Biomedical subjects
Y Shigenaga
Publications and source records attributed to Y Shigenaga.
Morphology and synaptic connections of slowly adapting periodontal afferent terminals in the trigeminal subnuclei principalis and oralis of the cat.
Previous studies suggest that sensory information from primary afferent fibers is processed in a distinct manner in the individual subnuclei of trigeminal sensory nuclear complex. The present study has addressed this issue by using intra-axonal labeling with horseradish peroxidase to examine the ultrastructure and synaptic organization of axon terminals from slowly adapting (SA) periodontal afferents in the ventral subdivision (Vpv) of principalis and the rostro-dorsomedial part (Vo.r) of oralis. Our observations are based on complete or near-complete reconstructions of 139 synaptic boutons in Vpv and 105 in Vo.r. All the labeled boutons contained clear, spherical, synaptic vesicles and were presynaptic to unlabeled dendrites, and they were frequently postsynaptic to unlabeled axon terminals containing pleomorphic synaptic vesicles (P-endings). The P-endings frequently formed axodendritic synapses on dendrites which received axodendritic synapses from labeled boutons (synaptic triads). On the basis of the number of contacts, synaptic arrangements associated with the labeled boutons could be subgrouped into simple (one or two contacts), intermediate (three or four contacts), and complex (five or more contacts) types. The labeled boutons varied from round to elongated forms with smooth to more irregular or scalloped contours. The boutons with scalloped contour were much more frequent in the complex type. The boutons of the intermediate type were significantly smaller than the complex type and larger than the simple type. The SA periodontal afferent terminals participated in each type of synaptic arrangements in Vpv, but were mostly of the simple type in Vo.r. The size of labeled boutons was significantly larger in Vpv than in Vo.r. The total number of axodendritic and axoaxonic contacts per labeled bouton was significantly higher in Vpv than in Vo.r. Another difference was the more frequent occurrence of synaptic triads in Vpv than in Vo.r. These observations provide evidence that sensory information from primary afferent fibers is processed in a different manner in the two subnuclei.
Two major types of premotoneurons in the feline trigeminal nucleus oralis as demonstrated by intracellular staining with horseradish peroxidase.
Previous studies suggest that neurons in the dorsomedial subdivisions of trigeminal nucleus oralis (Vo) may contribute to reflex control of jaw movements and to modulation of sensory information. The present study has addressed this possibility by the use of intracellular staining with horseradish peroxidase of physiologically identified neurons in Vo to examine functional and morphological properties of these neurons. Of 14 labeled neurons, eight had axon collaterals terminating exclusively in the dorsolateral subdivision of the trigeminal motor nucleus (DL neurons) and four in its ventromedial subdivision (VM neurons); axon collaterals of two neurons were not traced. Both groups of neurons sent terminal arbors into other nuclei of the lower brainstem. The DL neurons were distinguishable from the VM neurons in their receptive field (RF) location, neuronal position, somadendritic architecture, and projections to other brainstem nuclei. All neurons, except for two that were exclusively activated by noxious stimuli applied to the tongue, were responsive to light mechanical stimulation of peri- and intraoral structures. The RFs of the DL neurons were located in more posterior oral structures than those of the VM neurons. The RF of nearly all low-threshold DL neurons was located in the maxillary region, and that of the VM neurons, in contrast, involved the mandibular region. The VM neurons were located medial or ventral to the DL neurons. The soma size of the VM neurons was significantly larger than that of the DL neurons. Dendritic arbors of both groups could be separated into medial and lateral components. The ratio of the dendritic transverse areas in the medial vs. lateral component was significantly higher in the VM neurons than in the DL neurons. The DL neurons also issued collaterals that terminated in larger brainstem areas than those of the VM neurons. These observations provide new evidence on the morphological and functional properties of Vo neurons that contribute to reflex control of jaw and facial movements and modulation of sensory information.
Central terminations of low-threshold mechanoreceptive afferents in the trigeminal nuclei interpolaris and caudalis of the cat.
Previous studies indicate that vibrissa, nonvibrissa, guard hair, hairy skin, and periodontal ligament afferents give rise to morphologically distinct terminal arbors in the trigeminal nuclei principalis (Vp) and oralis (Vo) in the cat. The present study describes the extent to which morphological and functional relationships exist in the nuclei interpolaris (Vi) and caudalis (Vc). Twenty-two fibers were physiologically characterized and stained by intra-axonal HRP injection techniques. The fast adapting (FA) vibrissa (VF) afferents gave rise to compact and circumscribed arbors in Vi and Vc. These tended to be larger in Vc than in Vi. The slowly adapting (SA) vibrissa (VS) afferents in Vi and Vc had more widespread and larger arbors than those of the VF afferents. The VS arbors in Vc tended to be larger and less circular than those in Vi. Guard hair (GH) afferents gave rise to circumscribed arbors in both nuclei, but those in Vc tended to have larger and more circular arbors than those in Vi. Down hair (DH) afferents gave rise to small, circumscribed arbors or a few distinct patches of boutons within a small area in Vi; arbors in Vc were less extensive and "stringy." Unlike other afferents, DH arbors were larger in Vi than in Vc, but smaller than those of GH afferents in either nuclei. The SA hairy skin (SS) afferents had arbors that were widespread with a few distinct patches of boutons; the arbors in Vc were larger than those in Vi. The arbors of SS afferents were smaller than those of VS and GH afferents in both nuclei. Like GH afferents, FA periodontal ligament (PF) afferents gave rise to large and circumscribed arbors in Vi, although the arbors in Vc were smaller and less dense. The present study demonstrated significant functional and morphological relationships for primary afferents in Vi and Vc, thus suggesting that sensory information from each of the distinct fiber or functional classes is processed in a characteristic manner in the individual nuclei.
NADPH-diaphorase in the spinal trigeminal nucleus oralis and rostral solitary tract nucleus of rats.
NADPH-diaphorase histochemical staining demonstrated a distinct neural group that might synthesize nitric oxide in the lower brainstem of rats. The NADPH-diaphorase stain revealed a Golgi-like network in the dorsomedial spinal trigeminal nucleus oralis and rostrolateral solitary tract nucleus, whereas this network was more dense in the latter nucleus. The distribution of NADPH-diaphorase-positive neurons in these areas overlapped with parts of central terminations from the chorda tympani nerve, as demonstrated with transganglionic transport of wheatgerm agglutinin conjugated horseradish peroxidase. The number of NADPH-diaphorase-positive neurons changed after chorda tympani nerve lesion relative to the contralateral side. The control value (%) was 106.0 +/- 4.9 (mean +/- S.E.M.). One hour after the nerve lesion, the value increased to 115.2 +/- 9.1 (P > 0.05). It then decreased to 83.9 +/- 5.2 two days after the lesion (P < 0.05), and remained at this reduced level for one or two weeks, 83.2 +/- 3.0 (P < 0.01) and 83.7 +/- 2.3 (P < 0.01), respectively. This statistically significant reduction recovered to control level 103.4 +/- 2.9 four weeks after the lesion. These results show that NADPH-diaphorase-positive neurons in the lower brainstem could be regulated trans-synaptically by primary afferents, possibly gustatory inputs.
Synaptic connections of a periodontal primary afferent neuron within the subnucleus oralis of the cat.
The central axon of a primary afferent neuron that responded to light mechanical stimulation of the lower premolar teeth in a fast adapting fashion was intra-axonally injected with horseradish peroxidase in the cat. The labeled terminals within the rostrodorsomedial (Vo.r) and dorsomedial (Vo.dm) parts of subnucleus oralis were examined electron microscopically. The labeled ending had pale axoplasm, contained clear spherical synaptic vesicles, and formed multiple synapses with dendrites and/or unlabeled axonal endings with pleomorphic vesicles (P-endings). In these synaptic contacts, the labeled primary ending was presynaptic to dendrites and postsynaptic to P-endings. Labeled endings simultaneously synapsing with both dendrites and P-endings were more frequent in Vo.dm (28%) than in Vo.r (8.3%).
Nucleus ambiguus of the rabbit: cytoarchitectural subdivision and myotopical and neurotopic representations.
The cytoarchitectural subdivisions of the nucleus ambiguus of the rabbit and its myotopical and neurotopical representations were investigated with HRP labeling. The nucleus was subdivided into the compact cell group (CoG), the medial and lateral scattered cell groups (SGm and SGl), and the diffuse cell group (DiG). The CoG was formed by esophageal, pharyngeal constrictor, and palatal motoneurons in the rostral half of the nucleus. The SGm and SGl were located medial and lateral to the CoG, respectively, in the rostral one-third of the nucleus. Stylopharyngeal and cricothyroid motoneurons were located in the most rostral one-fifth of the SGm and the remaining four-fifths, respectively, whereas the SGl was not labeled with HRP injections into the palatal, pharyngeal, esophageal, and laryngeal muscles. The DiG was formed by recurrent laryngeal motoneurons in the caudal two-thirds of the nucleus. Neurons of origin for the glossopharyngeal nerve occupied the stylopharyngeal region, with a few of them scattered in the CoG and SGl. Neurons giving rise to axons in the superior laryngeal nerve occupied the cricothyroid region, with a few of them scattered in the pharyngeal constrictor region; whereas the pharyngeal vagal branch originated from the pharyngeal constrictor and palatal regions. Neurons of the DiG, SGl, and esophageal region contributed to the infranodosal vagus nerve; esophageal fibers of the recurrent laryngeal nerve originated from the dorsal esophageal region. Laryngeal fibers of the recurrent laryngeal nerve originated from the DiG, the caudal neurons of which had axons traversing the cranial accessory root.
Effect of soft diet and aging on rat masseter muscle and its motoneuron.
To determine the effect of a soft diet and aging on the masticatory motor unit, we investigated the morphologic and metabolic properties of the superficial masseter muscle and its motoneurons in rats. Twenty rats were divided into four groups of five rats: rats fed a hard diet until 4 months after birth (hard, young), rats fed a soft diet until 4 months after birth (soft, young), rats fed a hard diet until 22 months after birth (hard, old), and rats fed a soft diet until 22 months after birth (soft, old). The diameter of the fast-twitch oxidative glycolytic muscle fiber was significantly smaller in the soft than the hard, and in the old than the young groups. The glycolytic enzyme (phosphofructokinase) activity of the muscle was significantly weaker in the old than the young group. There was no significant difference in soma diameter of the motoneurons between the soft and hard group, while the diameter was significantly larger in the old than in the young group. There was no significant difference in NADH-diaphorase activity of the motoneurons between the soft and hard group, while significantly less activity was demonstrated in the old than in the young group. The reduction in motor unit activity caused by the soft diet is considered to influence the morphologic and metabolic properties in the superficial masseter muscle but not in its motoneurons. The reduction in the oxidative enzyme activity of motoneurons with aging may occur regardless of the reduction in motor unit activity.
The central projections of the monkey tooth pulp afferent neurons.
Transganglionic transport of horseradish peroxidase conjugated to wheatgerm agglutinin (HRP:WGA) entrapped in hypoallergenic polyacrylamide gel was used to study the patterns of termination of primary afferents that innervate the upper and lower tooth pulps within the trigeminal sensory nuclear complex (TSNC) of the monkey. HRP:WGA injections were also made into the lower incisors and molars, in order to examine the topographic arrangement of pulpal afferent projections. HRP-labeled pulpal afferents innervating lower and upper teeth projected ipsilaterally to the rostral subnucleus dorsalis (Vpd) and caudal subnucleus ventralis (Vpv) of the nucleus principalis (Vp); the rostrodorsomedial (Vo.r) and dorsomedial (Vo.dm) subdivisions of the nucleus oralis (Vo); the dorsomedial subdivision of the nucleus interpolaris (Vi); and laminae I-II and/or V of the nucleus caudalis (Vc) at its rostralmost level. The HRP-labeled terminals from upper and lower pulpal afferents formed a rostrocaudal column from the midlevel of Vp to the rostral tip of Vc. The label in Vp and Vo was considerably dense, but the column of terminals was interrupted at the Vpd-Vpv transition. The label in Vi and Vc was much less dense compared to that in the rostral nuclei, and the column of terminals was interrupted frequently. The representation of the upper and lower teeth in TSNC was organized in a somatotopic fashion that varied from one subdivision to the next, though their terminal zones overlapped within Vpd. The upper and lower teeth were represented in Vpv, Vo.r, Vo.dm, Vi, and Vc in a ventrodorsal, dorsoventral, lateromedial, lateromedial, and lateromedial sequence, respectively. Topographic arrangement was also noticed for the projections of pulpal afferents from the lower incisors and molars: The representations of the lower incisors and molars in Vpv, Vo.r, Vo.dm, Vi, and Vc were organized in a lateromedial, dorsoventral, ventrodorsal, ventrodorsal, and lateromedial sequence, respectively. The present results indicating sparse projections from pulpal afferents in the monkey's Vc are in good correspondence with a clinical report that trigeminal tractotomy just rostral to the obex has no significant effect on dental pain perception in patients. Furthermore, the present study indicates that projection patterns of pulpal afferents--which include the termination sites, the density of terminations between nuclei, and topographic arrangement--differ among animal species.
An HRP study of the location of the motoneurons supplying the tensor veli palatini muscle of the rabbit.
Location of the motoneurons supplying the tensor veli palatini muscle of the rabbit was examined with the retrograde labeling technique following intramuscular injection of HRP. Labeled motoneurons were ipsilaterally located in the ventral or ventromedial portion of the rostral two-thirds of the motor trigeminal nucleus at the level of about 6.0 to 8.5 mm rostral to the obex. The location of the labeled motoneurons was ventromedial to the region supplying the masseter, the temporalis, and the medial pterygoid muscles and ventral to the region supplying the anterior digastric and the mylohyoid muscles, the location which coincided with the lateral pterygoid region. The labeled motoneurons were scattered around or in this region.
Synaptic connections of a low-threshold mechanoreceptive primary neuron within the trigeminal subnucleus oralis.
The central axon of a primary afferent neuron that responded to indentation of the glabrous skin of the lower lip in a slowly adapting fashion was intra-axonally injected with horseradish peroxidase. The labeled terminal within the subnucleus oralis was examined electron microscopically. The labeled ending had a pale axoplasm and contained clear spherical synaptic vesicles. The labeled ending formed a synaptic triad with a dendrite and an unlabeled axonal ending with pleomorphic vesicles (a mixture of oval, flattened and dense core vesicles). The labeled primary ending was presynaptic only to the dendrite, while the unlabeled ending was presynaptic to both the dendrite and the labeled primary ending.
A morphological study on glutamate-induced swelling of cultured astrocytes: involvement of calcium and chloride ion mechanisms.
Morphological changes in cultured astrocytes exposed to L-glutamate (Glu) were examined light and electron microscopically. The treatment with 0.1 mM Glu for 60 min caused marked swelling of the cells, which was characterized by reduction in staining of cytoplasm with Toluidine blue, disappearance of the cytoplasmic granular ground substances, swollen mitochondrion and nucleus, and dispersed chromatin. The above changes were prevented by the removal of Na+, Ca2+ or Cl- from the incubation medium for Glu treatment. However, the Glu treatment in a Cl(-)-free medium caused conspicuous aggregation of 10 nm filaments.
Location of the motoneurons supplying the rabbit pharyngeal constrictor muscles and the peripheral course of their axons: a study using the retrograde HRP or fluorescent labeling technique.
The location of the motoneurons supplying the rabbit pharyngeal constrictor muscles (the superior and the middle constrictors, and the thyropharyngeus and the cricopharyngeus; the last two collectively compose the inferior constrictor) was investigated with intramuscular injection of HRP or the fluorescent tracer nuclear yellow into the individual muscles. Moreover, the peripheral course of their axons was investigated by injection of HRP into all of the pharyngeal constrictors in conjunction with intracranial severing of either the vagus or the glossopharyngeal nerves. The pharyngeal constrictor motoneurons were ipsilaterally located within a subdivision of the nucleus ambiguus which is formed by a compact arrangement of the smallest neurons of the nucleus and situated in the rostral half of the nucleus. We named that subdivision the compact cell group (CoG). Axons of the pharyngeal constrictor motoneurons traversed the vagal rootlets. The rostrocaudal extent of the pharyngeal constrictor motoneurons covered almost the entire length of CoG at a level from about 500 to 2,900 microns rostral to the obex, with their number being most numerous in the middle one-third level of the CoG. Although the motoneurons of the superior constrictor, those of the middle constrictor, and those of the thyropharyngeous and the cricopharyngeus overlapped considerably in location, they tended to be arranged rostrocaudally in that order. At the middle one-third level of the CoG, where the CoG is subdivided into dorsomedial and ventrolateral subgroups of neurons, the superior and the middle constrictor motoneurons were confined to the medial portion of the dorsomedial subgroup, while the inferior constrictor motoneurons were distributed throughout its entirety.
Difference in central projection of primary afferents innervating facial and intraoral structures in the rat.
Transganglionic transport of horseradish peroxidase-wheat germ agglutinin conjugate was used to study the central projection of primary afferent neurons innervating facial and intraoral structures. The examined primary neurons innervating the facial structures were those comprising the frontal and zygomaticofacial nerves and those innervating the cornea, while the primary neurons innervating the intraoral structures included those innervating the mandibular incisor and molar tooth pulps and those comprising the palatine nerve. The primary afferents innervating the facial structures project to the lateral or ventral parts of the trigeminal principal, oral and interpolar subnuclei, and to the rostral cervical spinal dorsal horn across laminae I through V, with a greater proportion being directed to the spinal dorsal horn. The primary afferents innervating the intraoral structures terminate in the dorsomedial subdivisions of the trigeminal principal, oral and interpolar subnuclei, and in laminae I, II, and V of the medial medullary dorsal horn, with a much denser projection being distributed to the rostral subnuclei. In addition to the above brain stem trigeminal sensory nuclear complex, they project to the supratrigeminal nucleus, caudal solitary tract nucleus, and paratrigeminal nucleus. These observations agree with previously reported data that the central projection of trigeminal nerve is organized in different manners for the facial and intraoral structures. Furthermore, the present findings in conjunction with our previous studies clarify that the central projection of primary afferents from the facial skin is organized in a clear somatotopic fashion and that the terminal fields of primary afferents from the intraoral structures extensively overlap in the brain stem trigeminal nuclear complex particularly in its rostral subdivisions. The central mechanism of trigeminal nociception is discussed with particular respect to its difference between the facial and intraoral structures.
Morphology of central terminations of low-threshold trigeminal primary afferents from facial skin in the cat--intra-axonal staining with HRP.
Intra-axonal recording and horseradish peroxidase (HRP) injection techniques were employed to examine the response properties of low-threshold mechanoreceptive afferents and the morphological characteristics of their axon arbors in the main sensory nucleus (Vp) and oral nucleus (Vo). Thirteen afferents were characterized and recovered. One gave fast or rapidly adapting (FA) and 3 slowly adapting (SA) responses to mystacial vibrissa deflection, 5 were sensitive to deflection of non-vibrissae hairs or hair (4 were guard hair afferents and the other responded to deflection of a long hair in slowly adapting fashion) and two were responsive to indentation of the hairy skin. The remainder were responsive to indentation of the glabrous skin on the lower lip: one was of FA type and the other of SA type. All of the axons had bifurcating fibers that ascended in the ascending tract (ascending fiber) and descended in the trigeminal spinal tract (descending fiber). The main collaterals given off from the ascending fiber and rostral segment of the descending fiber terminated in the Vp, and the other collaterals from the descending fiber projected to the Vo. Terminal arbors produced by the main collaterals formed a rostrocaudally continuous column, but generally the adjacent arbors did not overlap except when pairs of collaterals arose near each other on the ascending and descending fibers. Projections of collaterals to Vp and Vo were organized topographically. The head was represented in an inverted fashion with its anteroposterior axis in a mediolateral sequence, but the lower glabrous lip was represented more dorsally than the other mandibular facial regions. Vibrissa afferents formed a rostrocaudally continuous, densely packed terminal column throughout the length of Vp and Vo. SA vibrissa afferents gave rise to more dense and roundish arbors in Vp than the FA afferent, while the Vo.c arbors were more compact and smaller than those of the FA afferent. Guard hair afferents had arbors that were highly variable throughout the nuclei and were characterized by less developed arbors in Vp than in Vo. Unlike vibrissa afferents, hairy skin afferents gave rise to sparse and widespread arbors characterized by a string-like appearance, while the Vo collaterals were more stringy. Facial lip afferents were characterized by a great difference in collateral morphology between FA and SA type.(ABSTRACT TRUNCATED AT 400 WORDS)
Two types of jaw-muscle spindle afferents in the cat as demonstrated by intra-axonal staining with HRP.
Intra-axonal records and horseradish peroxidase (HRP) injection techniques were employed to define the response properties of the jaw-closing muscle spindle afferents in the trigeminal mesencephalic nucleus (Vmes) and their morphological characteristics. The axonal trajectories of 9 spindle afferents from the masseter and 4 afferents from the temporalis were recovered for detailed analyses. Of 13 afferents, 6 cell bodies were stained and they were located at the rostrocaudal mid-levels of the Vmes. The central courses of the stem fibers were organized in a similar manner to the Vmes periodontal afferent nerves with the exception that peripheral (P) fibers of all spindle afferents passed through the trigeminal motor tract and root. On the basis of collateral terminal arborizations, the Vmes spindle afferents could be classified into two types: type I (n = 6) and type II (n = 7). Type I afferents sent their collaterals into the trigeminal motor nucleus (Vmo), intertrigeminal region (Vint) and juxtatrigeminal region (Vjux), but collaterals from the two neurons also projected to Vmes and the nucleus oralis (Vo). The collaterals from type II afferents formed their terminal arbors in the supratrigeminal nucleus (Vsup) in addition to the Vmo, Vint and Vjux, but collaterals from one neuron also projected to the Vo. In type I afferents, terminal arbors encompassed the whole Vmo including jaw-closing motoneurons. In contrast, boutons from type II afferents were restricted to a few small portions within the Vmo in proximity to its lateral and dorsal boundaries. The diameters of the united (U), central (C) and peripheral (P), fibers were larger in type I than type II afferents; those of the U fibers were statistically significant. Any differences between the two distinct types were not found in the response pattern to the sustained jaw opening. These results suggest that the difference of primary and secondary muscle-spindle afferent nerves is reflected in a distinctive morphology in the terminal arborizations and in the diameters of united fibers rather than the response patterns in deeply anesthetized cats.
Physiological and morphological characteristics of periodontal mesencephalic trigeminal neurons in the cat--intra-axonal staining with HRP.
Intra-axonal recording and horseradish peroxidase (HRP) injection techniques were employed to define the response properties of periodontal mechanoreceptive afferents originating from the trigeminal mesencephalic nucleus (Vmes) and their morphological characteristics. The periodontal Vmes neurons were classified into two types: slowly adapting (SA) and fast adapting (FA) types. The central terminals of 7 SA and 4 FA afferents were recovered for detailed analyses. The whole profile of SA and FA neurons were unipolar in shape and their cell bodies were located in the dorsomedial parts of the Vmes. The united (U) fiber traveled caudally from the soma to the dorsolateral aspect of the trigeminal motor nucleus (Vmo), where it split into the peripheral (P) and C fibers with a T- or Y-shaped appearance. The P fiber joined the trigeminal sensory or motor tract. The C fiber descended caudally within Probst's tract. All 3 stem fibers issued main collaterals. The main collaterals of all neurons examined formed terminal arbors in the supratrigeminal nucleus (Vsup) and all but two SA neurons projected to the intertrigeminal region (Vint), while the projections to other nuclei of the trigeminal motor nucleus (Vmo), juxtatrigeminal region (Vjux), main sensory nucleus (Vp) and oral nucleus (Vo.r) differed between SA and FA afferents and between neurons of the same type. The SA and FA neurons were classified into three and two subgroups, respectively. The major differences in central projections between the two types were that all the FA neurons projected to the Vp or Vo.r but none of SA type and this relation was reversed in the projection to the Vjux, and that more than half of SA neurons projected to Vmo but only one FA neuron to the Vmo. The Vmes neurons which sent their collaterals into the Vmo had the P fiber passing through the tract of the trigeminal motor nerve. The average size of somata and mean diameters of U fibers and main collaterals from C fiber were significantly larger in SA neurons than FA neurons. The average size of fiber varicosities became smaller in the following nuclei, Vmo, Vsup, Vp, Vint and Vo.r, but not significant between the two functional types. The functional role of the periodontal Vmes afferents to jaw reflexes was discussed particularly with respect to their central projection sites in the brainstem nuclei.