Fingerprinting taste buds: intermediate filaments and their implication for taste bud formation.

Intermediate filaments in taste organs of terrestrial (human and chick) as well as aquatic (Xenopus laevis) species were detected using immunohistochemistry and electron microscopy. During development, the potential importance of the interface between the taste bud primordium and non-gustatory adjacent tissues is evidenced by the distinct immunoreactivity of a subpopulation of taste bud cells for cytokeratins and vimentin. In human foetuses, the selective molecular marker for taste bud primordia, cytokeratin 20, is not detectable prior to the ingrowth of nerve fibres into the epithelium, which supports the hypothesis that nerve fibres are necessary for initiating taste bud development. Another intermediate filament protein, vimentin, occurs in derivatives of mesoderm, but usually not in epithelium. In humans, vimentin immunoreactivity is expressed mainly in border (marginal) epithelial cells of taste bud primordia, while in chick, vimentin expression occurs in most taste bud cells, whereas non-gustatory epithelium is vimentin immunonegative. Our chick data suggest a relationship between the degree of vimentin expression and taste bud cell proliferation especially during the perihatching period. It is suggested that surrounding epithelial cells (human) and mesenchymal cells (chick) may be contributing sources of developing taste buds. The dense perinuclear network of intermediate filaments especially in dark (i.e. non-sensory) taste disc cells of Xenopus indicates that vimentin filaments also might be associated with cells of non-gustatory function. These results indicate that the mechanisms of taste bud differentiation from source tissues may differ among vertebrates of different taxa.

Taste quality and intensity: lessons from the Morrison technique.

The nontoxic and nonshock Morrison operant technique was used to evaluate taste quality in rat and marmoset: response to a tastant test solution in pursuit of a pellet reward was dependent on making a choice between two bars that had been linked in discrimination training to qualitatively different stimulus pairs (NaCl versus either HCl, QHCl, or NH(4)Cl). The percentage distribution of bar-press responses to test stimuli showed: (1) stability of quality across 0.069-0.3 M NaCl, 0.003-0.1 M HCl, and 0.0001-0.003 M QHCl; (2) for LiCl, a quality change consistent with human reports of a "sour" to "salty" shift; (3) a suggestion that the "salty-like" quality of NH(4)Cl and NaCl are not perceptually equivalent; (4) NaNO(3) shares NaCl-like, QHCl-like, and NH(4)Cl-like components; (5) CaCl(2), KCl, and MgCl(2) share QHCl-like and NH(4)Cl-like components; and (6) responses to HCl and QHCl were not hedonically driven in the rat. Comparison of rank order correlations of single-unit firing rates to the distribution of bar-press responses for the same test stimulus concentration revealed that (7) no single level of the gustatory pathway exclusively accounts for the operant response distribution pattern to either simple or complex tastants, and (8) discriminations between tastants, one of which may be qualitatively complex, are not necessarily mediated only at levels proximal to the solitary nucleus. Thus, the Morrison discrimination technique effectively yields statements about gustatory quality without use of negative reinforcers and largely uninfluenced by tastant hedonics.

Organization of geniculate and trigeminal ganglion cells innervating single fungiform taste papillae: a study with tetramethylrhodamine dextran amine labeling.

Single gustatory nerve fibers branch and innervate several taste buds. In turn, individual taste buds may receive innervation from numerous gustatory nerve fibers. To evaluate the pattern of sensory innervation of fungiform papilla-bearing taste buds, we used iontophoretic fluorescent injection to retrogradely label the fibers that innervate single taste papillae in the hamster. For each animal, a single taste papilla was injected through the gemmal pore with 3.3% tetramethylrhodamine dextran amine. Fungiform papillae either at the tongue tip (0.5-1.5 mm from the tip) or more posteriorly (1.5-3.0 mm from the tip) were injected. After one to seven days survival, the geniculate and trigeminal ganglia and the tongue were sectioned and examined for labeled cells and fibers, respectively. Analysis of the number and topographic distribution of geniculate cells innervating single taste papillae, shows that: (i) 15 +/- 4 (S.D.) ganglion cells converge to innervate a single fungiform taste bud; (ii) more ganglion cells innervate anterior- (range: 13-35 cells) than posterior-lying buds (range: five to 12 cells), which, in part, may be related to bud volume (microm3); and (iii) ganglion somata innervating a single taste bud are scattered widely within the geniculate ganglion. Analysis of labeled fibers in the tongue demonstrated that two to eight taste buds located within 2 mm of the injected taste bud share collateral innervation with the injected taste bud. Since all buds with labeled fibers were located in close proximity (within a 2-mm radius), widely dispersed geniculate ganglion cells converge to innervate closely spaced fungiform taste buds. Trigeminal ganglion (mandibular division) cells were also labeled in every case and, as with the geniculate ganglion, a dispersed cell body location and collateralization pattern among papillae were observed. This study shows that iontophoresis of tetramethylrhodamine dextran amine, selectively applied to individual peripheral receptor end-organs, effectively locates sensory ganglion cells in two different ganglia that project to these sites. Moreover, the marker demonstrates collateral branches of sensory afferents associated with the labeled fibers and the nearby receptor areas innervated by these collaterals. The labeling of single or clusters of receptor cells, as well as identified sensory afferents, affords future possibilities for combining this technique with immunocytochemistry to establish the relationships of innervation patterns with neurotransmitters and neurotropic substances within identified cells.

Distribution of vimentin in the developing chick taste bud during the perihatching period.

The tissue environment within which taste bud cells develop has not been wholly elaborated. Previous studies of taste bud development in vertebrates, including the avian chick, have suggested that taste bud cells could arise from one, or several tissue sources (e.g. crest-mesenchyme, local ectoderm or endoderm). Thus, molecular markers which are present in gemmal as well as interfacing (peribud epithelium; mesenchyme-epithelium) regions, and their degree of expression during stages of taste bud development, are of special interest. The intermediate filament protein, vimentin, occurs in mesenchymal and mesodermally-derived (e.g. endothelial, fibroblast) cells as well as highly proliferating epithelium (e.g. tumors). The present study in chick gustatory tissue utilized antibodies against vimentin and the avidin-biotin-peroxidase technique to evaluate vimentin immunoreactivity (IR) within a timeframe which includes: 1) early stages of the taste bud primordium [embryonic days (E)17-E18)]; 2) the beginning of an accelerated bud cell proliferation at the time of initial, taste bud pore opening [around E19]; 3) attaining the adult complement of taste buds [around posthatch (H) day 1], and 4) completed organogenesis (H 17). During this time span, vimentin-IR was characterized in a region including and sometimes bridging taste bud and subepithelial connective tissue, whereas non-gustatory surrounding epithelium and salivary glands were vimentin-immuno-negative. Intragemmally, the proportion of vimentin-IR cells as related to total taste bud cells peaked at E19. These results indicate that vimentin expression, in part, is related to the onset of taste bud cell proliferation and suggest that mesenchyme could be one source of taste bud cells. Secondly, fibronectin, an extracellular matrix component of the epithelial basement membrane interface with mesenchyme, was expressed at or near the apical surfaces of taste bud cells projecting into the bud lumen, and in the basal gemmal region suggesting the possible role of fibronectin as a chemotactic anchor for differentiating and migrating taste bud receptor cells. Lastly, neuron-specific enolase-IR indicates that axonal varicosities are already present intragemmally at E17-E18, that is, during the incipient period of identifiable taste bud primordia.

Identified taste bud cell proliferation in the perihatching chick.

Developing taste buds in the anterior mandibular floor of perihatching chicks were studied by high voltage electron microscopic autoradiography in order to identify proliferating gemmal cell types. Montaged profiles of 29 taste buds in five cases euthanized between embryonic day 21 and posthatching day 2 were analyzed after a single [3H]thymidine injection administered on embryonic day 16, 17 or 18. Results showed that dark cells comprised 55% of identified (n = 900 cells) and 62% of labeled (n = 568 cells) gemmal cells as compared with light, intermediate, basal or perigemmal bud cells. Dark cells had both a greater (P < 0.05) number of labeled cells and a greater amount of label (grains/nucleus) than the other four bud cell types, irrespective of injection day. The nuclear area (micron 2) of dark cells was not significantly larger (P > 0.05) than that of the other gemmal cell types and therefore cannot account for the greater amount for label in the dark cells. Interestingly, only dark cells showed a positive correlation (P < 0.003) between amount of label and nuclear area. Results suggest that, during the perihatching period of robust cell proliferation, dividing dark cells may give rise primarily, but not exclusively, to dark cell progeny.

Neural cell adhesion molecule, neuron-specific enolase and calcitonin gene-related peptide immunoreactivity in hamster taste buds after chorda tympani/lingual nerve denervation.

Hamster fungiform papilla taste buds persist in an atrophic form following sensory denervation. While atrophic and innervated taste buds are morphologically similar, it is not known whether their gemmal cells have similar molecular characteristics. Three neurochemicals, neural cell adhesion molecule, neuron-specific enolase, and calcitonin gene-related peptide have been implicated in trophic phenomena, synaptogenesis and cell recognition in neurons and sensory neuroepithelia. The present study uses immunocytochemical localization of these molecular markers to characterize normal and denervated fungiform taste buds following unilateral chorda tympani/lingual nerve denervation in hamsters. In normal taste buds, immunoreactivity to neural cell adhesion molecule, neuron-specific enolase, and calcitonin gene-related peptide was present in a group of cells located centrally in the bud as well as in fungiform nerve fibres and endings. After denervation, gemmal cell immunoreactivity to all three markers was reduced and often confined to a single or a few bud cell(s). Also, fibre staining was absent except for sparse calcitonin gene-related peptide-immunoreactive fibres associated with blood vessels and within the fungiform papillae. These remaining fibres may be autonomic or somatomotor in origin. These results indicate that sensory denervation of hamster taste buds reduces, but does not wholly eliminate the immunoreactivity of surviving gemmal cells to neural cell adhesion molecule, neuron-specific enolase, and calcitonin gene-related peptide. While the number of taste bud cells expressing the markers appears to be nerve-dependent, immunoreactivity in sensory-denervated bud cells of hamster may reflect the influence of local tissue factors.

Pontine reticular formation is involved in catalepsy produced by cholinergic drugs.

Bilateral kainic acid lesions of the ventro-medial (VM) thalamic nucleus of rats which greatly reduced the catalepsy produced by haloperidol (2 mg/kg i.p.) not only did not reduce, but even enhanced, the cataleptogenic effect of eserine (1 mg/kg i.p.) and arecoline (30 mg/kg i.p.). This finding is in accord with former conclusions that catalepsy produced by cholinergic drugs does not depend on striatal mechanisms. In rats with kainic acid lesions of the VM thalamic nucleus, and similarly in intact, non-lesioned rats, systemic administration of eserine and arecoline potentiated the catalepsy produced by microinjections of carbachol (2 microg) into the pontine reticular formation (PRF). Atropine microinjected bilaterally into the PRF attenuated the cataleptogenic effect of eserine and arecoline i.p. We suggest that the PRF is a site at which systemically given cholinergic drugs act to produce catalepsy.

Taste bud cell generation in the perihatching chick.

Chick taste bud primordia initially appear in late gestation on embryonic day 17 (E17), 4 days before hatching. To track DNA synthesis and subsequent taste bud cell proliferation between E17 and the second day post-hatching (H2), single 25 muCi injections of tritiated thymidine (specific activity = 72.5 Ci/mmol) were administered in ovo during E15, E16, E17 or E18. Anterior mandibular oral epithelium was processed for light microscopic autoradiography. Sections through each taste bud's center were analysed for label (> or = 6 silver grains/gemmal cell nucleus), and bud diameter. Results indicated a major part of gemmal cell DNA synthesis does not occur until after E19 irrespective of the day of thymidine injection, suggesting postmitotic or quiescent (decycled) cells assemble to form the early bud primordium (E17-19) based on local tissue interactions. All buds examined from E20-H2 contained labelled cells. The day of injection was important since 5-day survival cases after E16 injection yielded about 25% the number of labelled cells/bud as compared with equivalent survival cases following E17-18 injections. These results are discussed with respect to parallel changes in bud shape and increasing bud diameter, and cell proliferation in possible extra- and intragemmal sources of bud cells.

Ontogenesis and taste bud cell turnover in the chicken. I. Gemmal cell renewal in the hatchling.

Taste bud cell turnover rate was examined in oral epithelium of the precocial chick, which at hatching contains the adult complement of taste buds. Forty newly hatched chicks received single or double pulse injections of tritiated thymidine (specific activity was 6.7 Curies/millimole; dosage was 0.5 microCuries/g body weight, intraperitoneally). Anterior mandibular epithelium was processed for light microscopic autoradiography at 2 and 16 hours, as well as 1, 2, 3, 4, 6, 8, 10, 12, 14, 16, 18, and 20 days after the initial pulse. In a coded and randomized procedure, the section (7 microns) through the bud's center was selected for counting > or = 6 silver grains over round-clear and gracile-dense gemmal cell nuclei. The mean number of labelled cells/bud varied significantly (P < or = 0.01) during the first four posthatch days, yielding the fastest gemmal cell turnover rates (3.4-4.4 days) yet reported in vertebrates. Average bud diameter also significantly changed during the first four posthatch days, and was reflected in shifts of the distribution of 40-69 microns and > or = 70 microns diameter buds. Both an increase in labelled bud cells and bud diameter during the first two posthatch days may reflect high proliferation rates in initially maturing buds. Subsequent decrease in bud diameter between 2 and 3 days postinjection may indicate splitting of large-diameter (> or = 70 microns) buds and/or normal bud cell death due to failure of sensory afferentation. Bud-splitting alone, however, cannot account for significant decreases in bud cell label which did not occur before 4-6 days postinjection.

Aspects of vertebrate gustatory phylogeny: morphology and turnover of chick taste bud cells.

The taste bud is a receptor form observed across vertebrates. The present report compares chick taste buds to those of other vertebrates using light and electron microscopy. Unlike mammals, but common to many modern avians, the dorsal surface of chick anterior tongue lacks taste papillae and taste buds. Ultrastructurally, chick buds located in the anterior floor of the mouth (as in some reptiles and amphibians) and palate contain dark, intermediate, light, and basal cell types. Dark, intermediate, and light cells extend microvilli into intragemmal lumina and pores communicating with the oral cavity. As specialized features, dark cell apices lack dense granules and exhibit short microvilli relative to light and intermediate cells. Dark cell cytoplasmic fingers envelop intragemmal nerve fibers and cells as in other species, and sometimes contain abundant clear vesicles. Nerve profile expansions often are located adjacent to dark, intermediate, and light cell nuclei. Classical afferent synaptic contacts are rarely observed. Taste cell turnover is suggested by mitotic and degenerating figures in chick buds. In addition, tritiated thymidine injected into hatchlings, whose anterior mandibular oral taste bud population approximates that in adults, reveals a turnover rate of about 4.5 days. This is about half that observed in altricial mammals, reflecting a species difference or developmental factor in precocial avians. It is concluded that chick taste buds exhibit morphologic features common to other vertebrate buds with specializations reflecting the influences of niche, glandular relations, and/or age.

Ultrastructure of palatal taste buds in the perihatching chick.

Palatal taste buds of perihatching chicks were examined by electron microscopy. Four intragemmal cell types were characterized. 1) Light: with voluminous, electron-lucent cytoplasm containing scattered free ribosomes, rough and smooth endoplasmic reticulum, plump mitochondria, sparse perinuclear filaments, occasional Golgi bodies, and numerous clear and dense-cored vesicles. Clear vesicles sometimes aggregate in a presynaptic-like configuration apposed to an axonal profile. These cells contained large, spherical, uniformly granular nuclei with one nucleolus. 2) Dark: with dense cytoplasm containing filamentous bundles surrounding the nucleus, occasional clear vesicles, centrioles, rough endoplasmic reticulum, and compact mitochrondria. The apical cytoplasm noticeably lacks dense secretory granules. Irregular to lobulated nuclei are densely granular, and contain scattered clumps of chromatin, adhering especially to the inner leaflet of the nuclear membrane, and at least one nucleolus. Cytoplasmic extensions of dark cells envelop other intragemmal cell types and nerve fibers. Light and dark cells project microvilli into the taste pore. 3) Intermediate: contain gradations of features of light and dark cells. 4) Basal: darker than the other intragemmal cell types and confined to the ventral bud region. Putative afferent synapses in relation to light cells, and axo-axonal contacts are described. While the appearance of axo-axonal contacts may be a transient developmental event, other bud features are consonant with observations in adult chickens and suggest that the peripheral gustatory apparatus is mature at hatching in this precocial avian species.

Dopaminergic drugs influence the intensity of catalepsy induced by microinjections of carbachol into the reticular formation.

Carbachol microinjections into the mesencephalic and pontine reticular formation in rats induced intense and long-lasting catalepsy. Systemically administered haloperidol potentiated, while apomorphine and L-DOPA reduced the cataleptogenic effect of carbachol. These results indicate the existence of functional relations between the cholinergic cataleptogenic mechanism in the reticular formation and the dopaminergic system. They are interpreted in the light of known anatomical ascending and descending interconnections between the reticular formation and basal ganglia.

Long-term effects of gustatory neurectomy on fungiform papillae in the young rat.

Recent evidence from mature hamster fungiform papillae indicates that following denervation taste buds are present from 21 to 330 days in the absence of discernible intragemmal nerve fibers. In contrast, most prior taste bud degeneration studies focused on shorter survival times. The present inquiry in young rats examined the issue of postneurectomy buds, in which regeneration of the resected chorda tympani or facial nerves was prevented and anterior tongue tissue examined over a range of relatively long survival times (30-90 days). Conditions for observing potential taste buds used three histologic stains and a definition of the taste bud not necessarily requiring pore identification. In each case, serial section examination of the anterior-most 2-3 mm of lingual epithelium revealed 29-56 bud-containing fungiform papillae on the unoperated side. In contrast, ipsilateral to the neurectomy, only zero-7 medially-placed, mature-looking buds were observed per case, as well as zero-3 more laterally situated fungiform papillae containing small clusters of cells in basal epithelium that lacked the vertical organization and cytoplasmic staining intensity of mature taste buds. These cell aggregates were distributed evenly across survival time and stain used. Therefore, in young rats following gustatory neurectomy, longer survival times, per se, would not appear to be a prerequisite for sustaining fungiform taste buds. The appearance of "midline" buds postsurgery may be attributed to either normal contralateral or a net bilateral innervation, and/or ipsilateral denervation and bud loss inducing neural sprouting from the contralateral side.

Central distribution and efferent origins of facial nerve branches in the chicken.

The central afferent distribution and the origins of visceromotor and somatomotor components of greater superficial petrosal (GSP), chorda tympani (CT), and hyomandibular (Hy) branches of the facial nerve were analyzed by the transganglionic and retrograde transport of horseradish peroxidase. Labelled sensory afferent fascicles of the GSP collected dorsomedial to the spinal trigeminal complex and passed caudally as the tractus solitarius. Fine-grain terminal fields were evident in ipsilateral n. sensorius n. facialis, nuclei ventrolateralis anterior (Vla), centralis anterior and presulcalis anterior of the solitary complex, and n. glossopharyngei et n. motorius dorsalis nervi vagi (n. IX-X). Minor contralateral projections to these nuclei occasionally were observed. CT sensory afferents exhibited similar ipsilateral projections, and specifically, the projection to Vla was qualitatively sparser than that from the GSP. Small multipolar and spindle-shaped somata of visceromotor n. salivatorius were labelled retrogradely from the GSP and CT. Large, oval somatomotor cells of the three subdivisions of the facial motor nucleus were labelled retrogradely from Hy, with no evidence of sensory afferent or visceromotor connections.

Taste bud development in chickens (Gallus gallus domesticus).

Oral epithelium in the anterior mandibular glands region was examined in embryonic, hatchling, and mature chickens to establish the timing of morphologic events during taste bud ontogeny. Hematoxylin-and-eosin-stained sections (10 microns) from 27 Anak (broiler breed) chickens were examined serially, and buds were quantified at 16-20 days of incubation (E) and, posthatch days 1 and 50-60. Taste buds were first recognized at the beginning of E17 as small clusters of cells in the basal epithelium. Only spherical-shaped buds were observed on E17 and E18, and these spherical clusters never penetrated to the surface of the stratified epithelial layer. E19 marked a transitional stage when mature bud features began to emerge: the buds assumed a more elongate shape, several kinds of cells comprising the bud were distinguishable and the first taste pores were observed. During the ensuing embryonic days, buds continued to elongate commensurate with the deepening oral epithelium and by hatching virtually all buds opened to the oral cavity. No marked morphological changes in taste bud structure were observed on the day of hatching and at 50-60 days posthatching. Taste bud numbers increased dramatically during E17 and E18, peaked on E19, and remained relatively constant thereafter. It is concluded that the morphological sequence of taste bud development in chickens is similar to that in mammals. The timing of bud ontogeny, though initiated only during the third trimester in ovo, essentially is completed by hatching, thus providing the precocial hatchling with the sensory apparatus essential for gustatory experience.

Chorda tympani innervation of anterior mandibular taste buds in the chicken (Gallus gallus domesticus).

While the mammalian chorda tympani innervates taste buds on the anterior two-thirds of the tongue, the chorda tympani of chickens does not enter the tongue, but rather is reported to supply the oral epithelium of the lower beak subjacent to the tongue. This study in the chicken investigated whether the integrity of taste buds in the lower beak is normally dependent upon innervation by the chorda tympani. Following unilateral ligation and removal of a large section of the chorda tympani, animals were sacrificed at 11, 14, and 21 days postoperatively. Oral tissue between the lingual frenulum and beak tip was serially examined, and the presence of each bud was recorded, noting the point at which the bud opened into the oral cavity. No buds were observed on the operated side in any of the cases, while the average bud count on the unoperated side was 33 +/- 10 (SD). On the unoperated side, taste buds were generally associated with anterior mandibular salivary gland ducts that reached surface epithelium and opened into the oral cavity. On the operated side, the cellular organization adjacent to gland ducts and in duct-free epithelium appeared as in control (i.e., bud-free) epithelium. The number of salivary gland duct openings into the oral cavity was equivalent on the operated and control sides. It is concluded that the chorda tympani of chickens innervates taste buds in the anterior lower beak epithelium and that it functions to maintain the structural integrity of these buds.

Dual efferent projections of the trigeminal principal sensory nucleus to the thalamic ventroposteromedial nucleus in the squirrel monkey.

Anterograde degeneration methods demonstrated two efferent components from the trigeminal principal sensory nucleus (PrV) to the thalamic ventroposteromedial nucleus (VPM) in the squirrel monkey: fibers from the dorsal PrV coursed within the central tegmental tract and terminated in a dorsoventromedial strip of the ipsilateral VPM; fibers from the ventral PrV mainly decussated caudal to the interpeduncular nucleus and terminated in the contralateral VPM exclusive of the sector receiving the dorsal PrV component, contralaterally. Adjacent Nissl sections showed an apparent increase in glial profiles accompanying an intense somal staining among the deafferented neuronal population in the VPM, coextensive with those regions in the VPM exhibiting terminal field degeneration.

Central afferent connections and origin of efferent projections of the facial nerve in the chicken (Gallus gallus domesticus).

The central afferent connections and origin of efferent projections of the facial nerve in the adult domestic chicken were studied by anterograde and retrograde transport of horseradish peroxidase from the geniculate ganglion. Ipsilateral afferent projections were traced caudal to the level of entrance of the facial nerve and into tractus solitarius (TS), located dorsomedial to the spinal trigeminal nuclear complex. At several rostrocaudal levels in the medulla, fibers exited from TS and terminated in n. sensorius N. facialis (SVII), and nn. ventrolateralis anterior (Vla) and intermedius anterior (Ia) solitarii. Some axons were followed to n. presulcalis anterior (Pas) solitarii. A separate component terminated in subnucleus interpolaris (ip) of n. descendens nervi trigemini or its medially adjacent reticular formation either by exiting from TS or coursing caudally through the trigeminal complex from entering facial rootlets. Another diffuse component of facial axons ascended dorsally and rostrally from the level of entrance of the facial nerve; these projections dissipated in the pons--some on the dorsomedial border of n. principalis N. trigemini (PrV). Ipsilateral efferent projections were traced through the main genu of the facial nerve to retrogradely labelled somata of pars dorsalis (FMd), pars intermedia (FMi), and pars ventralis (FMv) of n. motorius nervi facialis. A separate group of smaller, multipolar, and spindle-shaped cells (8-25 microns) wedged between n. olivaris superioris (OS) and the caudal end of FMv, rostrally, and extending caudally in ventrolateral medulla were labelled. These small cells contrast with the larger (21-45 microns), oval, round, and multipolar somata of FMv and may correspond, in part, to a parasympathetic n. salivatorius (Sal).

Number and distribution of taste buds in the oral cavity of hatchling chicks.

The location and number of taste buds were mapped in palatal epithelia of one-day old chicks and bud widths measured. Bud counts additionally were recorded for the tongue, and floor of the lower beak. An average of 316 taste buds was observed in the oral cavity of which 69%, 29% and 2% were distributed across oral epithelium in the upper beak (palate), lower beak and posteroventrolateral region of the anterior tongue, respectively. In each oral region, salivary gland ducts lying adjacent as well as gland ductules penetrating through the buds were prevalent. This relation may provide the bio-fluid milieu for receptor stimulation during feeding. Widths of palatal buds were bimodally distributed, peaking at diameters between 40-49 and 60-69 microns. The taste bud-rich oral epithelium in these one-day old chicks is consonant with their precocial nature. The topographic distribution of taste buds appears to be in register with those regions of epithelium contacted by food which is transported anteroposteriorly through the oral cavity by the chicken's prehensile tongue.

Thoracic dorsal funicular lesions affect the bouton patterns on, and diameters of, layer VB pyramidal cell somata in rat hindlimb cortex.

The effect of spinal dorsal funicular lesions (T 12) upon the frequency of boutons on, and diameters of the somata of pyramidal cells in layer VB of hindlimb cortex was studied. Adult rats sustained bilateral damage to either the dorsal column (DC, n = 10) alone or DC combined with the corticospinal tract (CS) (DC + CS, n = 34) and were utilized 1, 2, 3, 7, 14, 30, 45, 60, 90, or 120 days postoperatively (DPO). Neurons randomly sampled from 44 lesioned and 13 unoperated cases were analyzed for the number of silver-impregnated boutons (Rasmussen method) on the circumference of the soma as well as diameters of the soma, nucleus, and nucleolus. Analyses of variance comparing across lesioned and normal groups were significant for bouton counts on the soma (P less than 0.01), and diameters (long axis) of somata (P less than 0.01) and their nuclei (P less than 0.05). Both lesioned groups exhibited significant decreases from normal for these latter three parameters. With respect to survival time for the DC + CS-lesioned animals we noted the following: (1) Bouton counts on the soma significantly decreased below normal between 1 and 60 DPO; this decrease was most dramatic during the first three days postlesion. (2) Somal diameter (long axis) significantly decreased below normal between 2 and 120 DPO (except at 14 and 90 DPO). (3) Nuclear diameter (long axis) significantly decreased below normal only at 90 DPO. (4) Bouton counts on somata of neurons in layers VB and IV [Ganchrow and Bernstein, 1981] of hindlimb cortex correlated negatively and significantly across 120 postlesion days. The rapid shrinkage and reduced afferentation of layer VB somata during the first week following DC + CS lesions suggest initial, retrograde reactions to CS axotomy. Since bouton counts on layer VB somata were significantly less (P less than 0.05) in DC- than DC + CS-lesioned rats, it is hypothesized that CS axotomy regulated a set-point for increased afferentation which was maintained on the shrunken somata between 7 and 120 DPO.