Calcium-fluxing glutamate receptors associated with primary gustatory afferent terminals in goldfish (Carassius auratus).

Presynaptic ionotropic glutamate receptors modulate transmission at primary afferent synapses in several glutamatergic systems. To test whether primary gustatory afferent fibers express Ca(2+)-permeable AMPA/kainate receptors, we utilized kainate-stimulated uptake of Co(2+) along with immunocytochemistry for the Ca(2+)-binding proteins (CaBPs) calbindin and calretinin to investigate the primary gustatory afferents in goldfish (Carassius auratus). In goldfish, the primary gustatory nucleus (equivalent to the gustatory portion of the nucleus of the solitary tract) includes the vagal lobe, which is a large, laminated structure protruding dorsally from the medulla. Kainate-stimulated uptake of Co(2+) (a measure of Ca(2+)-fluxing glutamate receptors) shows punctate staining distributed in the distinct laminar pattern matching the layers of termination of the primary gustatory afferent fibers. In addition, CaBP immunocytochemistry, which correlates highly with expression of Ca(2+)-permeable AMPA/kainate receptors, shows a laminar pattern of distribution similar to that found with kainate-stimulated cobalt uptake. Nearly all neurons of the vagal gustatory ganglion show Co(2+) uptake and are immunopositive for CaBPs. Transection of the vagus nerve proximal to the ganglion results in loss of such punctate Co(2+) uptake and of punctate CaBP staining as soon as 4 days postlesion. These results are consonant with the presence of Ca(2+)-fluxing glutamate receptors on the presynaptic terminals of primary gustatory terminals, providing an avenue for modulation of primary gustatory input.

Trigeminal collaterals in the nasal epithelium and olfactory bulb: a potential route for direct modulation of olfactory information by trigeminal stimuli.

The nasal epithelium is richly invested with peptidergic (substance P and calcitonin gene-related peptide [CGRP]) trigeminal polymodal nociceptors, which respond to numerous odorants as well as irritants. Peptidergic trigeminal sensory fibers also enter the glomerular layer of the olfactory bulb. To test whether the trigeminal fibers in the olfactory bulb are collaterals of the epithelial trigeminal fibers, we utilized dual retrograde labeling techniques in rats to identify the trigeminal ganglion cells innervating each of these territories. Nuclear Yellow was injected into the dorsal nasal epithelium, and True Blue was injected into the olfactory bulb of the same side. Following a survival period of 3-7 days, the trigeminal ganglion contained double-labeled, small (11.8 x 8.0 microm), ellipsoid ganglion cells within the ethmoid nerve region of the ganglion. Tracer injections into the spinal trigeminal complex established that these branched trigeminal ganglion cells also extended an axon into the brainstem. These results indicate that some trigeminal ganglion cells with sensory endings in the nasal epithelium also have branches reaching directly into both the olfactory bulb and the spinal trigeminal complex. These trigeminal ganglion cells are unique among primary sensory neurons in having two branches entering the central nervous system at widely distant points. Furthermore, the collateral innervation of the epithelium and bulb may provide an avenue whereby nasal irritants could affect processing of coincident olfactory stimuli.

Distribution of cholecystokinin, calcitonin gene-related peptide, neuropeptide Y, and galanin in the primary gustatory nuclei of the goldfish.

Cholecystokinin (CCK), neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP), and galanin all are known to have central effects on food intake. Immunocytochemistry was used to examine the presence of these substances within the primary gustatory nuclei of the goldfish, including the vagal lobe, which is a large, laminated structure composed of discrete sensory, fiber, and motor layers. The vagal lobes receive primary afferent input from the gustatory portion of the vagus nerve and contain reflex circuitry involved in the ingestion or rejection of potential food items. Immunohistochemistry indicates a heavy concentration of CCK-, CGRP-, NPY-, and galanin-immunoreactive fibers in the capsular fiber layer as well as in deeper sensory layers of the vagal lobe. CGRP immunoreactivity throughout the sensory layers and capsular immunoreactivity for CCK are greatly reduced 1-2 weeks following vagus nerve transection, indicating that the majority of these fibers are primary sensory afferents. In contrast, NPY and galanin immunoreactivity in the capsular fiber layer and reactivity for CCK, NPY, and galanin in the deeper sensory and fiber layers are relatively unaffected by vagus transection. CCK-, NPY-, and galanin-immunoreactive fibers and puncta also were present in the motor layers, as were CGRP-immunoreactive motor somata. CCK-immunoreactive cell bodies are present in layer III and layer VII/VIII of the vagal lobe and in the superficial granular layer of the lateral subnucleus of the commissural nucleus of Cajal, which is caudally contiguous with the vagal lobe.

Neurotrophin-3 is expressed in a discrete subset of olfactory receptor neurons in the mouse.

In transgenic neurotrophin-3 lacZ-neo (NT-3(lacZneo)) mice, in which the coding region for NT-3 is replaced by Eschericia coli lacZ, the expression of beta-galactosidase faithfully mimics the expression of NT-3 (Vigers AJ, Baquet ZC, Jones KR [2000], J Comp Neurol 416:398-416). During embryonic and early postnatal development, beta-galactosidase is detected in the olfactory system, beginning at embryonic day 11.5 in the nasal epithelium and at embryonic day 16.5 in the olfactory bulb. Levels of beta-galactosidase rise with age, reaching a peak during the second postnatal week, when beta-galactosidase reactivity is visible in up to 50% of the glomeruli. As the animal matures, the beta-galactosidase levels decline, but staining remains present in axons and cell bodies of a specific subset of olfactory receptor neurons (ORNs) projecting to a limited subset of glomeruli. The heavily labeled ORNs do not follow the typical OR expression zones in the epithelium but appear similar to the "patch" expression pattern of mOR37 receptors. The most heavily reactive glomeruli exhibit a striking reproducible pattern in the ventral olfactory bulb (OB). Some glomeruli of the OB contain calcitonin gene-related peptide (CGRP)-immunoreactive fibers of the trigeminal nerve. However, double-label immunocytochemistry for CGRP and beta-galactosidase rendered no correlation between trigeminal innervation and the degree of innervation by NT-3-expressing ORNs. Thus, the timing and presence of beta-galactosidase in a subset of ORNs suggests that NT-3 plays a role in synaptogenesis and/or synapse function in a specific subset of ORNs within the olfactory bulb.

Solitary chemoreceptor cells in the nasal cavity serve as sentinels of respiration.

Inhalation of irritating substances leads to activation of the trigeminal nerve, triggering protective reflexes that include apnea or sneezing. Receptors for trigeminal irritants are generally assumed to be located exclusively on free nerve endings within the nasal epithelium, requiring that trigeminal irritants diffuse through the junctional barrier at the epithelial surface to activate receptors. We find, in both rats and mice, an extensive population of chemosensory cells that reach the surface of the nasal epithelium and form synaptic contacts with trigeminal afferent nerve fibers. These chemosensory cells express T2R "bitter-taste" receptors and alpha-gustducin, a G protein involved in chemosensory transduction. Functional studies indicate that bitter substances applied to the nasal epithelium activate the trigeminal nerve and evoke changes in respiratory rate. By extending to the surface of the nasal epithelium, these chemosensory cells serve to expand the repertoire of compounds that can activate trigeminal protective reflexes. The trigeminal chemoreceptor cells are likely to be remnants of the phylogenetically ancient population of solitary chemoreceptor cells found in the epithelium of all anamniote aquatic vertebrates.