Some aspects of the relationship between the structure of a bitter diketopiperazine and its receptor.
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Brain-derived neurotrophic factor (BDNF), one of the members of the nerve growth factor family of neurotrophins, is expressed in developing gustatory papillae and is thought to be the neurotrophin that supports gustatory innervation during development. BDNF expression does not cease after development but continues in some taste cells of adult mice. To determine which types of taste cells produce BDNF, we undertook an immunohistochemical study of taste cells in BDNF(LacZ) gene targeted "knock-in" adult mice. In these mice, beta-galactosidase (beta-gal) immunoreactivity is an indicator of cells that produce BDNF transcripts. In the tongues of adult BDNF(LacZ) mice, beta-gal (BDNF) is present in long slender taste cells, as well as pyriform taste cells. Bromodeoxyuridine labeling experiments in BDNF(LacZ) mice indicate that BDNF is not present in taste cells that are younger than 3 days postmitotic. BDNF mainly colocalizes with markers of type II and type III taste cells: ubiquitin carboxyl terminal hydrolase (PGP 9.5), serotonin (5-HT), neural cell adhesion molecule (N-CAM), synaptic associated protein of 25 kDa (SNAP-25), and to a lesser extent with alpha-gustducin. beta-Gal immunoreactivity is not associated with blood group H antigen, a marker of type I taste cells. We conclude that BDNF is absent from basal cells and type I (blood group H antigen immunoreactive) taste cells but is present in differentiated type II and type III taste cells. The presence of SNAP-25 in BDNF-expressing cells suggests a role for BDNF in synaptic formation and transmission.
Early dietary sodium restriction has profound influences on the organization of the gustatory brainstem. However, the anatomical relationships among multiple gustatory nerve inputs have not been examined. Through the use of triple-fluorescence labeling and confocal laser microscopy, terminal fields of the greater superficial petrosal (GSP), chorda tympani (CT), and glossopharyngeal (IX) nerves were visualized concurrently in the nucleus of the solitary tract (NTS) of developmentally sodium-restricted and control rats. Dietary sodium restriction during pre- and postnatal development resulted in a twofold increase in the volume of both the CT and the IX nerve terminal fields but did not affect the volume of the GSP terminal field. In controls, these nerve terminal fields overlapped considerably. The dietary manipulation significantly increased the overlapping zones among terminal fields, resulting in an extension of CT and IX fields past their normal boundaries. The differences in terminal field volumes were exaggerated when expressed relative to the respective NTS volumes. Furthermore, increased terminal field volumes could not be attributed to an increase in the number of afferents because ganglion cell counts did not differ between groups. Taken together, selective increases in terminal field volume and ensuing overlap among terminal fields suggest an increased convergence of these gustatory nerve terminals onto neurons in the NTS. The genesis of such convergence is likely related to disruption of cellular and molecular mechanisms during the development of individual terminal fields, the consequences of which have implications for corresponding functional and behavioral alterations.
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The second order projections of the gustatory system in catfish were examined using both retrograde (HRP) and anterograde (degeneration and autoradiographic) hodological methods. Golgi-stained material was used to demonstrate the different cell types in the primary gustatory sensory area, the facial lobe. Efferents from the facial lobe gather into ascending and descending secondary gustatory tracts. The descending tract terminates largely in the medial funicular nucleus and the commissural nucleus of Cajal in the region of the obex. A small portion of the descending tract continues caudally to terminate in the dorsal horn of the spinal cord. The ascending secondary gustatory tract terminates mostly in the ipsilateral superior secondary gustatory nucleus in the isthmic region. A small portion of the ascending tract continues rostrally to terminate in the posterior thalamic nucleus and in the region of the nucleus lobo-bulbaris. Sparser contralateral projections are also seen in the posterior thalamus and isthmic gustatory regions. Three cell types can be discerned in the facial lobe: small, medium and large. The small cells are intrinsic neurons, the medium cells project to the isthmic gustatory nucleus, and the large cells send fibers to the other terminal areas described above, as summarized in figure 19.
The central fiber connections of the gustatory system (VII, IX, and X nerves) in th crucian carp were examined by the Fink-Heimer method and its modification. The sensory and recurrence roots of the VII enter th brainstem separately and terminate in teh ipsilateral half of the facial lobe (L-VII). Afferent fibers of the IX terminate in the glossopharyngeal lobe (L-IX). Most afferent fibers of the X terminate in the sensory layer of the vagal lobe (L-X), in which degenerating terminals occur in some laminae. Some vagal afferents project bilaterally to the commissural nucleus of Cajal. The cutaneous component of the X projects to the nucleus of the spinal trigeminal tract (SpV) and the medial funicular nucleus (nFM). Ascending secondary fibers from the L-VII project bilaterally to the secondary gustatory nucleus (nGS) in the isthmus region. Descending secondary fibers from the L-VII turn caudally in the SpV. These fibers terminate mostly in the nucleus of the SpV and sparsely in the nFM. The L-IX and L-X give rise to the long and short secondary paths. The long path projects as the ascending secondary tract to the ipsilateral nGs. The short path includes secondary fibers projecting to the motor layer of the L-X and the medullary reticular formation. Teritary gustatory fibers arisig in the nGs project ipsilaterally to two diencephalic nuclei; the nucleus glomerulosus and the nucleus diffusus lobi inferioris.
To learn whether neurophysiological taste responses change during structural development of the gustatory system, we recorded from the chorda tympani nerve in rats aged 7 to 92 days after birth. Chemical stimuli applied to the anterior tongue included four monochloride salts, two acids, sucrose, and urea. Responses to all chemicals were obtained as early as 7 days postnatally. Developmental changes in salt, acid, and sucrose responses were observed. Relative to NaCl and LiCl, NH4Cl and KCl gradually decrease in effectiveness as taste stimuli; or, relative to NH4Cl and KCl, NaCl and LiCl become more effective stimuli. These changes are similar to those observed prenatally and postnatally in sheep. Also, relative to NaCl, citric acid, hydrochloric acid, and sucrose become less effective stimuli; or, NaC1 becomes more effective as a stimulus, relative to these acids and sucrose. The period of most rapid functional change overlaps a period of rapid structural change. It seems most reasonable to hypothesize that the altering taste responses reflect developmental changes in receptor membrane composition. Since the taste system is not programmed to respond in a mature manner from the moment function begins, there is ample opportunity for changing taste experience to influence the developing taste system.
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Fetal macaque chemosensory corpuscles during the last part of gestation contained chemosensory, sustentacular, and undifferentiated basal cells. Sustentacular cells had apical secretory granules and no specialized contacts with axons. Chemosensory cells contained basal collections of 80-100 nm dense core granules, and specialized axonal contacts of three types--afferent synapses, efferent synapses, and subsurface cisternae. Afferent synapses were commonly present on electron opaque cells with many 80-100-nm granules, typical 40-60-nm synaptic vesicles, and a few cisternae of smooth or granular endoplasmic reticulum. Cells with subsurface cisternae and/or efferent synapses were usually electron lucent, lacked vesicles and granules, and contained numerous intracytoplasmic cisternal elements. A continuum of intermediate forms was observed. It is postulated that transition of synaptic arrays accompanies the maturation of individual chemosensory cells.
Fungiform papillae contained well-developed chemosensory corpuscles in macaque monkey fetuses with crown-rump lengths of 5.0-9.0 cm. These fetuses corresponded to stages from the last part of the first and beginning of the second trimester of gestation. Chemosensory cells extended from the epithelial basal lamina to the taste pore anlage at the apex of the corpuscle and had typical afferent synaptic contacts with presumptive gustatory axons. Apical secretory granules, the distinctive cytologic feature of sustentacular cells, were absent. Cells with cytologic characteristics intermediate between chemosensory cells and basal extragemmal cells were considered to be differentiating chemosensory (DC) cells and appeared to arise from existing basal cells as well as undifferentiated postmitotic cells. Although extragemmal cells were extensively coupled by gap junctions which are apparent in electron micrographs of thin sections, similar junctions were absent from chemosensory and DC cells. While having minimal structural requirements for transduction and transmission of sapid stimuli (i.e., an intraoral surface and afferent synaptic contacts) chemosensory cells lacked efferent synapses and subsurface cisternae, which can be observed at later stages of gestation. Axoaxonic synapses, present between gustatory axons in the intragemmal and extragemmal epithelium and in the subgemmal connective tissue, were observed with greater frequency than at later stages of gestation. We conclude that the differentiation of chemosensory cells precedes the differentiation of sustentacular cells by several weeks. This fact lends support to the concept that chemosensory cells are a unique identifiable cells type in all chemosensory corpuscles.
The ultrastructure and identification of WGA-HRP-labeled sensory receptors in the rat incisive papilla (the most anterior part of hard palate) were studied using semiserial thin sections. Various sensory receptors were organized according to three locations: dome region (ventral), chemosensory corpuscle region (medial to orifice of incisive canal), and lateral labium (apposing the incisive canal). In the dome region, the sensory receptors were localized in three sensory zones that were associated with surface ridges (one medial and two lateral). In each of these zones, intraepithelial receptor axons and Merkel receptors occurred in the epithelium, while simple unencapsulated corpuscles, glomerular-Meissner corpuscles, and incisive (encapsulated) corpuscles occurred in the lamina propria. In the chemosensory corpuscle region, chemosensory corpuscles and intraepithelial receptor axons were located in the epithelium, and incisive corpuscles were present in the lamina propria. In the lateral labium, only intraepithelial receptor axons were prominent. In all these sensory receptors, the preterminal axons and axon terminals were labeled with the tracer protein. In addition, some nonneuronal cells closely associated with the axon terminals were selectively labeled, e.g., terminal Schwann cells, lamellar Schwann cells, Merkel cells, corpuscular basal cells and chemosensory cells. Other adjacent cells were not labeled, e.g., unspecialized epithelial cells, capsular cells, corpuscular sustentacular cells, and fibroblasts. In both labeled axons and cells, WGA-HRP was incorporated into vesicles, tubules, and vacuolar organelles. The specific intercellular transfer of tracer protein may indicate trophic interactions between axon terminals and support cells in sensory receptors. The specific organization of multiple sensory receptors in the rat incisive papilla may provide a useful alternative system for studying somatosensory physiology.
Human infants less than 4 months of age ingested water and moderate concentrations of sodium chloride solution (0.10-0.20 M) in equal amounts. In contrast, infants 4-24 months of age exhibited heightened acceptance of saline solution relative to water. Such a developmental change in salt acceptability is consistent with a hypothesis of postnatal maturation of peripheral and/or central structures underlying this response. Children 31-60 months of age tended to reject saline solution relative to water. This change may in part reflect the effects of experience.
Intraoral lemon infusion elicits facial wiping behavior in fetal rats on Days 20 and 21 of gestation. On Day 19, fetuses tested in a warm saline bath respond to infusion but do not express the wiping response. In contrast, fetuses tested within the amniotic sac exhibit wiping. Fetal movements after infusion were analyzed on Days 19 and 20 in both the bath and amnion environments. Facial wiping is assembled from simpler components in both real time (during the 15 s after infusion) and developmental time (over Days 19-20 of gestation). In real time, the wiping response occurs following a brief period of fetal activity comprising head and forelimb movements. After the wiping bout is initiated, strokes by left and right forelimbs increase in duration and become synchronized. In developmental time, components of the wiping response (forelimb strokes) are consistently expressed on Day 19, but other components (head stabilization) are lacking when fetuses are tested in bath. The amniotic sac appears to provide scaffolding that reduces head movement and promotes paw-face contact, thus facilitating facial wiping.
Hydrational state has been demonstrated to influence intake of various solutions in young rat pups. For instance, both cellular and extracellular dehydration produce an enhancement of intake in pups tested at 6 days of age. However, the behavioral mechanisms that result in increased intake following manipulations of hydrational state have been less extensively studied. The impact of hydrational state on behavioral responsiveness in young rat pups was examined by assessing the pattern of responding to a series of repeated oral infusions of diet. Pups were tested at 6, 12, or 18 days of age following either acute cellular dehydration produced by injection of 1 M NaCl or acute extracellular dehydration produced by injection of 10% polyethylene glycol (PEG). Oral responsiveness to a series of 30 brief infusions of one of four taste solutions (water, 10% sucrose, 0.135 M NaCl, or 1 M NaCl) was measured. Each infusion lasted 3 s and there was 1 min between infusions. The pattern of oral responding to solutions was affected by the developmental age of the pup, the hydrational state of the pup, and the solution offered, with the largest effects of dehydration observed in the youngest animals. In all conditions except one, pups habituated to repeated infusions. The exception was the failure of extracellularly dehydrated 6-day-old pups to display habituation to oral infusions of sucrose. These results suggest that, although intake is enhanced by both cellular and extracellular dehydration in very young pups, the behavioral changes responsible for the enhancement of intake after cellular dehydration are different from the behavioral changes resulting from extracellular dehydration. This dissociation of behavioral effects of dehydration in young pups demonstrates that intake measures alone may obscure subtle differences in behavior and argues for the utility of dissection of behavioral components in understanding the neural and physiological control of behavior.
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HRP histochemistry and microelectrode mapping were combined to study the sizes, shapes, and orientations of neuronal cell bodies and dendrites located at sites of taste-elicited single unit activity in the nucleus of the solitary tract (NST). Cells responding to sapid stimulation of the anterior tongue were extracellularly recorded using micropipettes containing HRP. Iontophoretic injection of the marker at the recording sites resulted in small (50-200 microns diameter) opaque zones bordered by a small number (2-15) of neurons with Golgi-like filling of their cell bodies, dendrites, and to some extent, their axons. The cell bodies were near (50-250 microns) the injection sites into which they sent labelled dendrites. Two broad categories of neurons were typically filled. Elongate cells had oval- to spindle-shaped cell bodies oriented mediolaterally. Two primary dendrites extended 100-300 microns from the cell body, one medially and one laterally, and branched within a cylindrical dendritic field oriented mediolaterally. A minority of the HRP-filled elongate cells had unusually long rostrally or caudally directed dendritic branches. Stellate cells had oval, round, triangular, or polygonal cell bodies and 3-5 primary dendrites coursing 200-300 microns in all directions and branching as unoriented, spheroidal fields. A minority of stellate cells had relatively unbranched wavy dendrites, resembling tentacles, while others had unusually small cell bodies (10-15 microns diameter), small dendrites, and locally arborizing axons. Of 151 labelled cells, all but 12 were remarkably confined to the rostral NST. Nearly 90% were concentrated in the rostral central cytoarchitectonic subdivision, where stellate cells predominated, or in the rostral lateral subdivision, where elongate cells predominated. These morphological types of neurons, filled at neurophysiological recording sites, are compared with cell types identified in previous light and electron microscopic studies of the cytoarchitecture, connections, and synaptic organization of the gustatory NST.
A clear understanding of the role of different papillae in the pig may permit the development of a more palatable diet, thus, better utilizing anatomical structures and morphological characteristics to enhance health and productivity in this domestic species. The study used ten healthy market-size, mixed-breed pigs to properly describe the surface structure of the tongue papillae using standard scanning electron microscopic techniques. Two types of sharp and blunt filiform papillae were identified. Fungiform papillae, located on the lateral side of the tongue, were circular and large with surface taste pores. The majority of oval papillae on the dorsal surface of the tongue contained multidirectional surface grooves and lacked taste pores. The foliate papillae, located principally on the caudal third and lateral part of the tongue, were comprised of four to five leaves and they had additional pseudopapillae. The two vallate papillae, found in the caudal third and dorsum of the tongue, had a well-defined vallum and pseudopapillae on their surface. At the root of the tongue, two types of conical papillae with or without apex were observed. The entire area covered by conical papillae was deeply occupied by lingual tonsils. The lingual tonsils were scattered with varying sized lymph nodules in the propria mucosa of the conical papillae deep to epithelium. Functionally, the filiform and conical papillae appeared to have a mechanical role whereas fungiform, foliate, and vallate papillae apparently perform a gustatory role in the pig. The tonsillar location at the root of the tongue provides an ideal experimental model for immunohistological and immunological studies on the lymphoid tissue in man.