), where many pre-sympathetic neurons reside. The activation of these neurons was accompanied by an increase in splanchnic sympathetic nerve activity. Carotid body ablation blunted the TNF-α-induced activation of RVLM-projecting NTS neurons and the increase in splanchnic sympathetic nerve activity. Finally, plasma and spleen levels of cytokines after TNF-α administration were higher in rats subjected to either carotid body ablation or splanchnic sympathetic denervation. Collectively, our findings indicate that the carotid body detects circulating TNF-α to activate a counteracting sympathetic anti-inflammatory mechanism.

Fibroblast Growth Factor 19 Increases the Excitability of Pre-Motor Glutamatergic Dorsal Vagal Complex Neurons From Hyperglycemic Mice.

Intracerebroventricular administration of the protein hormone fibroblast growth factor 19 (FGF19) to the hindbrain produces potent antidiabetic effects in hyperglycemic mice that are likely mediated through a vagal parasympathetic mechanism. FGF19 increases the synaptic excitability of parasympathetic motor neurons in the dorsal motor nucleus of the vagus (DMV) from hyperglycemic, but not normoglycemic, mice but the source of this synaptic input is unknown. Neurons in the area postrema (AP) and nucleus tractus solitarius (NTS) express high levels of FGF receptors and exert glutamatergic control over the DMV. This study tested the hypothesis that FGF19 increases glutamate release in the DMV by increasing the activity of glutamatergic AP and NTS neurons in hyperglycemic mice. Glutamate photoactivation experiments confirmed that FGF19 increases synaptic glutamate release from AP and NTS neurons that connect to the DMV in hyperglycemic, but not normoglycemic mice. Contrary to expectations, FGF19 produced a mixed effect on intrinsic membrane properties in the NTS with a trend towards inhibition, suggesting that another mechanism was responsible for the observed effects on glutamate release in the DMV. Consistent with the hypothesis, FGF19 increased action potential-dependent glutamate release in the NTS in hyperglycemic mice only. Finally, glutamate photoactivation experiments confirmed that FGF19 increases the activity of glutamatergic AP neurons that project to the NTS in hyperglycemic mice. Together, these results support the hypothesis that FGF19 increases glutamate release from AP and NTS neurons that project to the DMV in hyperglycemic mice. FGF19 therefore modifies the local vago-vagal reflex circuitry at several points. Additionally, since the AP and NTS communicate with several other metabolic regulatory nuclei in the brain, FGF19 in the hindbrain may alter neuroendocrine and behavioral aspects of metabolism, in addition to changes in parasympathetic output.

RETRACTED: Sustained Hypoxia Reduces GABAergic Modulation on NTS Neurons Sending Projections to Ventral Medulla of Rats

Peripheral chemoreflex is activated during short-term sustained hypoxia (SH), and the first synapse of these afferents is located in Nucleus Tractus Solitarius(NTS). NTS neurons projecting to the ventral lateral medulla (NTS-VLM) are part of the respiratory pathways of the chemoreflex. SH increases the magnitude of basal respiratory parameters in rats from Wistar-Hannover strain. In this study, we hypothesized that the observed changes in the respiratory pattern in response to SH were due to changes in the GABAergic modulation of the synaptic transmission of NTS-VLM neurons. We used an electrophysiological approach to record the synaptic activity of NTS neurons labeled with a retrograde tracer previously microinjected into VLM of Wistar-Hannover rats submitted to 24 h SH. The data are showing that: (a) the amplitude of evoked inhibitory currents in NTS-VLM neurons of SH rats was reduced and not accompanied by any change in rise-time and decay-time; (b) the 1/CV2 and the number of failures in response to evoked currents were also affected by SH; (c) the frequency of spontaneous inhibitory currents was reduced by SH without changes in amplitude and half-width. These effects of SH were observed in NTS-VLM neurons located in caudal and intermediate NTS, but not in NTS-VLM neurons located in the rostral NTS. We conclude that SH causes a reduction in inhibitory modulation onto NTS-VLM neurons by pre-synaptic mechanisms, which may contribute to the observed changes in the respiratory pattern of Wistar-Hannover rats submitted to SH.

PVN‐projecting NTS Neurons is Involved in Sympathetic Long‐Term Facilitation After Acute Intermittent Optogenetic Stimulation

Caudal Nucleus Tractus Solitarius (NTS) neurons receive inputs from cardiorespiratory visceral afferents and send projections to the paraventricular nucleus of the hypothalamus (PVN). Acute intermittent optogenetic (AIO) stimulation of caudal NTS induces sympathetic long‐term facilitation (LTF), a progressive increase of sympathetic nerve activity. The contribution of PVN‐projecting NTS neurons to sympathetic LTF is unknown. This study used an intersectional viral approach to test the effects of acute intermittent optogenetic (AIO) stimulation of PVN‐projecting NTS neurons in sympathetic LTF. Adult male Sprague‐Dawley rats were anesthetized with isoflurane and injected rAAV2/Ef1a‐DIO‐hchR2(H134R)‐mCherry in the NTS and a retrograde AAV9.hSyn.HI.eGFP‐cre.WPRE.SV40 in the PVN four week prior to LTF experiments. This method was used to induce the CRE dependent expression of channel rhodopsin in PVN‐projecting NTS neurons. To measure LTF, we recorded splanchnic sympathetic nerve activity (SSNA), arterial pressure, and heart rate (HR) in anesthetized (urethane (800mg/kg) and α‐chloralose (80 mg/kg) cocktail delivered by intraperitoneal injection), paralyzed with gallamine triethiodide (20 mg/ml, 0.25 ml/h, iv) and mechanically ventilated adult rats. The caudal NTS was stimulated to induce AIO with a train of 10 stimuli each consisting of pulse of light of 10‐ms duration delivered at 20 Hz for 1 min every 6 min. Twenty minutes after AIO, SSNA amplitude increased by 25.36 ± 24.16% (n=3), and mean arterial pressure (MAP) increased by 7.43 ± 12.88 mmHg (n=3). During AIO induction, the acute SSNA and MAP varied with some rats demonstrating increases (n=2) or decreases (n=1) of SSNA, MAP, and HR. In other animals (n=3) where AIO decreases SSNA (‐58.96 ± 36.18%) and MAP (‐14.37 ± 8.57 mmHg), we tested the possible contribution of the baroreceptors in these decreases by bilateral muscimol (1–10 pmol/100nL) injections in the Caudal Ventrolateral Medulla (CVLM). AIO stimulation after inhibition of CVLM caused an increase in SSNA (1.293 ± 17.65%) and smaller decrease in MAP (−2.46± 6.2 mmHg). These results suggest that the splanchnic sympathetic LTF induced by acute intermittent stimulation of NTS neurons might be mediated by the PVN‐projecting neurons, although the acute responses may be excitatory or inhibitory. Further study is needed to clarify the mechanisms responsible for the differences in the cardiovascular and SSNA responses to acute, single optogenetic stimulation of PVN‐projecting NTS neurons. Despite the observed differences in the acute responses, the results indicate that sympathetic LTF can be induced by the optogenetic activation of NTS neurons that project to PVN.Support or Funding InformationNIH Grant P01 HL088052

A hindbrain inhibitory microcircuit mediates vagally-coordinated glucose regulation

Neurons in the brainstem dorsal vagal complex integrate neural and humoral signals to coordinate autonomic output to viscera that regulate a variety of physiological functions, but how this circuitry regulates metabolism is murky. We tested the hypothesis that premotor, GABAergic neurons in the nucleus tractus solitarius (NTS) form a hindbrain micro-circuit with preganglionic parasympathetic motorneurons of the dorsal motor nucleus of the vagus (DMV) that is capable of modulating systemic blood glucose concentration. In vitro, neuronal activation or inhibition using either excitatory or inhibitory designer receptor exclusively activated by designer drugs (DREADDs) constructs expressed in GABAergic NTS neurons increased or decreased, respectively, action potential firing of GABAergic NTS neurons and downstream synaptic inhibition of the DMV. In vivo, DREADD-mediated activation of GABAergic NTS neurons increased systemic blood glucose concentration, whereas DREADD-mediated silencing of these neurons was without effect. The DREADD-induced hyperglycemia was abolished by blocking peripheral muscarinic receptors, consistent with the hypothesis that altered parasympathetic drive mediated the response. This effect was paralleled by elevated serum glucagon and hepatic phosphoenolpyruvate carboxykinase 1 (PEPCK1) expression, without affecting insulin levels or muscle metabolism. Activity in a hindbrain inhibitory microcircuit is sufficient to modulate systemic glucose concentration, independent of insulin secretion or utilization.

Colitis-induced alterations in response properties of visceral nociceptive neurons in the rat caudal medulla oblongata and their modulation by 5-HT3 receptor blockade

There is considerable clinical and experimental evidence that intestinal inflammation is associated with altered visceral nociceptive processing in the spinal cord and brain, but the underlying neuronal mechanisms, especially acting at the supraspinal level, remain unclear. Considering that the caudal ventrolateral medulla (CVLM) and the nucleus tractus solitarius (NTS) are the first sites for supraspinal processing of visceral pain signals, in the present study we evaluated the experimental colitis-induced changes in response properties of CVLM and NTS medullary neurons to noxious colorectal distension (CRD) in urethane-anesthetized adult male Wistar rats. To determine if gut inflammation alters the 5-HT3 receptor-dependent modulation of visceral pain-related CVLM and NTS cells, we examined the effects of intravenously administered selective 5-HT3 antagonist granisetron on ongoing and CRD-evoked activity of CVLM and NTS neurons in healthy control and colitic animals. In the absence of colonic pathology, the CVLM neurons were more excited by noxious CRD that the NTS cells, which demonstrated a greater tendency to be inhibited by the stimulation. The difference was eliminated after the development of colitis due to the increase in the proportion of CRD-excited neurons in both medullary regions associated with enhanced magnitude of the neuronal nociceptive responses. Intravenous granisetron (1 or 2 mg/kg) produced the dose-dependent suppression of the ongoing and evoked firing of CRD-excited cells within both the CVLM and NTS in normal conditions as well as was able to substantially reduce excitability of the caudal medullary neurons in the presence of colonic inflammation, arguing for the potential efficacy of the 5-HT3 receptor blockade with granisetron against both acute and inflammatory abdominal pain. Taken together, the data obtained can contribute to a deeper understanding of supraspinal serotonergic mechanisms responsible for the persistence of visceral hypersensitivity and hyperalgesia triggered by colonic inflammation.

Functional Neuroplasticity in the Nucleus Tractus Solitarius and Increased Risk of Sudden Death in Mice with Acquired Temporal Lobe Epilepsy.

Sudden unexpected death in epilepsy (SUDEP) is the leading cause of death in individuals with refractory acquired epilepsy. Cardiorespiratory failure is the most likely cause in most cases, and central autonomic dysfunction has been implicated as a contributing factor to SUDEP. Neurons of the nucleus tractus solitarius (NTS) in the brainstem vagal complex receive and integrate vagally mediated information regarding cardiorespiratory and other autonomic functions, and GABAergic inhibitory NTS neurons play an essential role in modulating autonomic output. We assessed the activity of GABAergic NTS neurons as a function of epilepsy development in the pilocarpine-induced status epilepticus (SE) model of temporal lobe epilepsy (TLE). Compared with age-matched controls, mice that survived SE had significantly lower survival rates by 150 d post-SE. GABAergic NTS neurons from mice that survived SE displayed a glutamate-dependent increase in spontaneous action potential firing rate by 12 wks post-SE. Increased spontaneous EPSC frequency was also detected, but vagal afferent synaptic release properties were unaltered, suggesting that an increase in glutamate release from central neurons developed in the NTS after SE. Our results indicate that long-term changes in glutamate release and activity of GABAergic neurons emerge in the NTS in association with epileptogenesis. These changes might contribute to increased risk of cardiorespiratory dysfunction and sudden death in this model of TLE.

GABA in nucleus tractus solitarius participates in electroacupuncture modulation of cardiopulmonary bradycardia reflex.

Phenylbiguanide (PBG) stimulates cardiopulmonary receptors and cardiovascular reflex responses, including decreases in blood pressure and heart rate mediated by the brain stem parasympathetic cardiac neurons in the nucleus ambiguus and nucleus tractus solitarius (NTS). Electroacupuncture (EA) at P5-6 stimulates sensory fibers in the median nerve and modulates these reflex responses. Stimulation of median nerves reverses bradycardia through action of γ-aminobutyric acid (GABA) in the nucleus ambiguus, important in the regulation of heart rate. We do not know whether the NTS or the neurotransmitter mechanisms in this nucleus participate in these modulatory actions by acupuncture. We hypothesized that somatic nerve stimulation during EA (P5-6) modulates cardiopulmonary inhibitory responses through a GABAergic mechanism in the NTS. Anesthetized and ventilated cats were examined during either PBG or direct vagal afferent stimulation while 30 min of EA was applied at P5-6. Reflex heart rate and blood pressure responses and NTS-evoked discharge were recorded. EA reduced the PBG-induced depressor and bradycardia reflexes by 67% and 60%, respectively. Blockade of GABAA receptors in the NTS reversed EA modulation of bradycardia but not the depressor response. During EA, gabazine reversed the vagally evoked discharge activity of cardiovascular NTS neurons. EA modulated the vagal-evoked cardiovascular NTS cellular activity for 60 min. Immunohistochemistry using triple labeling showed GABA immunoreactive fibers juxtaposed to glutamatergic nucleus ambiguus-projecting NTS neurons in rats. These glutamatergic neurons expressed GABAA receptors. These findings suggest that EA inhibits PBG-evoked bradycardia and vagally evoked NTS activity through a GABAergic mechanism, likely involving glutamatergic nucleus ambiguus-projecting NTS neurons.

Endocannabinoid effects on neuronal function in the NTS of hypertensive rats (1130.3)

Previously, we showed that endocannabinoid activation of cannabinoid 1 receptors (CB1Rs) in the nucleus tractus solitarius (NTS) activates barosensitive neurons and prolongs baroreflex inhibition of renal sympathetic nerve activity in normotensive Sprague Dawley (SD) rats, but reflex responses were attenuated in spontaneously hypertensive rats (SHR). This study examined the effect of CB1Rs on NTS barosensitive neurons in SHRs to determine if the lack of reflex responses is associated with diminished effects on neuronal firing. NTS neuronal activity was recorded in the working heart brainstem preparation in SHR or SD rats, with increases in perfusion pressure used to identify barosensitive firing. To examine the effects of CB1Rs on neuronal discharge, picoejections of the CB1R agonist anandamide (AEA), antagonist SR141716, or blocker of AEA catabolism, AM404, were made through a multibarrel pipette array which contained the carbon fiber recording electrode. AEA acting at CB1Rs increased barosensitive neuronal firing in SD versus SHR rats. In addition, discharge of respiratory I neurons, identified by correlation to phrenic nerve discharge, was found to respond more to CB1R activation in SD rats versus SHRs. The data indicate that neuronal function of CB1Rs in the NTS is blunted in SHRs.Grant Funding Source: Suppoted by VA Merit Review I01 BX000481 (JLS), NSF Award IOS 0751613 (CD)

P01.45. Activated neuropathway from nucleus tractus solitarius to rostal ventrolateral medulla during electroacupuncture

Purpose Our previous studies have shown that electroacupuncture (EA) at the Jianshi-Neiguan acupoints (P5-P6, overlying the median nerve) attenuates sympathoexcitatory responses through its influence on neuronal activity in the rostral ventrolateral medulla (rVLM). Nucleus tractus solitarius (NTS) receives inputs from somatic nerve stimulation. However, there is no information on the activation of NTS neurons by EA at P5-P6 acupoints, which subsequently affects the rVLM. Thus, the present study evaluated neuronal activation of NTS in response to EA, with regard to their projections to the rVLM. Methods Seven to ten days after unilateral microinjection of a rodamine-conjugated microsphere retrograde tracer (100 nl) into the rVLM, rats were subjected to EA or served as a sham-operated control. EA was performed for 30 min at P5-P6 acupoints bilaterally. Results Perikarya containing the microsphere tracer were found in the NTS of both groups. Compared to controls (needle placement without electrical stimulation, n=4), c-Fos immunoreactivity and neurons double-labeled with cFos, an immediate early gene and the tracer were more frequently found in the NTS of EA-treated rats (n=5), particularly, in the medial and lateral subdivisions of caudal and intermediate NTS extension. Conclusion These results suggest that EA at P5-P6 acupoints activates NTS neurons. Furthermore, EA-activated NTS neuron can directly project to the rVLM, which is known to participate in EA-modulation of sympathetic activity.

Integrative responses of neurons in nucleus tractus solitarius to stimulation of visceral and vestibular afferents

Although the vestibular nuclei project to nucleus tractus solitarius (NTS), little is known about the effects of vestibular inputs on NTS neuronal activity. Lesions of NTS abolish vomiting elicited by a variety of triggering mechanisms, including vestibular inputs. Thus, an emetic stimulus that activates gastrointestinal (GI) receptors could alter the responses of NTS neurons to vestibular inputs. We examined in decerebrate cats the responses of NTS neurons to rotations of the body before and after the intragastric administration of the emetic compound CuSO4. The activity of > 30% NTS neurons was modulated by vestibular stimulation, with most of the responsive cells having their firing rate altered by rotations in the head‐up or head‐down directions. These responses were aligned with head position in space, as opposed to the velocity of head movements. The activity of NTS neurons with baroreceptor, pulmonary and GI inputs could be modulated by vertical plane rotations. However, injection of CuSO4 into the stomach did not alter the responses to vestibular stimulation of NTS neurons that received GI inputs, suggesting that the stimuli did not have additive effects. These findings show that the detection and processing of visceral inputs by NTS neurons can be altered in accordance with the direction of ongoing movements.

Integrative responses of neurons in nucleus tractus solitarius to visceral afferent stimulation and vestibular stimulation in vertical planes.

Anatomical studies have demonstrated that the vestibular nuclei project to nucleus tractus solitarius (NTS), but little is known about the effects of vestibular inputs on NTS neuronal activity. Furthermore, lesions of NTS abolish vomiting elicited by a variety of different triggering mechanisms, including vestibular stimulation, suggesting that emetic inputs may converge on the same NTS neurons. As such, an emetic stimulus that activates gastrointestinal (GI) receptors could alter the responses of NTS neurons to vestibular inputs. In the present study, we examined in decerebrate cats the responses of NTS neurons to rotations of the body in vertical planes before and after the intragastric administration of the emetic compound copper sulfate. The activity of more than one-third of NTS neurons was modulated by vertical vestibular stimulation, with most of the responsive cells having their firing rate altered by rotations in the head-up or head-down directions. These responses were aligned with head position in space, as opposed to the velocity of head movements. The activity of NTS neurons with baroreceptor, pulmonary, and GI inputs could be modulated by vertical plane rotations. However, injection of copper sulfate into the stomach did not alter the responses to vestibular stimulation of NTS neurons that received GI inputs, suggesting that the stimuli did not have additive effects. These findings show that the detection and processing of visceral inputs by NTS neurons can be altered in accordance with the direction of ongoing movements.

Ascorbic acid and α-tocopherol supplement starting prenatally enhances the resistance of nucleus tractus solitarius neurons to hypobaric hypoxic challenge

Hypobaric hypoxia, encountered at high altitude, could result in severe consequences. Ascorbic acid (AA) and α-tocopherol (αTC), the two readily available over-the-counter antioxidants, are known to protect nervous tissue against oxidative stress. Here we study whether AA or αTC supplement starting prenatally protects animals against hypobaric hypoxic challenge at adulthood. Expressions of c-fos and the NR1 subunit of the N-methyl-D-aspartate receptors in the nucleus tractus solitarius (NTS) subserving cardiorespiratory functions were investigated. AA and αTC supplement reduced the number of c-fos immunoreactive neurons and intensity of NR1 expression in young and adult animals under normoxia. The treatment, in addition, attenuated the activation of NTS neurons, in terms of c-fos and NR1 expressions, and reduced the anxiety behaviors of adult rats subjected to hypobaric hypoxic challenge. Reduction of c-fos immunoreactive neurons was found concentrated in the chemoreceptor, baroreceptor, and tracheobronchial tree NTS subnuclei that receive corresponding afferents. The protective effect was not found in normal adult animals supplemented with AA or αTC a week before hypobaric hypoxic challenge. In short, prenatal and sustained AA or αTC supplement altered NTS substrate and ameliorated animals' reactions to hypobaric hypoxic insult, suggesting that this may be considered to protect animals from hypoxic insults from young to adult.

Inhibition of spontaneous excitatory transmission induced by codeine is independent on presynaptic K+ channels and novel voltage-dependent Ca2+ channels in the guinea-pig nucleus tractus solitarius

The nucleus tractus solitarius (NTS) constitutes the cough reflex arc and is thought to be one of the main sites of codeine's action. We have previously demonstrated using the guinea-pig brainstem slice that codeine inhibits the solitary tract-evoked excitatory postsynaptic currents (EPSCs) in the second-order NTS neurons through activating the presynaptic K+ channels. For further understanding of modulation of synaptic transmission by the antitussive, the effects of codeine (0.3–3.0 mM) on spontaneous EPSCs (sEPSCs) and miniature EPSCs (mEPSCs) were investigated in the NTS neurons of guinea-pigs. Codeine decreased the frequency and amplitude of sEPSCs. This action of codeine was mimicked by specific μ and κ receptor agonists, and blocked by μ and κ receptor antagonists. An agonist of δ receptors was ineffective on sEPSCs. The inhibitory effect of codeine on sEPSCs persisted under perfusion of a K+ channel blocker, 4-aminopyridine. In the presence of tetrodotoxin or Cd2+ which blocks, respectively, the action potential-induced or voltage-dependent Ca2+ entry-induced transmitter release, codeine still had an inhibitory effect on the frequency of mEPSCs without any considerable effect on their amplitude. The present study demonstrates that codeine depresses spontaneous excitatory synaptic transmission in the NTS neurons via presynaptic μ and κ receptors that do not couple with K+ and Ca2+ channels. These results suggest inhibitory modulation of the local circuit activity of NTS neurons by codeine, resulting in suppression of cough reflex.

Hibernation induces pentobarbital insensitivity in medulla but not cortex

The 13-lined ground squirrel (Ictidomys tridecemlineatus), a hibernating species, is a natural model of physiological adoption to an extreme environment. During torpor, body temperature drops to 0-4 degrees C, and the cortex is electrically silent, yet the brain stem continues to regulate cardiorespiratory function. The mechanisms underlying selective inhibition in the brain during torpor are not known. To test whether altered GABAergic function is involved in regional and seasonal differences in neuronal activity, cortical and medullary slices from summer-active (SA) and interbout aroused (IBA) squirrels were placed in a standard in vitro recording chamber. Silicon multichannel electrodes were placed in cortex, ventral respiratory column (VRC), and nucleus tractus solitarius (NTS) to record spontaneous neuronal activity. In slices from IBA squirrels, bath-applied pentobarbital sodium (300 microM) nearly abolished cortical neuronal activity, but VRC and NTS neuronal activity was unaltered. In contrast, pentobarbital sodium (300 microM) nearly abolished all spontaneous cortical, VRC, and NTS neuronal activity in slices from SA squirrels. Muscimol (20 microM; GABA(A) receptor agonist) abolished all neuronal activity in cortical and medullary slices from both IBA and SA squirrels, thereby demonstrating the presence of functional GABA(A) receptors. Pretreatment of cortical slices from IBA squirrels with bicuculline (100 microM; GABA(A) receptor antagonist) blocked pentobarbital-dependent inhibition of spontaneous neuronal activity. We hypothesize that GABA(A) receptors undergo a seasonal modification in subunit composition, such that cardiorespiratory neurons are uniquely unaffected by surges of an endogenous positive allosteric modulator.

Functional interactions between the nucleus tractus solitarius (NTS) and nucleus accumbens shell in modulating memory for arousing experiences

The shell division of the nucleus accumbens receives noradrenergic input from neurons in the nucleus of the solitary tract (NTS) that transmit information regarding fluctuations in peripheral hormonal and autonomic activity. Accumbens shell neurons also receive converging inputs from limbic areas such as the hippocampus and amygdala that process newly acquired information. However, few studies have explored whether peripheral information regarding changes in emotional arousal contributes to memory processing in the accumbens. The beneficial effects on memory produced by emotional arousal and the corresponding activation of NTS neurons may be mediated through influences on neuronal activity in the accumbens shell during memory encoding. To explore this putative relationship, Experiment 1 examined interactions between the NTS and the accumbens shell in modulating memory for responses acquired after footshock training in a water-motivated inhibitory avoidance task. Memory for the noxious shock was significantly improved by posttraining excitation of noradrenergic NTS neurons. The enhanced retention produced by activating NTS neurons was attenuated by suppressing neuronal activity in the accumbens shell with bupivacaine (0.25%/0.5 μl). Experiment 2 examined the direct involvement of accumbens shell noradrenergic activation in the modulation of memory for psychologically arousing events such as a reduction in perceived reward value. Noradrenergic activation of the accumbens shell with phenylephrine (1.0 μg/0.5 μl) produced an enhancement in memory for the frustrating experience relative to control injections as evidenced by runway performance on an extended seven-day retention test. These findings demonstrate a functional relationship between NTS neurons and the accumbens shell in modulating memory following physiological arousal and identifies a role of norepinephrine in modulating synaptic activity in the accumbens shell to facilitate this process.

17β-Estradiol inhibits excitatory amino acid-induced activity of neurons of the nucleus tractus solitarius

The effects of 17β-estradiol (17βE2) on spontaneous and excitatory amino acid (EAA) induced nucleus tractus solitarius (NTS) neuronal activity were investigated by electrophysiological and immunohistochemical experiments in ovariectomized female Sprague–Dawley rats. Out of 62 NTS neurons tested, 42 were inhibited (68%) following iontophoretic application of 17βE2 in a current-dependent manner. The averaged firing rate decreased from 3.06±0.40 to 0.78±0.17 Hz. The inhibitory responses were rapid in onset (within 1 min) and variable in duration (2–4 min). The inhibitory effects of 17βE2 were blocked by simultaneously applied 17βE2 antagonist ICI182,780, but not by GABA antagonist, bicuculline and phaclofen. l-Glutamate, AMPA or NMDA enhanced the activity of 71, 73 or 69% of NTS cells tested, respectively. The excitatory effects of EAA were significantly inhibited in the presence of 17βE2. Fluorescent immunohistochemistry revealed that all subnuclei of the NTS contained high levels of estrogen receptors (ERs) immunoreactivity. These results suggest that 17βE2 inhibits spontaneous and EAA-induced NTS neuronal activity through 17βE2 activation of ERs.

Gastric effects of galanin and its interaction with leptin on brainstem neuronal activity.

Galanin is a 29-amino acid peptide that is widely distributed throughout the central nervous system, peripheral nervous system, and gastrointestinal and genitourinary tracts. Leptin is a hormone secreted from adipose tissue and the gut and other tissues. In this study, using an in vitro neonatal rat preparation, we investigated the gastric effects of galanin and its interaction with leptin on nucleus tractus solitarius (NTS) neurons receiving gastric vagal inputs. We showed that peripheral gastric galanin (300 nM) produced a mean inhibition response of 53.2 +/- 2.1% compared with the control level of 100% (P < 0.01) in 27 of 58 neurons tested. A concentration-dependent effect of galanin on NTS neuronal activity was observed. The galanin receptor antagonist [galanin-(1-12)-Pro3-(Ala-Leu)2-Ala amide], or M40, significantly reversed the galanin-induced inhibition effect (P < 0.01). In contrast, we showed that the peripheral gastric effect of leptin (10 nM) produced a mean activation response of 167.4 +/- 8.2% compared with the control level. The NTS neurons that we recorded could respond to both galanin and leptin or respond to only one of them. Subsequently, we evaluated gastric interactions between galanin and leptin on NTS unitary activity when galanin (100 nM) and leptin (10 nM) were applied together in the gastric compartment. We observed that the effect of leptin when applied alone (168.8 +/- 7.7%) was reduced to 146.2 +/- 4.7% after coapplication of both compounds (P < 0.05 compared with leptin alone; P < 0.01 compared with galanin alone, 55.1 +/- 3.2%). Our data suggest that galanin modulates the leptin signals, which regulate the ingestive process in neonates.