The projections to the medulla of neurons innervating the carotid sinus in the dog

The localization and brain stem projections of neurons innervating the carotid sinus of the dog were studied by horseradish peroxidase histochemistry following microinjection of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) under the adventitia of the carotid sinus. Within the brain stem, labeled afferent fibers and presumptive terminals were found bilaterally in the caudal nucleus tractus solitarius (nTS), the area postrema (AP), and the lateral tegmental field (LTF), reaching the area of the nucleus ambiguus (nA). Sparse labeling was also seen in the ipsilateral spinal trigeminal nucleus (SpV) and lateral cuneatus nucleus (LCn). These findings suggest the existence of multiple pathways by which peripheral baroreceptor inputs may influence central cardiovascular-related neurons. In addition to classically defined relay in the nTS, carotid sinus afferents may also interact more directly with these neurons in other brain stem regions.     

The central distribution of the cervical vagus nerve and gastric afferent and efferent projections in the rat

The medullary distribution of afferent fibers and cells of origin of the cervical vagal trunk and of the vagal innervation of the stomach have been studied using the anterograde and retrograde transport of horseradish peroxidase (HRP). Injections of HRP were made into the cervical vagus nerve, the stomach wall, the proximal small intestine, or the peritoneal cavity. Two to four days following the injections, the rats were perfused and the medullae oblongatae and nodose ganglia were processed using the tetramethyl benzidine method. Cervical vagus nerve injections of HRP resulted in heavy anterograde labeling in the ipsilateral nucleus of the tractus solitarius (NTS) and the commissural nucleus. Lighter labeling was seen in these regions on the contralateral side, but did not extend as far rostrally in the NTS. Labeling was also seen in the area postrema. Retrogade labeling of somata was present in the ipsilateral side in the nodose ganglion, throughout the whole extent of the dorsal motor nucleus of the vagus, much of the nucleus ambiguus and in rostral levels of the cervical spinal cord. After stomach injections, labeling indicative of afferent fibers was observed bilaterally in the dorsomedial and medial portions of the NTS and in the commissural nucleus. Labeled efferent fibres arose from neurons in the dorsal motor nucleus of the vagus, nucleus ambiguus and the cervical spinal cord. Retrogradely labeled somata were found bilaterally, throughout the rostrocaudal length of the dorsal motor nucleus in all cases with stomach injections. In some, but not all cases, labeled somata were seen bilaterally in compact areas within the nucleus ambiguus, particularly rostrally. Control injections of HRP into the intestinal wall and peritoneal cavity indicated that the stomach was the primary source of afferent and efferent labeling in the medulla following subdiaphragmatic injections.     

Mapping of carotid baroreceptor subtype projections to the nucleus tractus solitarius using c-fos immunohistochemistry

This study has combined physiological pressure stimulation of carotid baroreceptors via a vascularly isolated carotid sinus and anodal block of baroreceptor afferent fibers in the carotid sinus nerve to examine the medullary projections of type I vs. type II (large A- vs. small A- and C-fiber afferent) baroreceptors. The control distribution of cells in the nucleus tractus solitarius expressing c-fos in response to physiological activation of carotid baroreceptors in the isolated sinus was compared to that during anodal block of large A-fibers in the carotid sinus nerve. Carotid baroreceptor stimulation primarily activated neurons in the ipsilateral commissural and medial subnuclei of the caudal nucleus tractus solitarius and the dorsolateral, dorsomedial and medial subnuclei in the intermediate and rostral levels of the nucleus tractus solitarius. Elimination of large A-fiber carotid baroreceptor afferents, during similar carotid baroreceptor stimulation resulted in a decrease in the nmber of cells expressing c-fos in the dorsomedial subnucleus of the rostral nucleus tractus solitarius. These data indicate that projections of larger A-fiber (type I) carotid baroreceptors are localized primarily to the rostral dorsomedial subnucleus, while those of smallr A- and C-fiber baroreceptors are more widely distributed to the commissural, medial and dorsal subnuclei of the nucleus tractus solitarius.     

Multimodal responses of taste neurons in the frog nucleus tractus solitarius

The responses of 216 neurons in the nucleus tractus solitarius (NTS) of the American bullfrog were recorded following taste, temperature, and tactile stimulation. Cells were classified on the basis of their responses to 5 taste stimuli: 0.5 M NaCl, 0.0005 M quinine-HCl (QHCl), 0.01 M acetic acid, 0.5 M sucrose, and deionized water (water). Neurons showing excitatory responses to 1, 2, 3, or 4 of the 5 kinds of taste stimuli were named Type I, II, III, or IV, respectively. Cells whose spontaneous rate was inhibited by taste and/or tactile stimulation of the tongue were termed Type V. Type VI neurons were excited by tactile stimulation alone. Of the 216 cells, 115 were excited or inhibited by taste stimuli (Types I-V), with 35 being Type I, 34 Type II, 40 Type III, 2 Type IV and 4 Type V. The remaining 101 cells were responsive only to tactile stimulation (Type VI). Of those 111 cells excited by taste stimulation (Types I-IV), 106 (95%) responded to NaCl, 66 (59%) to acetic acid, 44 (40%) to QHCl, 10 (9%) to water, and 9 (8%) to warming. No cells responded to sucrose. Of the 111 cells of Types I-IV, 76 (68%) were also sensitive to mechanical stimulation of the tongue. There was some differential distribution of these neuron types within the NTS, with more narrowly tuned cells (Type I) being located more dorsally in the nucleus than the more broadly tuned (Type III) neurons. Cells responding exclusively to touch (Type VI) were also more dorsally situated than those responding to two or more taste stimuli (Types II and III).     

The carotid sinus connections: a WGA-HRP study in the cat

The neural connections of the carotid sinus were studied by administration of horseradish peroxidase or a lectin conjugate to the adventitia of the carotid sinus of cats. The carotid sinus afferents project exclusively to the nucleus of the tractus solitarius (NTS). Rostral to the obex the projection is mainly ipsilateral with a strong contralateral component caudal to the obex. The carotid sinus projects to several NTS territories that do not receive afferents from the carotid body chemoreceptors; they are: the dorsolateral, the lateral extension of the commissural, the caudal intermediate, the ventrolateral and the gelatinosus subnuclei. In addition the carotid sinus central representation includes territories occupied also by carotid body terminals: dorsal, interstitial, rostral intermediate, medial and the medial part of commissural subnuclei (see previous paper). Labeled cell bodies were found in the petrosal (216 +/- 37, mean +/- S.E.M.) nodose (825 +/- 434) and superior cervical ganglia (3583 +/- 1227) demonstrating the sympathetic efferent innervation of the carotid sinus and a dual sensory innervation via both the glossopharyngeal and vagus nerves.     

迷走神经参与胃伤害性信息向下丘脑的传递

目的　 研究迷走神经是否参与胃伤害性信息向下丘脑室旁核的传递。 方法　 检测下列条件下c Fos蛋白在孤束核及下丘脑室旁核的表达 :①胃内注入福尔马林引起伤害性刺激 ;②福尔马林刺激结合双侧膈下迷走神经切断术。结果　胃内注入福尔马林引起的伤害性刺激可以诱导c Fos蛋白在孤束核和下丘脑室旁核等脑区的表达 ,但在胸段脊髓的I,V ,VII和X层无明显表达。胃内注入生理盐水的对照组则仅有极少量的表达 ,双侧膈下迷走神经切断术可以减少c Fos蛋白在这些部位的表达。 结论　 该研究结果表明 ,迷走神经参与了胃内脏伤害性信息向孤束核及下丘脑室旁核的传递     

Induction of Fos-like protein in neurons of the medulla oblongata after electrical stimulation of the vagus nerve in anesthetized rabbit

Antibodies against the c-fos protein product Fos were used to map the first- and higher-order neurons in the rabbit medulla oblongata after electrical stimulation of the vagus nerve. Fos immunoreactivity appeared bilaterally except in the nucleus tractus solitarii. Seven areas were labeled: the nucleus tractus solitarii, the area postrema, the subnucleus lateralis caudalis magnocellularis medullar oblongata, the lateral reticular nucleus, the ambiguus nucleus, the dorsal part of the spinal trigeminal nucleus, the nucleus reticularis lateralis, the lateral border of the external cuneatus nucleus, the medial part of the inferior olivary nucleus (subnucleus β). The last two areas have never been visualized with conventional tracing techniques and may represent higher-order neurons connected to visceral vagal pathways. No labeling was observed in the nodose ganglion.     

Expression of c-fos protein in rat brain after electrical stimulation of the aortic depressor nerve

To reveal central nervous system (CNS) structures involved in the baroreceptor reflex we studied the distribution of Fos protein-like immunoreactivity in the rat brain after one hour of electrical stimulation of the aortic depressor nerve (ADN). In 13 male Wistar rats under urethane the ADN was cut on both sides and the central ends were placed on stimulating electrodes. Intermittent (11 s on, 6 s off) electrical stimulation at parameters set to elicit a drop in mean arterial pressure of 15-30 mmHg was applied to one, both or neither ADNs for 1 h. CNS sections were incubated for 48 h in anti-Fos antibody and prepared for visualization of the reaction product using the ABC immunoperoxidase technique. Label was found in several discrete brain nuclei primarily on the side ipsilateral to the side of stimulation. In the medulla labelled nuclei were found in the nucleus tractus solitarius, area postrema, rostral and caudal ventrolateral medulla, nucleus ambiguus and medullary reticular formation. In the pons labelled neurons were found in the lateral and ventrolateral parabrachial nucleus, locus coeruleus, pontine reticular field and A5 region. In the forebrain labelled nuclei were observed in the peri- and paraventricular hypothalamus, supraoptic nucleus, subfornical organ, preoptic area, central nucleus of the amygdala, median preoptic area, horizontal limb of the diagonal band, bed nucleus of the stria terminalis and islands of Calleja. In control animals moderate amounts of label were present in the supraoptic nucleus and periventricular hypothalamus bilaterally. These results define central pathways involved in mediating the baroreceptor reflex.     

Gustatory functions of the nucleus tractus solitarius in the rabbit

Multiunit analysis revealed a rostral region of NTS containing cells responsive to taste stimulation of rostral tongue. Using representative stimuli for the 4 basic types of taste, maximal incidence and magnitude of response was found to NaCl, followed by HCl, sucrose and QHC1. Further analysis of temporal patterns of response to the tastants revealed differences among stimuli in latency and time course. A principal components analysis indicated that time course, apart from magnitude of response, could contribute to neural differentiation of tastants. Information was also derived on neural intensity functions for these basic types of tastants. Additional observations with sodium saccharin revealed minimal neuronal responsivity despite reported evidence of behavioral preference for this tastant by rabbits.     

Efferent and collateral organization of paratrigeminal nucleus projections: An anterograde and retrograde fluorescent tracer study in the rat

The paratrigeminal nucleus (PTN) receives primary visceral afferent projections through cranial nerves IX and X and somatic afferent projections through cranial nerve V and dorsal roots as far caudally as C7. Pressure injections of the anterograde tracer tetramethylrhodamine dextran into the PTN in the rat resulted in bilateral labeling in the nucleus of the tractus solitarius, dorsal motor nucleus of the vagus nerve, and parabrachial nucleus. Anterograde labeling in the parabrachial nucleus was strongest in the external medial, external lateral, and ventral lateral subnuclei. Anterograde labeling was also found in the contralateral paratrigeminal nucleus, lamina I of the spinal trigeminal nucleus subnucleus caudalis, and ventroposteromedial nucleus of the thalamus. The collateral organization of PTN neurons was demonstrated by injecting different fluorescent retrograde tracers into the terminal fields of PTN projections as determined by the anterograde tracing experiments. Double-labeled neurons were found in the paratrigeminal nucleus following all combinations of injection sites. The most prominent PTN efferent projections and the most highly collateralized were to the nucleus of the tractus solitarius and parabrachial nucleus. The efferent and collateral connections of the paratrigeminal nucleus may provide a neuroanatomical substrate for integrating convergent visceral and somatic afferent information used to modulate autonomic function and behavior related to thermoregulation, nociception, and gustation. J. Comp. Neurol. 402:93–110, 1998. © 1998 Wiley-Liss, Inc.     

Autonomic and cardiovascular reflex responses to central estrogen injection in ovariectomized female rats

The role of estrogen in central autonomic nuclei was examined in ovariectomized female Sprague-Dawley rats supplemented daily for 7 days with either estrogen (5 microg/kg; sc) or saline (0.9%; sc). Animals were subsequently anaesthetized with sodium thiobutabarbital (Inactin; 100 mg/kg; ip) and instrumented to record blood pressure and heart rate. Efferent vagal parasympathetic (VPNA) and renal sympathetic (RSNA) nerve activities were recorded and used to assess baseline and reflexive changes in autonomic tone. The cardiac baroreflex was evoked using a single bolus injection of phenylephrine (0.1 mg/kg) both before and following either intrathecal injection of estrogen (0.5 microM; 1 microl) or bilateral injection of estrogen (0.5 microM; 100 nl/side) into several central autonomic nuclei. In estrogen-replaced rats, both the baseline and PE-evoked values for mean arterial pressure and RSNA were significantly decreased following injection of estrogen into the nucleus tractus solitarius (NTS), rostral ventrolateral medulla (RVLM), parabrachial nucleus (PBN), central nucleus of the amygdala (CNA) and the intrathecal space. Baseline heart rate and VPNA were significantly decreased following injection of estrogen into NTS, nucleus ambiguous (Amb), PBN and the intrathecal space. PE-evoked changes in heart rate and VPNA were significantly enhanced following injection of estrogen into these same nuclei. Injection of estrogen into the insular cortex (IC) produced significant decreases in baseline and PE-evoked RSNA only. The cardiac baroreflex was significantly enhanced following injection of estrogen into all nuclei and the intrathecal space. In saline-replaced females, injection of estrogen into NTS, RVLM, Amb and the intrathecal space had similar effects on both baseline and PE-evoked parameters although of a reduced magnitude compared to estrogen-replaced rats. However, no significant changes in autonomic tone and baroreflex function were observed following the injection of estrogen into the PBN, CNA or IC of saline-replaced rats. These results demonstrate a role for estrogen in central autonomic nuclei in female rats and suggest a possible alteration of estrogen receptor distribution or efficacy within the central nervous system of estrogen-deficient female rats.     

Axonal transport of neurotrophins by visceral afferent and efferent neurons of the vagus nerve of the rat

The receptor-mediated axonal transport of [¹²⁵I]-labeled neurotrophins by afferent and efferent neurons of the vagus nerve was determined to predict the responsiveness of these neurons to neurotrophins in vivo. [¹²⁵I]-labeled neurotrophins were administered to the proximal stump of the transected cervical vagus nerve of adult rats. Vagal afferent neurons retrogradely transported [¹²⁵I]neurotrophin-3 (NT-3), [¹²⁵I]nerve growth factor (NGF), and [¹²⁵I]neurotrophin-4 (NT-4) to perikarya in the ipsilateral nodose ganglion, and transganglionically transported [¹²⁵I]NT-3, [¹²⁵I]NGF, and [¹²⁵I]NT-4 to the central terminal field, the nucleus tractus solitarius (NTS). Vagal afferent neurons showed minimal accumulation of [¹²⁵I]brain-derived neurotrophic factor (BDNF). In contrast, efferent (parasympathetic and motor) neurons located in the dorsal motor nucleus of the vagus and nucleus ambiguus retrogradely transported [¹²⁵I]BDNF, [¹²⁵I]NT-3, and [¹²⁵I]NT-4, but not [¹²⁵I]NGF. The receptor specificity of neurotrophin transport was examined by applying [¹²⁵I]-labeled neurotrophins with an excess of unlabeled neurotrophins. The retrograde transport of [¹²⁵I]NT-3 to the nodose ganglion was reduced by NT-3 and by NGF, and the transport of [¹²⁵I]NGF was reduced only by NGF, whereas the transport of [¹²⁵I]NT-4 was significantly reduced by each of the neurotrophins. The competition profiles for the transport of NT-3 and NGF are consistent with the presence of TrkA and TrkC and the absence of TrkB in the nodose ganglion, whereas the profile for NT-4 suggests a p75 receptor-mediated transport mechanism. The transport profiles of neurotrophins by efferent vagal neurons in the dorsal motor nucleus of the vagus and nucleus ambiguus are consistent with the presence of TrkB and TrkC, but not TrkA, in these nuclei. These observations describe the unique receptor-mediated axonal transport of neurotrophins in adult vagal afferent and efferent neurons and thus serve as a template to discern the role of specific neurotrophins in the functions of these visceral sensory and motor neurons in vivo. J. Comp. Neurol. 393:102–117, 1998. Published 1998 Wiley-Liss, Inc.

1 This article is a US government work and, as such, is in the public domain in the United States of America.

     

Nicotinic acetylcholine receptor mediated modulation of evoked excitatory amino acid release in the nucleus tractus solitarius of the rat: evidence from in vivo microdialysis

In vivo microdialysis was used to measure release of endogenous -glutamate and -aspartate in the nucleus tractus solitarius of the anaesthetised rat evoked by baroreceptor loading. Aortic constriction, the method of loading, elicited a reproducible increase in extracellular levels of -glutamate to 322±139% of basal levels, which could be attenuated by concomitant local administration of the nicotinic acetylcholine receptor antagonist mecamylamine (100 μM).     

A respiratory-vocal pathway in the brainstem of the pigeon

To provide an anatomical basis for the interaction of respiration and vocalization, neuroanatomical tracing experiments were undertaken to define the link between the nucleus receiving input from the lung and the tracheosyringeal motor nucleus (nXIIts). The ventral parabrachial nucleus (PBv) receives a projection from the lateral parasolitary nucleus and from the midbrain and projects to nXIIts. The PBv is therefore well placed to gate the activity of nXIIts neurons during respiration.     

Specific subnuclei of the nucleus tractus solitarius play a role in determining the duration of inspiration in the rat

Our previous data obtained in the cat suggest that the neurons of the ventrolateral subnucleus of the tractus solitarius (vlNTS) act as an inspiratory off-switch and terminate the inspiratory phase of the respiratory cycle (Berger et al., Eur. J. Pharmacol. 277 (1995) 195–208; Gillis et al., Neurosci. Abstr. 23 (1997) 725). The purpose of the present study was to determine whether inhibition of the region of the vlNTS of the rat using drugs that hyperpolarize, disfacilitate or block both axonal conduction and action potential generation would alter the inspiratory phase of the respiratory cycle. Experiments were conducted in anesthetized, vagotomized and spontaneously breathing rats while monitoring diaphragmatic electromyogram activity. Vagus nerves were sectioned in order to rule out prolongation of inspiration evoked by microinjection of agents into the vlNTS which block excitatory drive from lung afferent inputs. Bilateral microinjection of the inhibitory amino acid γ-aminobutyric acid (GABA) 25 nmol/45 nl produced an immediate prolongation of inspiratory duration (484±18 to 1291±84 ms) and an apneustic pattern of breathing. Other effects observed were a significant shortening of expiratory duration (778±36 to 432±38 ms), rise in blood pressure (83±4 to 108±6 mmHg) and a small but significant increase in heart rate (439±17 to 452±18 beats/min). Bilateral microinjection of the ionotropic glutamate receptor antagonist kynurenic acid (1 nmol) and the Na⁺ channel blocker tetrodotoxin (10 pmol) into the region of the vlNTS consistently produced a similar prolongation of inspiratory duration and an apneustic pattern of breathing. These results support the hypothesis that neurons in the region of the vlNTS promote the transition from inspiration to expiration and function as part of the ‘Inspiratory Off Switch’.     

Effects of electrical stimulation of the glossopharyngeal nerve on cells in the nucleus of the solitary tract of the rat

Electrophysiological responses to electrical stimulation of the lingual branch of the glossopharyngeal (GP) nerve (which innervates taste buds on the caudal 1/3 of the tongue) were recorded from single cells in the rostral nucleus of the solitary tract (NTS) of anesthetized rats. Electrical stimulation was delivered as single pulses (n=55), paired-pulses (n=15) and tetanic trains (n=11). NTS cells with GP-evoked responses were also tested for responsivity to taste stimuli (0.1 M NaCl, 0.5 M sucrose, 0.01 M HCl and 0.01 M quinine HCl). Fifty-five neurons were studied: 49 cells showed GP-evoked (mean latency+/-SEM=18.0+/-1.32 ms); seven of these were taste-responsive. Spontaneous rate of these cells was low (mean+/-SEM=1.4+/-0.3 spikes per second; median=0.21 spikes per second) and many cells showed no spontaneous activity. Paired-pulse stimulation of the GP nerve in 13 rats produced both paired-pulse suppression (n=11) and paired-pulse enhancement (n=4); tetanic stimulation (25 Hz, 1.0 s) produced sustained (>20 s) increases or decreases in firing rate in 7 of 11 cells tested. Histological data suggested that GP-evoked responses recorded in the most rostral NTS were likely the result of polysynaptic connections. Cells with GP-evoked responses formed a heterogeneous group in terms of their response properties and differed from cells with evoked responses to chorda tympani (CT; which innervates taste buds on the rostral 1/3 of the tongue) nerve stimulation. These differences may reflect the respective functional specializations of the GP and CT nerves.     

Fos induction in selective hypothalamic neuroendocrine and medullary nuclei by intravenous injection of urocortin and corticotropin-releasing factor in rats

CRF and urocortin, administrated systemically, exert peripheral biological actions which may be mediated by brain pathways. We identified brain neuronal activation induced by intravenous (i.v.) injection of CRF and urocortin in conscious rats by monitoring Fos expression 60 min later. Both peptides (850 pmol/kg, i.v.) increased the number of Fos immunoreactive cells in the paraventricular nucleus of the hypothalamus, supraoptic nucleus, central amygdala, nucleus tractus solitarius and area postrema compared with vehicle injection. Urocortin induced a 4-fold increase in the number of Fos-positive cells in the supraoptic nucleus and a 3.4-fold increase in the lateral magnocellular part of the paraventricular nucleus compared with CRF. Urocortin also elicited Fos expression in the accessory hypothalamic neurosecretory nuclei, ependyma lining the ventricles and choroid plexus which was not observed after CRF. The intensity and pattern of the Fos response were dose-related (85, 255 and 850 pmol/kg, i.v.) and urocortin was more potent than CRF. Neither CRF nor urocortin induced Fos expression in the lateral septal nucleus, Edinger-Westphal nucleus, dorsal raphe nucleus, locus coeruleus, or hypoglossal nucleus. These results show that urocortin, and less potently CRF, injected into the circulation at picomolar doses activate selective brain nuclei involved in the modulation of autonomic/endocrine function; in addition, urocortin induces a distinct activation of hypothalamic neuroendocrine neurons.