Recurrent facial taste neurons of sea catfish Plotosus japonicus: morphology and organization in the ganglion.

This study investigated the morphology of the recurrent facial taste neurons and their organization in the recurrent ganglion of the sea catfish Plotosus japonicus. The recurrent ganglion is independent of the anterior ganglion, which consists of trigeminal, facial and anterior lateral line neurons that send peripheral fibres to the head region. The recurrent taste neurons are round or oval and bipolar, with thick peripheral and thin central fibres, and completely wrapped by membranous layers of satellite cells. Two peripheral nerve branches coursing to the trunk or pectoral fin originate from the recurrent ganglion. The results presented here show that the trunk and pectoral-fin neurons are independently distributed to form various sizes of groups, and the groups are intermingled throughout the ganglion. No distinct topographical relationship of the two nerve branches occurs in the ganglion. Centrally, the trunk and pectoral-fin branches project somatotopically in the anterolateral and intermediate medial regions of the trunk tail lobule of the facial lobe, respectively.

Early detection of radiation-induced structural changes in rat trabecular bone.

OBJECTIVE: Early detection of bone changes following radiotherapy is very important. The aim of this study is to establish a radiographic image analysis method for early detection of radiation-induced trabecular bone changes. METHODS: Thirty-five Wistar rats were used for the experimental model of trabecular bone changes, which were induced by X-ray irradiation with 30 Gy to develop simulated osteomyelitis. Standardized 2.2 x direct magnification radiography was performed immediately prior to X-ray irradiation and once a week for 4 weeks following irradiation. The latent X-ray images were scanned using a computed radiography (CR) system. Mathematical morphological processing was then applied to the CR image data, which allowed the bone trabecular pattern features to be extracted as skeletal binary images. The sensitivity and specificity for detecting early trabecular bone changes in CR images and in skeletal binary images were evaluated. Quantitative analyses with quantum level value (QL value), skeletal pixel percentage (SKP) and star volume analysis (skeletal volume (Vsk), skeletal space volume (Vsp)) were performed. Histopathological examination was also conducted for confirmation of physical changes in the bone. RESULTS: Visual observation of the skeletal binary images provided a higher sensitivity than the CR images. In the quantitative analysis, SKP, Vsk and Vsp values provided higher sensitivity than QL values. CONCLUSION: The results of the present study suggest that a combined radiographic image analysis method using CR, mathematical morphological processing, SKP, Vsk and Vsp can be useful for the early detection of radiation-induced trabecular bone changes.

Morphologically extracted trabecular skeleton superimposed upon digital radiograph structure.

The purpose of this study was to employ a morphological filter to digital X-ray images to extract the morphology of trabecular structures in a clearly understandable visual format. This study compares the trabecular skeleton extracted by a morphological filter to the original digital radiographic image by superimposing the images. A morphological filter (a combination of a single structuring element and a skeleton operation) based on a mathematical morphology theory was used to extract the skeletal pattern of trabecular bone from a digital X-ray image of a human femoral neck. Subset images with different operation numbers (n = 1, n = 2) were obtained, and then each image was superimposed on the original digital radiographic image using the superimpose function of a workstation. The extracted trabecular skeleton pattern was fairly consistent with the trabecular structure seen on the digital image according to the opinion of seven dentomaxillofacial radiologists. In their opinion, the majority of the structural elements were reproduced on the extracted skeleton. However, accurate skeleton elements were not extracted in the region of dense trabecular structure. The morphological filter was able to extract a large portion of the bone trabecular structure as a binary skeletal pattern image from trabecular bone on digital X-ray image, but more work is needed to improve the assessment of dense trabeculae.

Distribution of trigeminal fibers in the primary facial gustatory center of channel catfish, Ictalurus punctatus.

Previous studies in several fishes including catfish, have shown that primary trigeminal nerve (NV) axons terminate not only in the principal and spinal trigeminal nuclei, but in the facial (gustatory) lobes. The present study was undertaken to determine the extent and distribution of trigeminal terminations within the facial lobe (FL) and principal trigeminal nucleus (nVpr) in the channel catfish, Ictalurus punctatus. In order to reveal the distribution of trigeminal fibers, the carbocyanine dye, diI, was applied to the central cut stump of the trigeminal root in isolated, paraformaldehyde-fixed brains. After a diffusion period of 10-90 days, the brains were serially sectioned on a vibratome and examined with epifluorescence. The trigeminal motor nucleus (nVm) and principal sensory nucleus lie near the level of entrance of NV. The majority of primary trigeminal fibers, however, sweep caudally after entering into the brain to form the descending root. At the level of the caudal third of the FL, collaterals emitted by the descending root fibers turn medially and dorsally to terminate in the FL. The trigeminal fibers are coarser than the facial nerve (NVII) fibers which terminate within the same structure. The trigeminal fibers terminate throughout the FL except for the lateral-most lobule which contains the representation of taste buds innervated by the recurrent branch of NVII, i.e., those over the trunk and tail of the animal. These results show that in catfish, the trigeminal input to the primary gustatory complex is restricted to those portions of the nucleus receiving chemosensory inputs from the face and barbels, i.e., the trigeminally innervated sensory fields.

High resolution of DL-tryptophan at high flow rates using a bovine serum albumin-multilayered porous hollow-fiber membrane.

We describe a bovine serum albumin (BSA)-multilayer-adsorbed porous hollow-fiber membrane as a stationary phase that enables chiral separations at a high resolution and high rate. Epoxy-group-containing graft chains were uniformly immobilized on the surface of pores throughout a porous hollow-fiber membrane by radiation-induced graft polymerization. Subsequently, a diethylamino group as an anion-exchange moiety was introduced to the graft chains, which caused the chains to expand toward the interior of the pores due to mutual electrostatic repulsion. The expanding polymer chains provided multilayer binding sites for BSA as a chiral selector. BSA with a degree of multilayer binding of 4 specifically recognized L-tryptophan with a separation factor of 6.6 during permeation by a mobile phase (Tris-HC1 buffer) injected with a racemic solution of DL-tryptophan through the BSA-multilayered porous membrane. In addition, the separation factor was constant irrespective of flow rates of the mobile phase because of negligible diffusional mass-transfer resistance of tryptophan to BSA multilayered by the graft chains.

Chiral separation of DL-tryptophan using porous membranes containing multilayered bovine serum albumin crosslinked with glutaraldehyde.

Bovine serum albumin (BSA) as a chiral ligand was captured uniformly throughout a porous hollow-fiber membrane at a level of 160 mg/g by the polymer chains grafted onto the membrane. BSA was bound in three layers with an end-on orientation to diethylamino groups on the graft chains which expanded from the pore surface towards the pore interior due to mutual electrostatic repulsion. Subsequently, crosslinking of BSA with a 0.025% (w/w) of glutaraldehyde in a Tris-HCl buffer (pH 8) for 4 h was effective in stabilizing the amount of BSA immobilized at a level of 150 mg/g. A solution of DL-tryptophan in a Tris-HCl buffer as a mobile phase permeated the crosslinked-BSA multilayered membrane and produced a chromatogram with a separation factor of 12. BSA leakage was not detected in the mobile phases at various pH values and organic modifiers.

Enhancing effects of binary mixtures of acid with salt on the gustatory neural activity in the clawed toad, Xenopus laevis.

Binary mixtures of acid with salt were observed to produce enhancing effects on oral chemoreceptor responses by recording the activity of the whole glossopharyngeal nerve of the clawed toad (Xenopus laevis). The mixtures of HCl with various inorganic salts elicited responses larger than the sum of the responses to individual component chemicals of each mixture. The results indicated that the mixture response was composed of a large HCI response and a negligibly small salt response. The concentration-response curve for HCl shifted toward lower concentrations due to the presence of NaCl without affecting the maximal response, and the Hill constant for the curve was unchanged, suggesting that the binding affinity of protons increases with an increase in the concentration of the mixed NaCl. The enhancing effects of the mixtures of HCl and various monovalent sodium salts having equal ionic strength showed distinct differences among the anion species of the salts. The responses to the mixtures of HCl with typical chloride salts having cations of various valencies depended on the Cl- ion concentration up to 0.1 M, whereas they depended on ionic strength as well, suggesting a possible role of salt cations in the enhancing effects of the mixtures. Thus, the enhancing effects of salts on the acid responses are interpreted mostly in terms of the interaction of the salt anions on the receptor membrane around the receptor sites for acid. The additive role of the cationic activity of the mixed salts, however, is also involved.

Specificity of glossopharyngeal nerve responses to astringent compounds in Xenopus.

Astringent compounds were applied to oral epithelium of the clawed toad, Xenopus laevis, and rapidly rising and highly sensitive responses could be recorded from the whole glossopharyngeal nerve, but not at all from the trigeminal nerve. The response to 10 mM tannic acid decreased progressively with repetitive application. These responses to tannic acid, however, recovered completely by treating with chemicals capable of forming strong hydrogen and hydrophobic bonds. These chemical bondings are generally recognized as a model for polyphenol (tannin)-protein interactions based on physico-chemical measurements in vitro. The high affinities of these chemicals for tannic acid may be effective in releasing both bonds in the interaction of tannic acid with the receptor molecules. Our results provide in vivo evidence for this model.

Somatotopic organization of the facial lobe of the sea catfish Arius felis studied by transganglionic transport of horseradish peroxidase.

To reveal the somatotopical organization of the facial lobe (FL), a primary medullary gustatory nucleus in the sea catfish Arius felis, the central projections of the peripheral rami of the facial nerve innervating taste buds located across the entire body surface and rostral oral regions were traced by means of horseradish peroxidase neurohistochemistry. The maxillary barbel, lateral mandibular barbel, medial mandibular barbel, and trunk-tail branches project to four different longitudinal columns (i.e., lobules) extending rostrocaudally in the FL. The trunk-tail lobule, which is located dorsolateral to the barbel lobules, lies in the anterior two-thirds of the FL. The tail is represented in a more rostral portion of the trunk-tail lobule than the trunk, indicating that the rostrocaudal trunk axis is represented in the trunk-tail lobule in a posteroanterior axis. The pectoral fin branch ends in an intermediate region of the FL, whereas the hyomandibular, ophthalmic, lower lip, upper lip, and palatine branches terminate in discrete regions of the caudal one-third of the FL. These results reveal a sharply defined somatotopical organization of the FL of Arius and support the hypothesis that the number and lengths of the barbel lobules within the FL of catfishes are directly related to the number and relative lengths of the barbels. An additional subcolumn, the intermediate nucleus of the FL (NIF), which develops in the medioventral region of the caudal two-thirds of the FL, receives projections in a diffuse somatotopical fashion from the barbels, lower lip, and palatine branches. Trigeminal fibers of the barbel and lower lip branches project in a somatotopic fashion to the FL. The present findings suggest that the FL of Arius is highly organized somatotopically to detect, by tropotaxis, precise spatial information concerning taste and tactile stimuli in the environment.

Topographical organization of taste and tactile neurons in the facial lobe of the sea catfish, Plotosus lineatus.

An extraordinary development of the paired medullary facial taste nuclei, the facial lobes, occurs in the sea catfish, Plotosus lineatus. Each of the facial lobes is divided by fiber fascicles into 5 highly distinct lobules or subnuclei, constituting 5 longitudinal columns through the lobe. Extracellular, electrophysiological recordings of neurons within the respective subnuclei of the facial lobe indicate superimposable taste and tactile neural maps organized in a somatotopic manner.

Morphological evidence for a direct projection of trigeminal nerve fibers to the primary gustatory center in the sea catfish Plotosus anguillaris.

The central projections of the ramus mandibularis were examined in the Japanese sea catfish, Plotosus anguillaris by using the technique of transganglionic tracing with horseradish peroxidase (HRP). This ramus receives fibers from both the trigeminal and facial nerves and supplies primarily the two mandibular barbels. Two pathways for a direct trigeminal projection to the facial lobe (FL) were found: one from the main descending root of the Vth nerve (MRDV) to the medial portion of the FL, approximately midway between the rostro-caudal axis of the FL and a second, from deep RDV to the intermediate nucleus (NIF), beneath the medial lobule of the FL. The facial fibers project exclusively onto the medial portion of the FL and the NIF. The results show that fibers of these two cranial sensory nerves supplying the mandibular barbels converge centrally on the medial portion of the FL, indicating that the FL of the Japanese sea catfish is a highly differentiated center for both gustation and somatosensation.

Mechanical sensitivity of the facial nerve fibers innervating the anterior palate of the puffer, Fugu pardalis, and their central projection to the primary taste center.

Mechanical and chemical sensitivity of the palatine nerve, ramus palatinus facialis, innervating the anterior palate of the puffer, Fugu pardalis, and their central projection to the primary taste center were investigated. Application of horseradish peroxidase (HRP) to the central cut end of the palatine nerve resulted in retrogradely labeled neurons in the geniculate ganglion but no such neurons in the trigeminal ganglion, suggesting that the palatine nerve is represented only by the facial component. Tracing of the facial sensory root in serial histological sections of the brain stem suggested that the facial sensory nerve fibers project only to the visceral sensory column of the medulla. Peripheral recordings from the palatine nerve bundle showed that both mechanical and chemical stimuli caused marked responses. Mechanosensitive fibers were rather uniformly distributed in the nerve bundle. Intra-cranial recordings from the trigeminal and facial nerves at their respective roots revealed that tactile information produced in the anterior palate was carried by the facial nerve fibers. Elimination of the sea water current over the receptive field also caused a marked response in the palatine nerve bundle or facial nerve root while this did not cause any detectable responses in the trigeminal nerve root. Single fiber analyses of the mechanical responsiveness of the palatine nerve were performed by recording unit responses of 106 single fibers to mechanical stimuli (water flow), HCl (0.005 M), uridine-5'-monophosphate (UMP, 0.001 M), proline (0.01 M), CaCl2 (0.5 M), and NaSCN (0.5 M). All these fibers responded well to one of the above stimuli; however, most taste fibers did not respond well to the inorganic salts. The palatine fibers (n = 36), identified as mechanosensitive, never responded to any of the chemical stimuli, whereas chemosensitive fibers (n = 70) did not respond to mechanical stimuli at all. The chemosensitive units showed a high specificity to the above stimuli: they tended to respond selectively to hydrochloric acid, UMP, or proline. The responses of the mechanosensitive units consisted of phasic and tonic impulse trains and the sensitivity of the units varied considerably. The results reveal that the facial nerve fibers innervating the anterior palate of the puffer contain two kinds of afferent fibers, chemosensory and mechanosensory respectively, and suggest that the convergence of the tactile and gustatory information first occurs in the neurons of the primary gustatory center in the medulla.

Peripheral and central distribution of major branches of the facial taste nerve in the carp.

The major pathways of the peripheral facial taste system in the carp, Cyprinus carpio, are the maxillary (Max), mandibular (Mand), palatine (Pal) and recurrent nerve rami. The peripheral distribution of the sensory fibers of these branches (B) was determined by means of electrophysiological techniques. Max.B., Mand.B. and Pal.B., each of which arises from the gasserian-geniculate ganglionic complexes, were found to innervate respectively, the upper lip and the adjacent skin, the internal and external surface of the lower lip region, and the upper lip and the anterior palate, ipsilaterally. The recurrent nerve sends fibers mainly via dorsal and ventral branches of the posterior lateral line nerve (NPLL), and a pectoral branch of the occipito-spinal nerve. The dorsal and ventral branches of NPLL innervate respectively, the dorsal fin and the adjacent body surface, and the remainder of the body surface. The pectoral branch supplies the pectoral fin. The central connections of the above branches were also examined by using the techniques of transganglionic tracing with horseradish peroxidase (HRP). HRP was applied to each of the branches, and its penetration of the brainstem was carefully followed. Labeled fibers were observed only in the ipsilateral region of the brainstem. When Max.B or Mand.B. was treated with HRP, labeled fibers were observed in the facial sensory root and in the descending trigeminal root. When Pal.B. was treated, however, they were traced only to the facial sensory root; thus indicating that the former two branches are trigeminofacial complexes and the latter is a pure facial nerve. Labeled fibers for NPLL were found in the facial sensory root as well as in bundles projecting to the lateral line areas. The facial fibers of Max.B. and Mand.B. innervate respectively in the dorsal-intermediate portion of the rostral half of the facial lobe, and in the ventral portion of the caudal half of the lobe. Those of Pal.B. however, cover a large area of the lobe anteroposteriorly except for the dorsal and ventral portions. The recurrent fibers of NPLL and the pectoral B. end in the dorsal-medial portion of the caudal half of the lobe. Thus the results of this study show that there is a topographical relation between the receptive field of the 6 peripheral nerve branches and their locus of representation in the facial lobe. Similarly, that the gustatory system through Pal.B. is represented on the facial lobe in a disproportionately large area compared to that of the other 5 branches.

Distribution of taste buds on the lips and inside the mouth in a minnow, Pseudorasbora parva.

The distribution and abundance of taste buds were quantitatively examined by observing silver impregnated serial sections. The taste buds were widely dispersed on the skin, the lips, the mucosa in the oro-pharyngeal cavity, the esophagus, and the branchial apparatus. The great majority of them was found on the lips and inside the mouth. The external buds were concentrated especially on the outer lips and the adjacent skin, while their number diminished in a caudal direction. Very few were found on the scaled skin. The total number of external buds in a specimen of 6 cm in length was 1,486. The number of taste buds inside the mouth was 6,600. On the inner lips and the palatal organ densities were found to reach over 140 per mm2. High concentrations of taste buds were also found on the gill arches and rakers. These taste buds varied to some extent in size and shape, depending on the thickness of the epithelial layer. It is suggested that the minnow may use the lips, gills and palatal organ as its main taste organs.