Dissection and surgical approaches to the mouse jugular-nodose ganglia.

The jugular-nodose ganglia contain the sensory peripheral neurons of the vagus nerve, linking visceral organs to the medulla oblongata. Accessing these ganglia in smaller animals without damaging the vascular and neural structures may be challenging, as ganglionic fibers imbed deeply into the carotid sheath, and vagal parasympathetic fibers cross through the interior of the ganglia. We describe a practical protocol for locating and accessing the mouse jugular-nodose ganglia in vivo, including instructions for intraganglionic injections and postperfusion dissection. For complete details on the use and execution of this protocol, please refer to Han et al. (2018).

Characterization of a cell bridge variant connecting the nodose and superior cervical ganglia in the mouse: Prevalence, anatomical features, and practical implications.

While autonomic ganglia have been extensively studied in rats instead of mice, there is renewed interest in the anatomy of the mouse autonomic nervous system. This study examined the prevalence and anatomical features of a cell bridge linking two autonomic ganglia of the neck, namely, the nodose ganglion (NG) and the superior cervical ganglion (SCG) in a cohort of C57BL/6J mice. We identified a cell bridge between the NG and the cranial pole of the SCG. This cell bridge was tubular shaped with an average length and width of 700 and 240 µm, respectively. The cell bridge was frequently unilateral and significantly more prevalent in the ganglionic masses from males (38%) than females (21%). On each of its extremities, it contained a mixed of vagal afferents and postganglionic sympathetic neurons. The two populations of neurons abruptly replaced each other in the middle of the cell bridge. We examined the mRNA expression for selected autonomic markers in samples of the NG with or without cell bridge. Our results indicated that the cell bridge was enriched in both markers of postganglionic sympathetic and vagal afferents neurons. Lastly, using FluoroGold microinjection into the NG, we found that the existence of a cell bridge may occasionally lead to the inadvertent contamination of the SCG. In summary, this study describes the anatomy of a cell bridge variant consisting of the fusion of the mouse NG and SCG. The practical implications of our observations are discussed with respect to studies of the mouse vagal afferents, an area of research of increasing popularity.

Causes and consequences of degeneration of the dorsal motor nucleus of the vagus nerve in Parkinson's disease.

SIGNIFICANCE: Parkinson's disease (PD) is no longer considered merely a movement disorder caused by degeneration of dopamine neurons in the midbrain. It is now recognized as a widespread neuropathological syndrome accompanied by a variety of motor and nonmotor clinical symptoms. As such, any hypothesis concerning PD pathogenesis and pathophysiology must account for the entire spectrum of disease and not solely focus on the dopamine system. RECENT ADVANCES: Based on its anatomy and the intrinsic properties of its neurons, the dorsal motor nucleus of the vagus nerve (DMV) is uniquely vulnerable to damage from PD. Fibers in the vagus nerve course throughout the gastrointestinal (GI) tract to and from the brainstem forming a close link between the peripheral and central nervous systems and a point of proximal contact between the environment and areas where PD pathology is believed to start. In addition, DMV neurons are under high levels of oxidative stress due to their high level of α-synuclein expression, fragile axons, and specific neuronal physiology. Moreover, several consequences of DMV damage, namely, GI dysfunction and unrestrained inflammation, may propagate a vicious cycle of injury affecting vulnerable brain regions. CRITICAL ISSUES: Current evidence to suggest the vagal system plays a pivotal role in PD pathogenesis is circumstantial, but given the current state of the field, the time is ripe to obtain direct experimental evidence to better delineate it. FUTURE DIRECTIONS: Better understanding of the DMV and vagus nerve may provide insight into PD pathogenesis and a neural highway with direct brain access that could be harnessed for novel therapeutic interventions.

Anatomical anomalies of the laryngeal branches of the vagus nerve in pigs (Sus scrofa).

To delineate the anomaly and frequency of their occurrence in a pig model, we reported the topography of the vagus laryngeal branches and compared the differences with humans. Thirty sides of cervical vagus nerve in 15 fresh cadavers (Sus scrofa) were microdissected. We measured the branch diameters and lengths of the laryngeal branches using a Vernier caliper with a resolution of 0.01 mm. Two patterns of the vagus laryngeal branches were shown: 56.7% with the cranial laryngeal nerve (CLN) and 43.3% without the CLN. The diameters and the length of the CLN were not affected by the side of the neck (P > 0.05), but the diameters of the recurrent laryngeal nerve (RLN) and the nodose ganglion were significantly different between left and right sides (P < 0.05). The left RLN was thinner than the right side in diameter (P < 0.05). Four of the 30 sides had anastomoses between the vagus and the cervical sympathetic chain. There were some differences between the pig anatomy and human anatomy, but the patterns were largely similar. The similarities support the utility of this model, which is closer in size to humans than the standard rodent models.

Selective expression of a sodium pump isozyme by cough receptors and evidence for its essential role in regulating cough.

We have identified a distinct subtype of airway vagal afferent nerve that plays an essential role in regulating the cough reflex. These afferents are exquisitely sensitive to punctate mechanical stimuli, acid, and decreases in extracellular chloride concentrations, but are insensitive to capsaicin, bradykinin, histamine, adenosine, serotonin, or changes in airway intraluminal pressures. In this study we used intravital imaging, retrograde neuronal tracing, and electrophysiological analyses to characterize the structural basis for their peculiar mechanical sensitivity and to further characterize the regulation of their excitability. In completing these experiments, we uncovered evidence for an essential role of an isozyme of Na(+)-K(+) ATPase in regulating cough. These vagal sensory neurons arise bilaterally from the nodose ganglia and are selectively and brilliantly stained intravitally with the styryl dye FM2-10. Cough receptor terminations are confined and adherent to the extracellular matrix separating the airway epithelium and smooth muscle layers, a site of extensive remodeling in asthma and chronic obstructive pulmonary disease. The cough receptor terminals uniquely express the alpha(3) subunit of Na(+)-K(+) ATPase. Intravital staining of cough receptors by FM2-10, cough receptor excitability in vitro, and coughing in vivo are potently and selectively inhibited by the sodium pump inhibitor ouabain. These data provide the first detailed morphological description of the peripheral terminals of the sensory nerves regulating cough and identify a selective molecular target for their modulation.

Morphology and topography of nucleus ambiguus projections to cardiac ganglia in rats and mice.

Vagal efferent axons from the nucleus ambiguus (NA) innervate ganglionated plexuses in the dorsal surface of cardiac atria, which in turn, may have different functional roles in cardiac regulation. However, the morphology and topography of vagal efferent projections to these ganglionated plexuses in rats and mice have not been well delineated. In the present study, we injected the tracer 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine methanesulfonate (DiI) into the left NA to label vagal efferent axons and terminals in cardiac ganglia and administered Fluoro-Gold (FG) i.p. to stain cardiac ganglia. Then, we used confocal microscopy and a Neurolucida 3-D Digitization System to qualitatively and quantitatively examine the distribution and structure of cardiac ganglia, and NA efferent projections to cardiac ganglia in the whole-mounts of Sprague-Dawley (SD) rats and FVB mice. Our observations were: 1) Cardiac ganglia of different shapes and sizes were distributed in the sinoatrial (SA) node, atrioventricular (AV) node, and lower pulmonary vein (LPV) regions on the dorsal surface of the atria. In each region, several ganglia formed a ganglionated plexus. The plexuses at different locations were interconnected by nerves. 2) Vagal efferent fibers ramified within cardiac ganglia, formed a complex network of axons, and innervated cardiac ganglia with very dense basket endings around individual cardiac principal neurons (PNs). 3) The percent of the PNs in cardiac ganglia which were innervated by DiI-labeled axons was 54.3+/-3.2% in mice vs. 53.2+/-3.2% in rats (P>0.10). 4) The density of axonal putative-synaptic varicosities on the surface of PNs was 0.15+/-0.02/microm(2) in mice vs. 0.16+/-0.02/microm(2) in rats (P>0.10). Thus, the distributions of cardiac ganglia and vagal efferent projections to cardiac ganglia in mice and rats were quite similar both qualitatively and quantitatively. Our study provides the structural foundation for future investigation of functional differentiation of ganglionated plexuses and the brain-heart circuitry in rodent models of human disease.

Nitroxidergic neurons in rat nucleus tractus solitarii express vesicular glutamate transporter 3.

Earlier we reported that glutamate transporter (VGLUT) 2 and neuronal nitric oxide synthase (nNOS) are colocalized in some fibers and are present in apposing fibers in the nucleus tractus solitarii (NTS). Those findings provided anatomical support for a hypothesized physiological link between glutamate and nitric oxide (NO.) in the NTS. Recently a third class of VGLUT, VGLUT3, was identified, but its distribution in NTS and its anatomical relationship with nNOS have not been shown. In this study we tested the hypothesis that neurons and fibers containing VGLUT3 lie in close proximity to those containing nNOS and that both proteins colocalize in some neurons and fibers in the NTS. We perfused rats and obtained brain stem sections and nodose ganglion sections for immunofluorescent staining analyzed by confocal microscopy. The NTS contained moderate VGLUT3-immunoreactivity (IR), with the intermediate, medial and interstitial subnuclei containing higher VGLUT3-IR than other subnuclei. Although all three forms of VGLUT were present in the NTS, VGLUT3-IR was not colocalized with either VGLUT1-IR or VGLUT2-IR in either processes or cells in the brain stem. Cells and processes containing both VGLUT3-IR and nNOS-IR were noted in all NTS subnuclei and in the nodose ganglion. Triple immunofluorescent staining revealed that cells double-labeled for nNOS-IR and VGLUT3-IR were all additionally labeled for neuronal nuclear antigen (NeuN), a neuronal marker. These findings support our hypothesis that neurons and fibers containing VGLUT3 lie in close proximity to those containing nNOS and that both proteins colocalize in some neurons and fibers in the NTS.

[Morphometric characteristics of rat viscero- and somatosensory neurocytes of the caudal ganglia in the vagus nerve]. / Morfometricheskaia kharakteristika vistsero- i somatosensornykh neirotsitov kaudal'nogo uzla bluzhdaiushchego nerva krysy.

In experiments performed in 63 adult albino male rats, the participation of afferent neurons of caudal vagal ganglion in the innervation of different organs was investigated. Horseradish peroxidase and conjugate of horseradish peroxidase with wheat germ agglutinin were used as the tracers. Metric parameters (diameter of an equivalent circle) and shape parameters (circular factor of the form) of marked neurocytes were studied using computer videoanalysis method. The dependence of neuronal cell body dimensions on the distance to the innervated target organ was established. The exclusion was represented by the neurocytes, innervating the root of the tongue, which are larger than those projecting to the pancreas. The reason for these discrepancies seems to be associated with the differences of viscero- and somatosensory relations.

Postnatal development of hypothalamic inputs to the dorsal vagal complex in rats.

The hypothalamus is critically involved in energy homeostasis and is an appropriate focus for research investigating the central neural underpinnings of obesity, anorexia and normal food intake. However, little is known regarding pathways and mechanisms that convey relevant hypothalamic signals to the brainstem circuits that ultimately control ingestive behavior. This brief review highlights work investigating the postnatal development of hypothalamic inputs to the hindbrain dorsal vagal complex (DVC). Research findings indicate that these inputs are both structurally and functionally immature in newborn rats. The progressive postnatal maturation of descending projections to the DVC occurs in concert with newly emerging physiological and behavioral responses to osmotic dehydration, which inhibits gastric emptying and food intake in adult animals but not in neonates. The postnatal emergence of other intake controls might also reflect progressive engagement of DVC neural circuits, whose intrinsic components and output pathways are envisioned as being critical for initiating and terminating ingestive behavior.

[Projections of the gastrointestinal tract organs on afferent ganglions of the vagus nerve in rats]. / Proektsiia organov zheludochno-kishechnogo trakta na afferentnye uzly bluzhdaiushchego nerva krysy.

Experiments were carried in 42 mature white male rats. We investigated into the projections of different organs of gastrointestinal tract on afferent neurones of ganglia of the vagus nerve in white rats. Horseradish peroxidase-conjugated lectins were used as a tracer. We investigated into metric parameters (diameter of an equivalent circle) and shape parameters (circular factor of the shape) of marked neurocytes using a method of computer video-analysis. To evaluate reliability of the received data, methods of non-parametric statistics were used. It was determined that the largest neurocytes in afferent ganglions are involved in innervation of the root of the long and ileocecal angle, the smallest ones--in innervation of a cervical department of esophagus and liver. The viscero- and somatosensory neurocytes in caudal ganglion of vagus nerve involved in innervation of different organs in white rat, are characterized by selectivity of the shape and by metric parameters.

[Populations of afferent neurons in the sensory ganglia detected using lectin from Laburnum anagyroides]. / Populiatsii afferentnykh neironov v chuvstvitel'nykh uzlakh, vyiavlennye s ispol'zovaniem lektina bobovnika anagirolistnogo.

The purpose of the present research was the study of afferent neuron subpopulations in vagal caudal ganglia and trigeminal ganglion of adult albino rats using a conjugate of fucose-specific Laburnum anagyroides lectin (LAL) with peroxidase. Histochemical preparations obtained were examined using computer videoanalyzer with the determination of dimensions of afferent neurons and integral optical density (IOD) of their cytoplasm. In the ganglia studied LAL was found to bind to the majority (more than 98%) of neurons. Reaction product was demonstrated in the perikaryon either as discrete granules (Nissl body-type), or as a uniform precipitate. Nucleoplasm in most of neurons remained lightly stained. Determination of IOD of neuronal cytoplasm in various ganglia demonstrated significant differences in degree of LAL accumulation. Analysis of interrelation between neuronal size and IOD permitted to establish non-linear correlation of metric and optical parameters and to detect subpopulations of cells in the sensory ganglia, which were stained with LAL most intensely. Functional specialization of these cells remains to be determined. Thus, a combined application of lectin-histochemical method with a computer videoanalysis of morphological slides enabled the identification neuronal populations and subpopulations in rat afferent ganglia, which are not demonstrated with the standard histological methods.

Nerve growth factor-induced phenotypic switch in guinea pig airway sensory neurons.

Immunohistochemistry was combined with retrograde tracing techniques to characterize the effect of nerve growth factor (NGF) on substance P (SP) producing vagal neurons innervating the guinea pig trachea. Fast blue dye instilled into the trachea retrogradely labeled nerve cell bodies located in the nodose and jugular ganglia. In untreated guinea pigs > 99% of the SP-containing neurons labeled with fast blue were located in the jugular ganglia. The SP-positive neurons were small in diameter (23 +/- 1 microm) and were negative for neurofilament immunoreactivity. The fast-blue-positive neurons in the nodose ganglia, by contrast, were large in diameter (40 +/- 3 microm) and were negative for SP immunoreactivity and positive for neurofilament immunoreactivity. After NGF-beta injections into the tracheal wall, approximately 10% of the large-diameter nodose neurofilament-positive neurons projecting fibers to the trachea became SP-positive (p < 0.05). We previously demonstrated that nodose nerve endings supplying the trachea are exquisitely mechanically sensitive, but capsaicin- and bradykinin-insensitive. These results suggest that NGF not only increases SP expression in airway neurons, but changes the neuronal phenotype such that large, capsaicin-insensitive nodose neurons with fast-conducting "Adelta" fibers provide a component of the tachykinergic innervation.

Morphology and development of the distal ganglion of the vagus nerve (ganglion distale n. vagi) in sheep during the prenatal period.

This study was carried out on 31 sheep fetuses, which were divided into four developmental groups. It was established that the development and morphology of the investigated part of vagus nerve is strongly correlated with the developing organs of the fetuses. The very distinct 'translocation' of the distal ganglion of the vagus nerve in the caudoventral direction was observed in the course of development. There was no influence of sex on the results of investigation.

Neurochemistry of the nodose ganglion.

Placode-derived general visceral afferent neurons of the nodose ganglion transmit visceral sensory information from specialized sensory endings of the vagus nerve and its branches to the nucleus of the solitary tract. These neurons are critical in relaying information such as elevations in blood pressure, changes in blood oxygenation, passage of contents through the esophagus and intestines, and distention of the heart, stomach, and lungs to the CNS for reflex maintenance of visceral functions. Multiple neurotransmitters, neuropeptides, calcium binding proteins, and other neuroactive substances are associated with neurons of the nodose ganglion. Many neurons colocalize 2 or more neuroactive substances creating the potential for complex interactions of neurochemical signals in the NTS. Neurons of the nodose ganglion also contain a variety of receptors which respond to transmitters, inflammatory mediators, and neurotrophic factors. The contents of these neurochemicals and receptors are not static as alterations in their expression are noted in response to epigenetic influences. Although not yet well understood, potential factors and mechanisms regulating neurochemical events in the nodose ganglion neurons are discussed.

Vagal afferent innervation of rat abdominal paraganglia as revealed by anterograde DiI-tracing and confocal microscopy.

Abdominal vagal afferent fibers were selectively labeled by injecting the fluorescent carbocyanine dye DiI into the left nodose ganglion of rats. Almost all paraganglia that were distributed along the five major abdominal vagal branches and their subbranches were found to be innervated by labeled vagal afferents. Laser scanning confocal microscopy with its single optical sectioning and three-dimensional reconstruction capabilities were used to analyze this innervation in more detail for paraganglia near the vagal hepatic branch and liver hilus. Furthermore, in double-labeling studies, it was demonstrated that a large percentage of the vagally innervated glomus cells were capable of catecholamine synthesis on the basis of their positive staining for tyrosine hydroxylase antibody. These findings support the concept of a chemoreceptive function for the abdominal paraganglia.

Visualization of dopamine D2 binding sites on human inferior vagal ganglia.

The present study used in vitro autoradiography to assess whether high affinity D2 binding sites are present on sections of human inferior vagal ganglia, where arterial baroreceptor primary afferent cell bodies are located. Incubation of tissue sections with [125I]NCQ 298 (0.5 nM) revealed dense but non-uniform binding, with a well-defined topography, consistent with the localization of binding sites over cell bodies rather than nerve axons. Coincubation with the D2 antagonist raclopride (10 microM) completely abolished binding of [125I]NCQ 298. Densitometric quantification of autoradiograms estimated 82 +/- 4% specific binding for [125I]NCQ 298 (0.5 nM). These observations indicate the presence of D2 binding sites on sensory ganglia involved in cardiovascular control pathways in the human, and may help to explain some of the documented cardiovascular effects of dopamine.

Sensory ganglia as a target of autonomic and sensory nerve fibres in the guinea-pig.

The distribution of neuropeptide- (neuropeptide Y, substance P, vasoactive intestinal peptide) and catecholamine-synthesizing enzyme-immunoreactive axons in guinea-pig trigeminal, nodose, and cervical dorsal root ganglia was studied by double-labelling immunofluorescence in controls and after extirpation of either the cervical sympathetic trunk or the stellate ganglion; tyrosine hydroxylase- and dopamine-beta-hydroxylase-immunoreactive terminals in dorsal root ganglia were ultrastructurally investigated. Six neurochemically identifiable axons innervated the trigeminal ganglion, five kinds were found in the nodose and dorsal root ganglia. Two of them (catecholaminergic with and without neuropeptide Y) were of sympathetic origin and, besides their termination at arteries, provided a direct innervation of capsule cells of the trigeminal and cervical dorsal root ganglia facing the subarachnoid space. Varicosities which were interpreted as being of sensory origin were equally numerous in all ganglia, whereas those being likely of parasympathetic origin decreased in numbers from the trigeminal to the dorsal root and nodose ganglia. It is concluded that the sensory ganglia are the target of postganglionic sympathetic, parasympathetic and primary afferent neurons, each of which are specifically organized with respect to the neurochemical phenotype and inter- and intraganglionic distribution. Among other targets, these "nervi gangliorum" appear to be intimately linked to the ganglionic capsular cells and meningeal sheaths facing the liquor spaces.

[The morphological characteristics of the parasympathetic innervation of the pyloric sphincter in the cat]. / Morfologicheskie osobennosti parasimpaticheskoi innervatsii piloricheskogo sfinktera u koshki.

The method of axonal transport of horse-radish peroxidase was used to detect the localization of neurons in the dorsal motor nucleus of the vagus nerve sending the axons to the pyloric sphincter. The investigation was carried out in cats. Under study were also morphological features of the nodular ganglion responsible for afferent innervation of the sphincter. The maximum amount of the corresponding cells are found in the dorsomedial part of the dorsal motor nucleus in the area from +1.0 to +2.0 mm (with respect to obex). The afferent neurons to which information comes from interoceptors of the sphincter zone along the vagus nerve fibers, are distributed in the left and right nodular ganglia almost evenly. The major part of these cells have the area of 300-800 mkm2.