Region Of Nucleus Tractus Solitarius Research Articles

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 γ-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.

CO2 dialysis in nucleus tractus solitarius region of rat increases ventilation in sleep and wakefulness.

To evaluate the function of widely distributed central chemoreceptors during sleep and wakefulness in the rat, we focally stimulate single chemoreceptor sites during naturally occurring sleep-wake cycles by microdialysis of artificial cerebrospinal fluid equilibrated with 25% CO2. In retrotrapezoid nucleus, this increased ventilation (tidal volume) by 24% only in wakefulness (Li A, Randall M, and Nattie E. J Appl Physiol 87: 910-919, 1999). In caudal medullary raphé, it increased ventilation (frequency) by 15-20% only in sleep (Nattie EE and Li A. J Appl Physiol 90: 1247-1257, 2001). Here, in nucleus tractus solitarius (NTS), focal acidification significantly increased ventilation by 11% in sleep and 7% in wakefulness rostrally (n = 5) and by 16% in sleep and 28% in wakefulness caudally (n = 5). The sleep-wake cycle was unaltered. Dialysis with 5% CO2 had no effect. Dialysis with 50% CO2 caudally did not further stimulate ventilation but did disrupt sleep. Central chemoreceptors in the NTS affect breathing in both sleep and wakefulness. The threshold for arousal in caudal NTS is greater than that for the stimulation of breathing.

Differential CO(2)-induced c-fos gene expression in the nucleus tractus solitarii of inbred mouse strains.

Genetic determinants confer variation between inbred mouse strains with respect to the magnitude and pattern of ventilation during hypercapnic challenge. Specifically, inheritance patterns derived from low-responsive C3H/HeJ (C3) and high-responsive C57BL/6J (B6) mouse strains suggest that differential hypercapnic ventilatory sensitivity (HCVS) is controlled by two independent genes. The present study also tests whether differential neuronal activity in respiratory control regions of the brain is positively associated with strain variation in HCVS. With the use of whole body plethysmography, ventilation was assessed in C3 and B6 strains at baseline and during 30 min of hypercapnia (inspired CO(2) fraction = 0.15, inspired O(2) fraction = 0.21 in N(2)). Subsequently, in situ hybridization histochemistry was performed to determine changes in c-fos gene expression in the commissural subnucleus of the nucleus tractus solitarius (NTS). During hypercapnia, breathing frequency and tidal volume were significantly (P < 0.01) different between strains: C3 mice showed a slow, deep-breathing pattern relative to a rapid, shallow phenotype of B6 mice. CO(2)-induced increase in c-fos gene expression was significantly (P < 0.01) greater in NTS regions of B6 compared with C3 mice. In this genetic model of differential HCVS, the results suggest that a genomic basis for varied hypercapnic chemoreception or transduction confers greater afferent neuronal activity in the caudal NTS for high-responsive B6 mice compared with low-responsive C3 mice.

NO control of respiration

Recent studies suggest that haemoglobin's original role was to promote nitric oxide-mediated reactions key to respiration, and that the ability to deliver oxygen evolved later. In the September edition of Nature (413, 171–174), Lipton et al. have shown that S-nitrosothiols (SNOs) act directly within the brainstem to control breathing. The respiratory cycle begins when haemoglobin is oxygenated in the lungs and cysteine residues within the molecule are modified by NO to SNO. Haemoglobin will then carry the disguised duo throughout the body, ultimately releasing oxygen in the brain by undergoing a structural change that depends on transnitrosylation of its NO group to a glutathione molecule. This reaction triggers γ-glutamyltranspeptidase in neuronal tissues to cleave S-nitrosoglutathione into S-nitrosocysteinyl glycine and S-nitrosocysteine, both stereoactive molecules that modify specific target proteins in the nucleus tractus solitarius region of the brainstem and regulate the central neuronal circuitry that controls breathing. SS

Reliability of monosynaptic sensory transmission in brain stem neurons in vitro.

The timing of events within the nervous system is a critical feature of signal processing and integration. In neurotransmission, the synaptic latency, the time between stimulus delivery and appearance of the synaptic event, is generally thought to be directly related to the complexity of that pathway. In horizontal brain stem slices, we examined synaptic latency and its shock-to-shock variability (synaptic jitter) in medial nucleus tractus solitarius (NTS) neurons in response to solitary tract (ST) electrical activation. Using a visualized patch recording approach, we activated ST 1-3 mm from the recorded neuron with short trains (50-200 Hz) and measured synaptic currents under voltage clamp. Latencies ranged from 1.5 to 8.6 ms, and jitter values (SD of intraneuronal latency) ranged from 26 to 764 micros (n = 49). Surprisingly, frequency of synaptic failure was not correlated with either latency or jitter (P > 0.147; n = 49). Despite conventional expectations, no clear divisions in latency were found from the earliest arriving excitatory postsynaptic currents (EPSCs) to late pharmacologically polysynaptic responses. Shortest latency EPSCs (<3 ms) were mediated by non-N-methyl-D-aspartate (non-NMDA) glutamate receptors. Longer latency responses were a mix of excitatory and inhibitory currents including non-NMDA EPSCs and GABAa receptor-mediated currents (IPSC). All synaptic responses exhibited prominent frequency-dependent depression. In a subset of neurons, we labeled sensory boutons by the anterograde fluorescent tracer, DiA, from aortic nerve baroreceptors and then recorded from anatomically identified second-order neurons. In identified second-order NTS neurons, ST activation evoked EPSCs with short to moderate latency (1.9-4.8 ms) but uniformly minimal jitter (31 to 61 micros) that were mediated by non-NMDA receptors but had failure rates as high as 39%. These monosynaptic EPSCs in identified second-order neurons were significantly different in latency and jitter than GABAergic IPSCs (latency, 2.95 +/- 0.71 vs. 5.56 +/- 0.74 ms, mean +/- SE, P = 0.027; jitter, 42.3 +/- 6.5 vs. 416.3 +/- 94.4 micros, P = 0.013, n = 4, 6, respectively), but failure rates were similar (27.8 +/- 9.0 vs. 9.7 +/- 4.4%, P = 0.08, respectively). Such results suggest that jitter and not absolute latency or failure rate is the most reliable discriminator of mono- versus polysynaptic pathways. The results suggest that brain stem sensory pathways may differ in their principles of integration compared with cortical models and that this importantly impacts synaptic performance. The unique performance properties of the sensory-NTS pathway may reflect stronger axosomatic synaptic processing in brain stem compared with dendritically weighted models typical in cortical structures and thus may reflect very different strategies of spatio-temporal integration in this NTS region and for autonomic regulation.

Morphological Investigation of Nucleus Tractus Solitarius Somatotopy.

Taste is one of the most important senses that allows for human survival. Recent electrophysiological investigations have revealed some information about the somatotopy of the rat nucleus tractus solitarius (NTS). The aim of this study was to assess rat NTS somatotopy using morphological techniques. Each animal was anesthetized and a retrograde axonal tracer, horseradish peroxidase, was injected intra- and submucosally into the anterior portion or the middle and posterior portions of the left half of rat tongue. After 24-48 hr, each rat was anesthetized again and perfusion-fixed. The brainstem was removed and cut in 50 μm-thick sections using a vibrotome. The sections were subjected to a tetramethylbenzidine-peroxidase reaction, and were then assessed under light-microscopy. The horseradish peroxidase that was deposited in the anterior third of the tongue labeled neuron cell bodies in the rostrofrontolateral region of the ipsilateral NTS, while that deposited in the middle and posterior thirds of the tongue labeled neuron cell bodies in the caudoposteromedial region of the ipsilateral NTS. There was no cross-projection. The morphological findings were in accord with the electrophysiological results in the literature. Retrograde labeling revealed a distinct pattern of neural projection that corresponded to the anterior portion, and the middle and posterior portions of the tongue.

Distribution and classification of respiratory neurones in the nucleus tractus solitarius of the rat medulla and their responses to central and periphereal stimuli

In the cat the dorsal respiratory group comprising the nucleus tractus solitarius (NTS) is regarded as an important respiratory centre. However the relevance of this nucleus in the control of respiration in the rat is still unclear. The present work aimed to identify and localize the various neurons involved in respiration in the nucleus tractus solitarius region of the rat medulla. Further, we intended to study the responses of these identified respiratory neurons to central and peripheral stimulation, in particular: 1)effect of stimulation of midbrain periaqueductal gray (PAG) matter and, 2)effect of stimulation of the vagus (vagal feedback). Nembutal-anaesthetised (80 mg/Kg), spontaneously-breathing rats (n = 60, 300–400 g) were used for the study. Neurons firing in relation to diaphragm electromyogram (dEMG) were located between 0.6–1.5 mm lateral to the midline, extending 0.5 mm caudal to 1.8 mm rostral to the posterior end of the area postrema (AP). Four different populations of respiratory-related neurons were recorded. They have been classified (with reference to dEMG) as: expiratory-inspiratory phase-spanning cells, early inspiratory neurons (eI-cells), cells that fire only in phase with diaphragm (I-all cells), and expiratory cells. The phase-spanning and eI-cell populations were located both in the ventrolateral and ventromedial NTS rostral to the area postrema, while the other cells were found predominantly in the more lateral regions. eI-cells were observed to start firing up to 20 ms prior to dEMG and ceased firing about one third of the way through diaphragm activity. They also showed a decrementing discharge pattern. Excitatory amino acid (DLH, 15–60 nl of 0.2 M) stimulation of PAG along its dorso-ventral axis produced dose-dependent changes in the cardiorespiratory parameters: increased respiratory frequency, decreased inspiratory and expiratory durations, increased blood pressure (BP) and heart rate. PAG stimulation elicited increases in activity in both phase-spanning and eI-neuronal populations in the NTS. The PAG sites evoking an increase in the phase-spanning cell activity were located more laterally than the PAG sites that led to increased eI-cell activity. The increase in the responses of the phase-spanning neurons was not accompanied by any changes in the respiratory frequency, whereas the PAG stimulation giving rise to an increase in eI-neuronal activity also elicited simultaneous increases in dEMG and the respiratory frequency. The central end of the cut vagus was electrically stimulated to activate pulmonary stretch-receptor afferents. Vagal stimulation caused depression of inspiratory cell activity along with bradycardia and a fall in BP. Both eI-neurons and I-all cells were inhibited. These cells exhibited a 'vagal escape' phenomenon ie, resuming activity whilst vagal stimulation continued. Phase-spanning cells and expiratory cells are yet to be tested for responses to vagal stimulation. The behaviours of these respiratory-related cells of the NTS, their temporal relationships with the dEMG, firing patterns, their responses to central and peripheral stimuli are discussed and a possible role for each in the brainstem control of respiration is proposed. Similar studies have also been carried out to elucidate the roles of ventrolateral column (VLM)/ventral respiratory group (VRG) neurons in the control of respiration in the rat. The findings of these studies are presented in an accompanying abstract.

Involvement of apamin-sensitive SK channels in spike frequency adaptation of neurons in nucleus tractus solitarii of the rat.

We delineated the role of Ca(2+)-activated K(+) channels in the phenomenon of spike frequency adaptation (SFA) exhibited by neurons in the caudal region of nucleus tractus solitarius (cNTS) using intracellular recording coupled with the current-clamp technique in rat brain slices. Intracellular injection of a constant depolarizing current evoked a train of action potentials whose discharge frequency declined rapidly to a lower steady-state level of irregular discharges. This manifested phenomenon of SFA was found to be related to extracellular Ca(2+). Low Ca(2+) (0.25 mM) or Cd(2+) (0.5 mM) in the perfusing medium resulted in a significant increase in the adaptation time constant (tau(adap)) and an appreciable reduction in the percentage adaptation of spike frequency (F(adap)). In addition, the evoked discharges were converted from an irregular to a regular pattern, accompanied by a profound increase in mean firing rate. Intriguingly, similar alterations in tau(adap), F(adap), discharge pattern and discharge rate were elicited by apamin (1 microM), a selective blocker for small-conductance Ca(2+)-activated K(+) (SK) channels. On the other hand, charybdotoxin (0.1 microM), a selective blocker for large-conductance Ca(2+)-activated K(+) channels, was ineffective. Our results suggest that SK channels of cNTS neurons may subserve the generation of both SFA and irregular discharge patterns displayed by action potentials evoked with a prolonged depolarizing current.

Involvement of Apamin-Sensitive SK Channels in Spike Frequency Adaptation of Neurons in Nucleus tractus solitarii of the Rat

We delineated the role of Ca²⁺-activated K⁺ channels in the phenomenon of spike frequency adaptation (SFA) exhibited by neurons in the caudal region of nucleus tractus solitarius (cNTS) using intracellular recording coupled with the current-clamp technique in rat brain slices. Intracellular injection of a constant depolarizing current evoked a train of action potentials whose discharge frequency declined rapidly to a lower steady-state level of irregular discharges. This manifested phenomenon of SFA was found to be related to extracellular Ca²⁺. Low Ca²⁺ (0.25 mM) or Cd²⁺ (0.5 mM) in the perfusing medium resulted in a significant increase in the adaptation time constant (τ_adap) and an appreciable reduction in the percentage adaptation of spike frequency (F_adap). In addition, the evoked discharges were converted from an irregular to a regular pattern, accompanied by a profound increase in mean firing rate. Intriguingly, similar alterations in τ_adap, F_adap, discharge pattern and discharge rate were elicited by apamin (1 μM), a selective blocker for small-conductance Ca²⁺-activated K⁺ (SK) channels. On the other hand, charybdotoxin (0.1 μM), a selective blocker for large-conductance Ca²⁺-activated K⁺ channels, was ineffective. Our results suggest that SK channels of cNTS neurons may subserve the generation of both SFA and irregular discharge patterns displayed by action potentials evoked with a prolonged depolarizing current.

Dorsal medullary 5-HT3 receptors and sympathetic premotor neurones in the rat.

1. Our aim was to determine whether the cardiovascular neurones in the rostro-ventrolateral medulla (CV-RVLM neurones) were involved in the sympathoexcitation induced by stimulation of 5-HT3 receptors in the region of the nucleus tractus solitarii (NTS). Experiments were performed in pentobarbitone-anaesthetized rats, artificially ventilated and paralysed with pancuronium bromide. 2. Using extracellular recordings, different types of RVLM neurones were characterized: cardiovascular (CV), ventilation-related and baroreflex-insensitive (unidentified) neurones. The CV-RVLM cells were further subdivided into three populations according to their axonal conduction velocities: A (1.2 +/- 0.1 m s-1), B (2.5 +/- 0.2 m s-1) and C (6.8 +/- 1.1 m s-1). 3. Only the CV-RVLM neurones of the A and B categories were partially inhibited (-30 %) by a hypotensive dose (2.5 microg kg-1 i.v.) of clonidine. 4. Microinjections into the region of the commissural NTS of 1-(m-chlorophenyl)-biguanide (CPBG, 2 nmol), a selective 5-HT3 receptor agonist, elicited an increase in both lumbar sympathetic nerve discharge (SND) and arterial pressure. In addition, this treatment produced a marked excitation of CV-RVLM neurones of the A and B categories, without affecting those of the C type, as well as ventilation-related and unidentified RVLM cells. 5. The activity of the CV neurones in the caudo-ventrolateral part of the medulla oblongata (CV-CVLM) was not modified by 5-HT3 receptor stimulation in the NTS. 6. Prior intra-NTS microinjections of ondansetron (300 pmol, a selective 5-HT3 receptor antagonist) into the region of the commissural NTS prevented the excitation of A and B CV-RVLM neurones induced by CPBG. 7. Intracarotid administration of saline saturated with CO2 (chemoreceptor activation) elicited both an increase in the SND and an excitation of the clonidine-insensitive CV-RVLM neurones of the C type, without affecting A and B neurones. 8. In conclusion, the sympathoexcitation elicited following 5-HT3 receptor stimulation in the region of the commissural NTS of pentobarbitone-anaesthetized rats seems to result from the excitation of two different pools of clonidine-sensitive CV-RVLM neurones. These neurones are apparently not involved in the sympathetic component of the chemoreceptor reflex.

Exposure to Acute Stress Induces Brain Interleukin-1β Protein in the Rat

Peripheral immune stimulation such as that provided by lipopolysaccharide (LPS) has been reported to increase brain levels of IL-1beta mRNA, immunoreactivity, and bioactivity. Stressors produce many of the same neural and endocrine responses as those that follow LPS, but the impact of stressors on brain interleukin-1beta (IL-1beta) has not been systematically explored. An ELISA designed to detect IL-1beta was used to measure levels of IL-1beta protein in rat brain. Brain IL-1beta was explored after exposure to inescapable shock (IS; 100 1.6 mA tail shocks for 5 sec each) and LPS (1 mg/kg) as a positive control. Rats were killed either immediately or 2, 7, 24, or 48 hr after IS. Brains were dissected into hypothalamus, hippocampus, cerebellum, posterior cortex, and nucleus tractus solitarius regions. LPS produced widespread increases in brain IL-1beta, but IS did not. Adrenal glucocorticoids are known to suppress IL-1beta production in both the periphery and brain. Thus, it was possible that the stressor did provide stimulus input to the brain IL-1beta system(s), but that the production of IL-1beta protein was suppressed by the rapid and prolonged high levels of glucocorticoids produced by IS. To test this possibility rats were adrenalectomized or given sham surgery, with half of the adrenalectomized rats receiving corticosterone replacement to maintain basal corticosterone levels. IS produced large increases in brain IL-1beta protein in the adrenalectomized subjects 2 hr after stress, whether basal corticosterone levels had been maintained. Thus elimination of the stress-induced rise in corticosterone unmasked a robust and widespread increase in brain IL-1beta.

Non-NMDA receptors transmit cardiopulmonary C fibre input in nucleus tractus solitarii in rats.

1. We sought first to determine whether neurones in caudomedial aspects of commissural nucleus tractus solitarii (NTS) received input from cardiopulmonary C fibre endings supplied by the pulmonary versus systemic circulation. We then examined the role of N-methyl-D-aspartate (NMDA) and non-NMDA receptors in transmitting cardiopulmonary C fibre input to such NTS neurones. 2. Extracellular NTS unit activity, phrenic nerve activity and arterial blood pressure were recorded in urethane-anaesthetized rats. Unit responses to right atrial and left ventricular phenylbiguanide injections were compared in rats with arterial baroreceptors, carotid chemo-receptors and subdiaphragmatic vagal inputs eliminated. Right atrial phenylbiguanide injections produced greater peak responses (27 +/- 11 impulses s-1) than did left ventricular injections (11 +/- 3 impulses s-1) (n = 9). 3. The non-NMDA receptor agonist quisqualic acid (QUIS) and NMDA were ionophoresed onto NTS neurones that were synaptically activated by right atrial phenylbiguanide injection. Responses were compared before and during ionophoresis of the non-NMDA receptor antagonist 2,3-dihydroxy-6-nitro-7-sulphamoylbenzo(f)quinoxaline (NBQX), the NMDA receptor antagonist DL-2-amino-5-phosphovaleric acid (AP5), or the broad spectrum antagonist kynurenic acid (KYN). 4. NBQX, which blocked QUIS-but spared NMDA-evoked responses, significantly attenuated synaptic activation by 65% (n = 9). AP5, which blocked NMDA- but spared QUIS-evoked responses, did not significantly diminish synaptic activation (11%; n = 7). KYN, which blocked QUIS- and NMDA-evoked responses, decreased synaptic activation by 70% (n = 9). 5. The results suggest that input from cardiopulmonary C fibre endings, primarily supplied by the pulmonary circulation, is transmitted to this commissural NTS region largely via non-NMDA receptors.

The role of raphe and tractus solitarius neuronal structures in the modulation of respiratory pattern in rabbits.

We studied the effects of MNR stimulation on phrenic (Phr) electroneurogram and external intercostal muscles (EI) electromiogram in spontaneously breathing rabbits. Additionally, experiments were performed before and after lignocaine blockade of nucleus tractus solitarii (NTS) to determine whether the information from MNR is transmitted via NTS neurones. The completeness of the blockade of NTS region was checked by studying the Hering-Breuer reflex. MNR was stimulated at the level 2-7 mm rostral to the obex. Stimulation at the rostral part of this region produced inhibition of phasic inspiratory activity, whereas stimulation in the caudal part elicited tonic activity throughout the respiratory cycle. These effects were more pronounced on EI than Phr. Responses to MNR stimulation were attenuated after lignocaine blockade, suggesting that the neurons located in NTS take part in the transmission of the modulatory information from the MNR to respiratory motoneurones.

Phenotypic characteristics of expressed tyrosine hydroxylase protein in the adult rat nucleus tractus solitarius: Plasticity revealed by RU24722 treatment

The phenotypic characteristics of expressed tyrosine hydroxylase protein have been precisely analysed in the rat nucleus tractus solitarius, which contains the majority of A2 noradrenergic and C2 adrenergic neurons of the medulla oblongata. This study was based upon quantitative analysis of immunohistochemical and immunoradioautographic staining of tyrosine hydroxylase protein in serial coronal sections. In control rats, there were few tyrosine hydroxylase-expressing cell bodies which express less than 2% of the immunoradiolabeled tyrosine hydroxylase protein measured in the structure. These cell bodies were scattered throughout an extensive immunopositive neuropile, which precisely delimited the topological space of the nucleus tractus solitarius quantitatively reconstructed using a polar coordinate system. The quantification of tyrosine hydroxylase tissue concentration from immunoradioautograms allowed us to subdivide the structure into two distinct regions. The posterior region of the nucleus tractus solitarius, which mainly corresponds to the A2 cell group, contains a relatively high tissue concentration of tyrosine hydroxylase protein (18.56 ± 0.154 units per mg of tissue). The anterior region, which mainly corresponds to the C2 cell group, exhibits a relatively low concentration (12.09 ± 0.81) of this protein. Three days after an intraperitoneal injection of RU24722, there was a strong increase (90 ± 17%) in tyrosine hydroxylase protein content only in the anterior region of the nucleus tractus solitarius. This increase was associated with a dramatic elevation (142 ± 20%) in the number of tyrosine hydroxylase-expressing cell bodies. The additional cell bodies were mainly located inside the initial perikarya-containing area. Furthermore, in this region a large and preferential lateral extension in the mean topological space of the nucleus tractus solitarius as defined by tyrosine hydroxylase radiolabeling was also quantified. These results suggested a particular plasticity in tyrosine hydroxylase protein expression in the anterior adrenergic region of the rat nucleus tractus solitarius.

Effects of plasma hypernatremia on nucleus tractus solitarius neurons

Experiments were done in chloralose-anesthetized, paralyzed, and artificially ventilated rats to investigate the effect of plasma hypernatremia and baroreceptor activation on the excitability of nucleus tractus solitarius (NTS) neurons. Extracellular, single unit recordings were made from 67 histologically verified neurons in the region of NTS. The firing frequency of 29 (43%) NTS neurons was increased by the intracarotid infusion of hypertonic saline (0.5 M). The acute activation of baroreceptors after the intravenous infusion of phenylephrine resulted in the excitation of seven (31.8%) or the inhibition of five (22.7%) of the units responsive to plasma hypernatremia. Thirteen of the hypertonic saline-responsive neurons were recorded in animals in which the organum vasculosum lamina terminalis and subfornical organ were lesioned and/or the left vagus nerve was cut. These data suggest that within NTS there exists a pool of neurons that alter their discharge rate in response to changes in plasma sodium levels and that these neurons may also function in cardiovascular regulation.

Target site of inhibition mediated by midbrain periaqueductal gray matter of baroreflex vagal bradycardia.

1. Both electrical and chemical stimulation of the midbrain periaqueductal gray matter (PAG) inhibit baroreflex vagal bradycardia (BVB). The present study was designed to determine the target site of this inhibition about which little is known. Electrical stimulation of the PAG, in particular of its dorsal portion, markedly suppressed BVB provoked by electrical stimulation of the aortic depressor nerve (ADN; percentage of inhibition = 91.0 +/- 9.7%, mean +/- SD; n = 64). To identify the target site of the inhibition, several types of experiments were conducted in rats under chloralose-urethan anesthesia. 2. The inhibition was exclusively of central origin because inhibition of BVB by stimulation of the PAG was unchanged after transection of the spinal cord at the C1 level. According to Wall's method, we examined whether PAG stimulation affects BVB presynaptically by modulating the excitability of ADN terminals in the nucleus tractus solitarius (NTS). However, excitability changes of ADN terminals by the PAG stimulation were not demonstrated. 3. Vagal bradycardia evoked by microinjection of glutamate into the nucleus ambiguus (NA) region was markedly suppressed by the PAG (percentage of inhibition = 85.9 +/- 9.1%; n = 9), an indication that vagal cardiac preganglionic neurons at this site were subject to the inhibitory action of the PAG. Basal vagal tone due to ongoing preganglionic neuronal activity was also subject to inhibitory control by the PAG because basal heart rate was increased by stimulation of the PAG after either C1 transection or NTS lesion. 4. We found that PAG stimulation suppressed ADN-induced field potentials in the NA region (37.7 +/- 13.8% relative to the control; n = 9) but only slightly in the NTS region (95.8 +/- 15.2%; n = 16). In addition, unitary recordings revealed that ADN-evoked unitary responses of neurons in the NA region were suppressed by PAG stimulation, whereas NTS baroreceptor neurons, either ADN responsive or nonresponsive, were scarcely inhibited by PAG stimulation. 5. These findings suggest that the PAG inhibited BVB mainly at the vagal preganglionic cell level and not at the NTS interneuron level. The conclusion is in harmony with our previous reports that the target site of hypothalamic inhibition of BVB in rats is also the preganglionic neurons and that hypothalamic inhibition of BVB is mediated predominantly by the PAG.

Endothelin concentrations in respiration-related structures of the medulla during the perinatal period of the rat

Endothelin-like immunoreactivity (ET-LI) was quantified in the developing (foetal and postnatal) rat brain stem and cerebellum using radioimmunoassay. The brain stem structures chosen for this study were (a) dorsal medullary region (DMR) including the region of nucleus tractus solitarius where the peripheral chemoreceptor afferents are known to terminate, (b) ventral medullary region (VMR) where the central chemoreceptors are thought to be located and (c) cerebellum (CER), as a control area. Compared to the prenatal period, significantly elevated concentrations of ET-LI were detected in the early postnatal period and thereafter the concentrations decreased: DMR and VMR: in comparison to the prenatal concentrations, a two-fold increase was found on the day of birth which further increased significantly ( P < 0.001) on postnatal day 1 only in the region of DMR; CER: low concentrations of ET-LI were found in the early postnatal period which were not significantly different from the prenatal values. No ET-LI could be detected in any of the three regions in the adult rats. The results are discussed in view of the hypothesis that (1) endothelin appears to play an important role in the perinatal period and (2) it is involved in the chemoreceptor pathway.

CNS innervation of airway-related parasympathetic preganglionic neurons: a transneuronal labeling study using pseudorabies virus

The CNS cell groups that innervate the tracheal parasympathetic preganglionic neurons were identified by the viral retrograde transneuronal labeling method. Pseudorabies virus (PRV) was injected into the tracheal wall of C8 spinal rats and after 4 days survival, brain tissue sections from these animals were processed for immunohistochemical detection of PRV. Retrogradely labeled parasympathetic preganglionic neurons were seen in three sites in the medulla: the compact portion of the nucleus ambiguus, the area ventral to the nucleus ambiguus, and the rostralmost portion of the medial nucleus tractus solitarius (NTS); this labeling pattern correlated well with the retrograde cell body labeling seen following cholera toxin ß-subunit injections in the tracheal wall. PRV transneuronally labeled neurons were found throughout the CNS with the most abundant labeling concentrated in the ventral medulla oblongata. Labeled neurons were identified along the ventral medullary surface, and in nearby areas including the parapyramidal, retrotrapezoid, gigantocellular and lateral paragigantocellular reticular nuclei as well as the caudal raphe nuclei (raphe pallidus, obscurus, and magnus). Serotonin (5-HT) neurons of the caudal raphe complex (B1–B3 cell groups) and ventromedial medulla were labeled as well as a few C1 adrenergic neurons. The A5 cell group was the major noradrenergic area labeled although a small number of locus coeruleus neurons were also labeled. Several NTS regions contained labeled cells including the commissural, intermediate, medial, central, ventral, and ventrolateral subnuclei. PRV infected neurons were present in the Kölliker-Fuse and Barrington's nuclei. In the rostral mesencephalon, the precommissural nucleus of the dorsal periventricular gray matter was labeled. Labeling was present in the dorsal, lateral and paraventricular hypothalamic nuclei. In summary, the airway parasympathetic preganglionic neurons are innervated predominantly by a network of lower brainstem neurons that lie in the same regions known to be involved in respiratory and cardiovascular regulation. These findings are discussed in relationship to some of the potential CNS mechanisms that may be operative in airway disorders as well as potentially involved in certain fatal respiratory conditions such as Ondine's curse and sudden infant death syndrome (SIDS).

An atlas of the rat subpostremal nucleus tractus solitarius

The nucleus tractus solitarius (NTS) in the dorsal medulla is the principal visceral sensory relay nucleus in the brain. In the rat, numerous lines of evidence indicate that the caudal NTS at the level of the area postrema serves as a major integrating site for coordinating cardiorespiratory reflexes and viscerobehavioral responses. This region of the caudal NTS not only exhibits high densities of binding sites for an impressive array of transmitters and modulators but microinjections of many of these same neuroactive substances into the rat subpostremal NTS elicit pronounced cardiorespiratory and visceral response patterns. This report provides an abbreviated atlas of the rat subpostremal NTS consisting of a series of transverse, sagittal, and horizontal plates. Photomicrographs, together with their corresponding schematic drawings, are provided for the serial sections generated from each reference plane.

Contribution of catecholaminergic neurons of the dorsomedial and ventrolateral medulla oblongata to the hypotensive effect of clonidine in spontaneously hypertensive rats: in vivo voltammetric studies.

Our previous electrochemical studies in the normotensive Wistar-Kyoto (WKY) rat showed a positive correlation between the hypotensive effect of low doses of clonidine (2-10 micrograms/kg intravenously) and inhibition of the activity of catecholaminergic neurons within the brainstem, and that this action was mediated by imidazoline-preferring receptors. In the present study the possibility of a relationship between the centrally mediated hypotensive effect of clonidine and the metabolic activity of catecholaminergic neurons of the ventrolateral and dorsomedial medulla oblongata was investigated in pentobarbital-anaesthetized spontaneously hypertensive rats (SHR). Neuronal metabolic activity was monitored by in vivo electrochemistry in the nucleus reticularis lateralis region of the ventrolateral medulla and in the nucleus tractus solitarii region of the dorsomedial medulla oblongata. Hypotensive doses of intravenously administered clonidine (10 micrograms/kg) failed to inhibit the neuronal metabolic activity of the nucleus reticularis lateralis region; fivefold higher doses were required to inhibit these neurons. The low dose of clonidine (10 micrograms/kg) decreased the metabolic activity of the nucleus tractus solitarii region. Nevertheless, this effect did not appear to be correlated with a reduction in blood pressure and it was not attenuated by a prior central injection of idazoxan (5 nmol/kg intracisternally), which almost completely prevented the hypotensive and the neuronal inhibitory effects of clonidine in the nucleus reticularis lateralis region of the normotensive rat. Both the site and the mechanism of the central hypotensive action of clonidine in the SHR appear to be different from those in normotensive control WKY rats.