A survey of oral cavity afferents to the rat nucleus tractus solitarii.

Visualization of myelinated fiber arrangements, cytoarchitecture, and projection fields of afferent fibers in tandem revealed input target selectivity in identified subdivisions of the nucleus tractus solitarii (NTS). The central fibers of the chorda tympani (CT), greater superficial petrosal nerve (GSP), and glossopharyngeal nerve (IX), three nerves that innervate taste buds in the oral cavity, prominently occupy the gustatory-sensitive rostrocentral subdivision. In addition, CT and IX innervate and overlap in the rostrolateral subdivision, which is primarily targeted by the lingual branch of the trigeminal nerve (LV). In the rostrocentral subdivision, compared with the CT terminal field, GSP appeared more rostral and medial, and IX was more dorsal and caudal. Whereas IX and LV filled the rostrolateral subdivision diffusely, CT projected only to the dorsal and medial portions. The intermediate lateral subdivision received input from IX and LV but not CT or GSP. In the caudal NTS, the ventrolateral subdivision received notable innervation from CT, GSP, and LV, but not IX. No caudal subnuclei medial to the solitary tract contained labeled afferent fibers. The data indicate selectivity of fiber populations within each nerve for functionally distinct subdivisions of the NTS, highlighting the possibility of equally distinct functions for CT in the rostrolateral NTS, and CT and GSP in the caudal NTS. Further, this provides a useful anatomical template to study the role of oral cavity afferents in the taste-responsive subdivision of the NTS as well as in subdivisions that regulate ingestion and other oromotor behaviors.

Chronic hypoxia upregulates connexin43 expression in rat carotid body and petrosal ganglion.

Recent studies have demonstrated that oxygen-sensitive type I cells in the carotid body express the gap junction-forming protein connexin43 (Cx43). In the present study, we examined the hypothesis that chronic exposure to hypoxia increases Cx43 expression in type I cells as well as in chemoafferent neurons in the petrosal ganglion. Immunocytochemical studies in tissues from normal rats revealed diffuse and granular Cx43-like immunoreactivity in the cytoplasm of type I cells and dense punctate spots of immunoreactive product at the margins of type I cells and near the borders of chemosensory cell lobules. Cx43-like immunoreactivity was not detectable in petrosal ganglion neurons from normal animals. After a 2-wk exposure to hypobaric (380 Torr) hypoxia, Cx43 immunostaining was substantially enhanced in and around type I cells. Moreover, chronic hypoxia elicited the expression of Cx43-like immunoreactivity in the cytoplasm of afferent neurons throughout the petrosal ganglion. Quantitative RT-PCR studies indicate that chronic hypoxia evokes a substantial increase in Cx43 mRNA levels in the carotid body, along with a marked elevation of Cx43 expression in the petrosal ganglion. Increased Cx43 expression and gap junction formation in type I cells and sensory neurons may contribute to carotid body adaptation during sustained stimulation in extreme physiological conditions.

5-HT5a receptors in the carotid body chemoreception pathway of rat.

By using a specific antibody, 5-HT5a receptor-like immunoreactivity was revealed in the chemoreceptive, oxygen sensitive, carotid body (CB) type I cells, and neurons of the petrosal ganglion (PG) and the superior cervical ganglion (SCG) in rat. mRNA encoding for the 5-HTa receptor was also detected in these tissues by RT-PCR, and confirmed with DNA sequencing. The present study provides direct evidence that 5-HT5a receptors are expressed in the CB, PG and SCG, which all likely play fundamental roles in arterial chemoreception.

The coexistence of TrkA with putative transmitter agents and calcium-binding proteins in the vagal and glossopharyngeal sensory neurons of the adult rat.

The presence of the neurotrophin receptor, TrkA, in neurochemically identified vagal and glossopharyngeal sensory neurons of the adult rat was examined. TrkA was colocalized with calcitonin gene-related peptide (CGRP), parvalbumin, or calbindin D-28k in neurons of the nodose, petrosal and/or jugular ganglia. In contrast, no TrkA-immunoreactive (ir) neurons in these ganglia colocalized tyrosine hydroxylase-ir. About one-half of the TrkA-ir neurons in the jugular and petrosal ganglia contained CGRP-ir, whereas only a few of the numerous TrkA-ir neurons in the nodose ganglion contained CGRP-ir. Although 43% of the TrkA-ir neurons in the nodose ganglion contained calbindin D-28k-ir, few or no TrkA-ir neurons in the petrosal or jugular ganglia were also labeled for either calcium-binding protein. These data show distinct colocalizations of TrkA with specific neurochemicals in vagal and glossopharyngeal sensory neurons, and suggest that nerve growth factor (NGF), the neurotrophin ligand for TrkA, plays a role in functions of specific neurochemically defined subpopulations of mature vagal and glossopharyngeal sensory neurons.

Normal ontogenic observations on the expression of Eph receptor tyrosine kinase, Cek8, in chick embryos.

The spatio-temporal pattern of expression for the Eph receptor tyrosine kinase, Cek8, was observed in normal chick embryos from H-H stage 6 to 23 by immunohistochemical techniques. Expression of Cek8 was already present in embryos at H-H stage 6, where it was located in the neural plate of the brain region, paraxial mesoderm, and the primitive streak. Regions expressing Cek8 subsequently increased during development to include the neural folds of the brain, rhombomeres 3 and 5, the caudal part of the neural plate, neural crest cells related to the formation of glossopharyngeal nerve ganglia, invaginated cells throughout the primitive groove and the epithelium of the rudiment of the gall bladder. Cek8 was also expressed in the mesenchymal cells of the pharyngeal arches, allantoic stalk and limb buds as well as in the areas surrounding the eye vesicles and nasal pits. Furthermore, cells in the tail bud progressing to the secondary neurulation expressed Cek8. Thus, the spatiotemporal patterns of Cek8 expression appears to have intimate relationships with tissue rebuilding, the maturation of differentiated cells, and the spatial organization of tissues. Consequently, it appears that Cek8 plays an integral role in the developmental events leading to the formation of a wide--though not inclusive--variety of tissues and organ systems.

Coexistence of calcium-binding proteins in vagal and glossopharyngeal sensory neurons of the rat.

The presence and coexistence of the calcium-binding proteins (CaBPs), calbindin D-28k, parvalbumin and S100 protein, were immunohistochemically examined in the glossopharyngeal and vagal sensory ganglia, the carotid body and taste buds. The CaBPs were found in each ganglion with the nodose ganglion containing the largest number of CaBP-immunoreactive (ir) cells (calbindin D-28k > or = S100 >> parvalbumin). The coexistence of CaBPs was found in neurons of the nodose, petrosal, and jugular ganglia. Calbindin D-28k-ir neurons in the nodose and petrosal ganglia frequently colocalized S100-ir whereas calbindin D-28k-ir neurons in the jugular ganglion less frequently contained S100-ir. Only small percentages of calbindin D-28k-ir neurons in each ganglion colocalized parvalbumin. Similarly, S100-ir neurons in the nodose and petrosal ganglia frequently colocalized calbindin D-28k-ir whereas S100-ir neurons in the jugular ganglion less frequently contained calbindin D-28k-ir. Moderate to small percentages of S100-ir neurons in each ganglion colocalized parvalbumin. Parvalbumin-ir neurons nearly always colocalized S100-ir in the nodose, petrosal and jugular ganglia. Moderate to small percentages of parvalbumin-ir neurons in each ganglion colocalized calbindin D-28k. Whereas calbindin D-28k- and S100-ir were colocalized in nerve fibers and cells within taste buds of circumvallate papilla of the tongue, the coexistence of these CaBPs could not be determined in the carotid body. These findings suggest a co-operative role for CaBPs in the functions of subpopulations of nodose and petrosal ganglia neurons.

Distribution of neuropeptide-immunoreactive nerve fibres in the medulla oblongata of the chicken.

The anatomical distribution of nerve fibres containing galanin (GAL), leucine enkephalin (lENK), methionine enkephalin (mENK) and substance P (SP) was investigated in the medulla oblongata of the chicken by means of immunohistochemistry. In the medulla oblongata, dense networks of mENK-, lENK-, SP- and GAL-immunoreactive nerve fibres were mainly restricted within and around the dorsal vagal complex, i.e., the nucleus of the solitary tract and dorsal motor nucleus of the vagus nerve. The relative density of immunoreactive nerve fibres was usually decreased in the order of mENK-, SP-, lENK- and GAL-immunoreactive ones. Immunoreactive nerve fibres for each antiserum had a unique distribution among the subnuclear groups of the dorsal vagal complex. Sparser networks of immunolabelled nerve fibres were found in the nucleus of the hypoglossal nerve, nucleus ambiguus, nucleus of the descending trigeminal tract, raphe nucleus and caudal olivary complex. In the colchicine-pretreated chicken, some nerve cell bodies expressing the immunoreaction for ENK and GAL antisera were detected in the nucleus of the solitary tract and nucleus of the descending trigeminal tract. There were some differences in the distribution pattern of immunoreactive nerve elements among the four kinds of antisera. The selective distribution of neuropeptide-containing nerve elements within various nuclei of the medulla oblongata suggests that these neuropeptides may be differentially involved in central neural regulation.

Presence and localization of neurotrophin receptor tyrosine kinase (TrkA, TrkB, TrkC) mRNAs in visceral afferent neurons of the nodose and petrosal ganglia.

The presence of mRNAs to the high affinity tyrosine kinase (Trk) receptors for neurotrophins was studied in visceral afferent neurons of the nodose and petrosal ganglia of adult and neonatal rats using in situ hybridization histochemistry. Neurons containing TrkA mRNA were found in the adult nodose and petrosal ganglia. About 10% of nodose ganglion neurons and 38% of petrosal ganglion neurons contained TrkA mRNA. The nodose and petrosal ganglia from 1 day old neonates also expressed TrkA mRNA. No TrkB mRNA-containing neurons were detected in the adult nodose and petrosal ganglia, whereas TrkB mRNA was detected in 1 day old neonatal nodose and petrosal ganglia. TrkC mRNA was found in about 9% of nodose ganglion neurons and 11% of petrosal ganglion neurons of adult rats. Likewise, low but detectable levels of TrkC mRNA were seen in 1 day old neonatal nodose and petrosal ganglia. These data demonstrate the presence of TrkA and TrkC in the adult nodose and petrosal ganglia and provide a substrate for the ongoing neurotrophin-induced regulation of these placodally derived visceral afferent neurons. The altered expression of Trk receptor mRNAs in the nodose and petrosal ganglia between the adult and neonatal rats may reflect developmentally regulated changes in neurotrophin responsiveness.

Ultrastructure of calcitonin gene-related peptide-immunoreactive, unmyelinated afferents to the cat carotid body: a case of volume transmission.

To relate the ultrastructure of unmyelinated afferents to the cat carotid body with the known electrophysiological properties of cat chemosensory C-fibers, we took advantage of the fact that the calcitonin gene-related peptide is exclusively present in a population of sparsely branched afferents to the carotid body. They have a morphology identical to the afferents originating from carotid sinus nerve unmyelinated axons. Immunoreactive axons were stained using pre-embedding protocols and horseradish peroxidase-labeled secondary antibody. Labeling was present only in unmyelinated axons and boutons distributed in the interstitial and parenchymal tissue. The varicosities had an average diameter of 0.7 micron, and contained both small, clear vesicles and larger dense-core vesicles. No labeled axons were ever seen to contact glomus cells, but could be observed as close as 0.2 micron to a glomus cell, always with an interposed glial process. With a very sensitive protocol, that used tungstate-stabilized tetramethylbenzidine as the chromogen, amorphous deposits of reaction product were often detected in the extracellular space around a labeled bouton. We interpret these findings as indicating that the reciprocal chemical transmission between the oxygen-sensitive glomus cells and the unmyelinated afferents takes place through non-synaptic transmission, via the rather large extracellular space of the carotid body. In addition, the larger distances between glomus cells and unmyelinated afferents could explain the lowered sensitivity and sluggishness of chemosensory C-fibers, compared to the A-fibers.