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N Okado

Publications and source records attributed to N Okado.

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Immunohistochemical study on development of serotonin-, substance P-, and enkephalin-positive fibers in the rat spinal motor nucleus.

Developmental changes in the coexistence of serotonin and substance P/enkephalin within single fibers of lamina IX of the rat lumbar spinal cord were examined by the use of a double-labelling immunohistochemical technique. On postnatal day (P) 0, 65.0% of immunoreactive varicosities contained only serotonin, and 21.3% of them had both serotonin and substance P. The coexistent ratio of serotonin and substance P in single fibers increased with development: 61.9% of serotonin positive varicosities co-contained substance P on P28, similar to the ratio found in adult animals (67.4%). The ratios of varicosities containing only substance P remained the same from P0 to adult stage (about 15%). Enkephalin positive immunoreactivity was not co-localized with serotonin positive varicosities at any stage of development. Although numerous serotonin positive fibers were found in lamina IX, only a few substance P and enkephalin positive fibers were observed in the same area on P0. The density of serotonin positive varicosities increased slightly by P28, whereas substance P and enkephalin positive fibers increased considerably by this age. Between P28 and the adult stage, the density of serotonin positive fibers decreased by about 50%. The cross sectional area of axonal varicosities containing serotonin- and substance P-like immunoreactivity was similar in both P0 and adult animals, whereas that of enkephalin positive fibers was different. We also examined the coexistence of serotonin and substance P within single neurons of the caudal raphe nuclei in P7 and adult animals, and found that the coexistent ratio significantly increased with development.

Aging

Morphology of sympathetic preganglionic neurons innervating the superior cervical ganglion in the chicken: an immunohistochemical study using retrograde labeling of cholera toxin subunit B.

Preganglionic sympathetic neurons (SPNs) in the chicken were demonstrated immunohistochemically using cholera toxin subunit B (CTb) as a retrograde tracer. After injection of CTb-solution into the superior cervical ganglion, labeled SPNs were mainly found in the ipsilateral sympathetic preganglionic column of Terni (the column of Terni), with only a few in the intermediate zone. They were observed from the caudal half of the 15th cervical segment to the rostral tip of the 3rd thoracic segment. Cell somata of SPNs were loosely packed within the column of Terni, where they had an elliptic shape with the long axis oriented rostrocaudally. In the horizontal plane three kinds of dendrites could be discriminated on the basis of their orientation. Longitudinally oriented dendrites emanated from the rostral and the caudal poles of the SPNs. Medially oriented dendrites were observed to cross the midline and enter the contralateral column of Terni, where they further branched to form a loose dendritic plexus; some extended beyond the lateral limit of the contralateral column of Terni to reach the intermediate zone. Laterally oriented dendrites formed periodically arranged dendritic bundles projecting into the intermediate zone. The present findings provide a detailed account of the dendritic organization of SPNs in the chicken, and suggest that avian SPNs share certain structural features in common with mammalian SPNs.

Animals

Development of serotoninergic system in the brain and spinal cord of the chick.

(1) Development of serotonin positive cells and fibers was immunohistochemically studied by the use of an antibody against serotonin. (2) Serotoninergic neurons were first observed in the immature rohmbencephalon raphe nuclei on embryonic day (E)4, where two clusters of serotonin positive neurons were located: one observed at the rostral part of the rohmbencephalon corresponding to the dorsal raphe nuclei had many serotonin positive cells: the other located at the caudal part of the rohmbencephalon corresponding to the medullary raphe nuclei of the adult animals had only a small number of serotoninergic cells. (3) By E8 the number of serotonin positive cells in the brain stem increased, and virtually all the raphe nuclei found in an adult animal were located. (4) Serotonin positive fibers in the marginal layer reached up to the diencephalon and telencephalon on E6 and E8, respectively. (5) Serotonin positive cells were found beside the midline regions in the ventral part of the spinal cord of the embryonic as well as posthatching chick. (6) Because almost all the serotoninergic fibers in the spinal cord originated from the brain stem raphe nuclei, propriospinal serotonin positive cells were considered as phylogenetic vestiges. (7) Serotoninergic fibers were first found in the marginal layer of the cervical and lumbar spinal cord on E6 and E8, respectively. (8) There was a waiting period of a few days before they penetrated into the mantle layer. (9) Terminal arbolization of the serotoninergic fibers started from late embryonic periods (E16 less than), and was maximized within one week of hatching. (10) Thereafter the density of serotonin positive fibers decreased in all the regions of the spinal cord. (11) Developmental changes of the density of serotonin determined with a high performance liquid chromatography were the same as those determined through immunohistochemistry. Namely the density of serotonin increased linearly from E6 to hatching period, and reached the maximum value one week posthatching. (12( The density of the serotonin in the adult spinal cord was about half of the maximum value. (13) It is to say that the densities of serotonin and serotoninergic fibers transiently increased around one week posthatching. (14) Following the transient increase serotoninergic fibers were eliminated from the neuropil, the fibers were localized in the specific regions of the motor nucleus: motor neuron pools of extensor muscles of the hip joint in the lumbosacral spinal cord.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Identification of Pseudomonas aeruginosa by using a disk of phenanthroline and 9-chloro-9-[4-(diethylamino)phenyl]-9,10-dihydro-10-phenylacridine hydrochloride and by cell agglutination testing with monoclonal antibodies.

To establish a simple identification procedure for Pseudomonas aeruginosa, we developed a disk consisting of phenanthroline and 9-chloro-9-[4-(diethylamino)phenyl]-9,10-dihydro-10- phenylacridine hydrochloride (PC disk). Nine of 248 P. aeruginosa isolates and all other oxidase-positive gram-negative isolates (143 isolates) had clear inhibition zones around the PC disk. Of these nine P. aeruginosa isolates, seven produced a blue, blue-green, or green pigment on Mueller-Hinton agar after 24 h of incubation. The sensitivity and specificity of the PC disk susceptibility test in combination with pyocyanin production were 99 and 100%, respectively. Ten P. aeruginosa isolates showed no agglutination reactions with monoclonal antibodies against P. aeruginosa (96% sensitivity and 100% specificity).

Acridines

Species differences in the distribution and coexistence ratio of serotonin and substance P in the monkey, cat, rat and chick spinal cord.

Serotoninergic raphe-spinal motor neuron projections exhibit wide species differences in both innervation pattern and coexistence of serotonin and substance P. The coexistence ratios vary widely ranging from more than 80% (rat) to less than 1% (chick). Serotonin and substance P positive fibers are also unevenly distributed in the ventral horn of different species: dense clusters of serotonin and substance P positive fibers were preferentially located in the motor neuron pools of extensor muscles of the hip joint (chick) as well as antigravity muscles of the forelimb (cat and rat).

Animals

Serotonin-positive fibers within the spinal motor nucleus of the newborn rat, with special reference to co-localization of substance P.

Developmental changes of serotonin-positive fibers with special reference to co-localization with substance P was examined immunohistochemically in the ventral horn of rat lumbar spinal cord. Only about 20% of serotonin-positive fibers co-labelled with substance P on postnatal day (P) 0. The ratio of co-localization gradually increased, and reached the adult value by P28 (60-70%). Enkephalin-positive fibers were not co-localized with serotonin at any age examined. Although the densities of serotonin, substance P and enkephalin per unit area of the ventral horn gradually increased with development up to P28, the density of serotonin in the adult was decreased compared to P28 animals.

Afferent Pathways

Descending input from the hypothalamic paraventricular nucleus to sympathetic preganglionic neurons in the rat.

The descending projection of the hypothalamic paraventricular nucleus (PVN) to the sympathetic preganglionic neurons (SPNs) in the upper thoracic cord of the rat was studied. PVN-fibers were labeled by anterograde transport of Phaseolus vulgaris leucoagglutinin (PHA-L), while SPNs were retrogradely labeled with cholera toxin subunit B (CTb) which was injected into the superior cervical ganglion. SPNs labeled with CTb were mainly observed in the nucleus intermediolateralis (IML) pars principalis and pars funicularis, and a small number of them were in the nucleus intercalatus (IC) and central autonomic nucleus (CA). SPNs found in the IML had dendrites that projected in various directions. Five types of dendritic projections were noted: medial, rostral, caudal, lateral (including dorsolateral) and ventral. Longitudinal dendritic bundles interconnected each cell cluster in the IML. Medial dendrites of the IML, together with dendrites of the IC and CA, formed transverse dendritic bundles extending from the IML to the central canal. The transverse dendritic bundles disentangled near the midline and formed a loose dendritic plexus in the region just dorsal to the central canal. PVN-fibers labeled with PHA-L were observed primarily in lamina I and intermediate gray (lamina VII). Although varicose PVN-fibers and SPNs coexisted in the IML, the tight packing of the dendritic bundles prevented any clear demonstration of direct contacts between them. On the other hand, PVN-fibers were occasionally found to appose and wind around the primary or secondary dendrites of some SPNs of the CA and IC. These dendrites were studied with varicosities of PVN-fibers for a short length, and terminal boutons of PVN-fibers were also seen to make contact directly with the dendrities. The results of this study substantiated a direct connection between the PVN and SPNs, using a combination of immunohistochemical techniques for PHA-L and CTb. The possible involvement of a direct pathway from the PVN to SPNs in cardiovascular regulation is discussed.

Animals

Coincidence of "ladder-like patterns" in distributions of monoaminergic terminals and sympathetic preganglionic neurons in the rat spinal cord.

A ladder-like pattern of distribution of sympathetic preganglionic neurons (SPNs) was compared with that of monoaminergic terminals in the upper thoracic spinal cord of the rat. SPNs were identified by a retrograde labeling with cholera toxin subunit B (CTb) injected into the superior cervical ganglia of both sides. Monoaminergic terminals were stained immunohistochemically by using antisera raised against 5-hydroxy-tryptamine (5-HT) or dopamine-beta-hydroxylase (DBH). SPNs showing full dendritic arbors were found in all of the sympathetic preganglionic nuclei. They formed a ladder in the horizontal plane. The nucleus intermediolateralis was connected with the central autonomic nucleus by many transverse dendritic bundles. Photomontages of serial sections of material stained alternatively with antisera against CTb and 5-HT or DBH clearly showed a close correlation between SPNs and monoaminergic terminals. There is no transverse dendritic bundle of SPNs without the accompaniment of monoaminergic terminals.

Animals

Immunohistochemical study of tyrosine-hydroxylase-positive cells and fibers in the chicken spinal cord.

Tyrosine hydroxylase (TH)-positive cells and fibers were examined by immunohistochemistry in the chick spinal cord. TH-positive cells, which were located in laminae I, V and X, were most frequently found in the rostral part of the cervical spinal cord, with fewer cells being found in more caudal levels of the spinal cord. TH-positive cells located in lamina X, which were bipolar in shape, were mainly found in regions lateral as well as just ventral to the central canal. They had processes reaching to the central canal. The terminals of these cerebrospinal-fluid-contacting cells were oval in shape, and were most frequently found at the ventral wall of the central canal. There were dense clusters of TH-positive fibers in lamina X. A meshwork-like structure of TH-positive fibers was found over the lateral wall of the central canal. A high density of TH-positive fibers was also found in the medial part of laminae V-VII. In lamina IX, small numbers of TH-positive fibers were observed in the lateral motor column of the brachial spinal cord, and in the medial and lateral motor columns of the lumbosacral spinal cord. However, within the medial motor column of the brachial spinal cord TH-positive fibers were densely distributed around somal as well as dendritic profiles. Similar to our previous observations on serotoninergic fibers. TH-positive fibers were also differentially distributed in the ventral horn of the chicken spinal cord: a high density of TH-positive fibers was localized to specific regions of the spinal motor nucleus.

Animals

Retrograde neuronal labelling by E. coli enterotoxin subunit B.

The present communication reports that subunit B of Escherichia coli heat-labile enterotoxin (LTB) can be utilized as a powerful tracer in neuroanatomical studies. LTB was injected into the limb muscle of the chick, or into the superior cervical ganglion of the rat. Sections were processed with an immunohistochemical technique using an antibody against LTB. After the LTB injection into the muscle, retrogradely labelled motoneurons were found: the entire extent of dendritic arborizations appeared labeled. After the LTB injection into the ganglion, virtually all of the preganglionic neurons were retrogradely labelled in the rat spinal cord.

Afferent Pathways

Distribution patterns of dendrites in motor neuron pools of lumbosacral spinal cord of the chicken.

The morphology of dendritic trees (dendroarchitecture) of motor neurons innervating specific hindlimb muscles (motoneuron pools, MNP) was studied in the chick spinal cord. Motoneurons were labelled by intramuscular injections of horseradish peroxidase conjugated with cholera toxin subunit B. MNPs of posterior iliotibial and femorotibial muscles were located at the dorsolateral part of lateral motor column of lumbosacral segments (LS) 1-4 and 1-3, respectively. Although the dendritic profiles of femorotibialis motoneurons were fewer than those of posterior iliotibialis, these two MNPs had a similar distribution pattern of dendrites. Dendritic profiles were about equally distributed in the gray and white matter. Dendrites from the MNP of posterior iliotibialis radiated in all directions. A large number of dendrites penetrated into the white matter, and some even reached to the subpial regions of the lateral funiculus. One array of dendrites that projected dorsomedialwards extended to the base of the posterior horn. MNPs of both the iliofibularis (LS 4-7) and caudilioflexorius (LS 6-8) had dendritic trees with similar distribution patterns. There were two main arrays of dendritic extensions; one along the dorsal, and another along the ventral border of the lateral motor column. Dendrites from the iliofibularis and caudilioflexorius motoneurons were located more frequently in the white matter than in the gray matter. A large number of dendrites extended in all directions from the MNP of the adductor muscle, which was located in the medial region of lateral motor column of LS 1-2. The distribution of dendrites from a few other MNPs was also examined.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Evidence for two kinds of serotoninergic fibers in the ventral horn of spinal cord of the newly hatched chick.

We observed two types of serotonin-positive (5-HT) fibers in the ventral horn of the lumbar spinal cord of the newly hatched chicken: the first was composed of fine 5-HT fibers, which were increased transiently in the neuropil at 1 week after hatching (transient type); the second type consisted of thick fibers, which were densely localized around motoneuron somata in the motoneuron pools innervating extensor muscles of the hip joint (adult type). Pharmacological perturbation experiments demonstrated that these two types of 5-HT fibers may have different functions: the adult type of fibers may act to maintain an erect standing posture, whereas the transient type may act to induce or facilitate dendrogenesis (or dendritic elongation) of motoneurons. Thus, we concluded that 5-HT fibers may modulate neuronal transmission and serve as a kind of inductive agent for dendritic development in the chick spinal cord.

5,6-Dihydroxytryptamine

Synaptogenesis in the cervical cord of the human embryo: sequence of synapse formation in a spinal reflex pathway.

Synaptogenesis in the cervical cord was studied by light and electron microscopy in human embryos ranging from four to seven weeks of ovulation age. The stage of embryonic development was estimated on the basis of external morphology of embryos and histology of the eye ball with reference to Streeter's horizon. No synapses were found in the cervical cord of the embryo at Streeter's horizon XIV (8 mm; estimated ovulation age, 28-30 days). A small number of axodendritic synapses appear in the motor neuropil of the cervical cord at Streeter's horizon XVII (14 mm; estimated ovulation age, 34-36 days). Since no primary afferents are demonstrated to reach the motor neuropil at this stage (the premotile period), these synapses are considered to be formed between interneurons and motor neurons. On the other hand, the formation of synapses outside the motor neuropil of the cervical cord was recognized at Streeter's horizon XX (22 mm; estimated ovulation age, 40-42 days), which corresponded to the period of onset of the precocious reflex, but not by horizon XIX (18 mm; estimated ovulation age, 38-40 days). The first axosomatic synapses were found in the motor neuropil at Streeter's horizon XVII (estimated ovulation age, 34-36 days). The present study suggests that the formation of synapses between interneurons and dendrites of spinal motor neurons precedes that of synapses between interneurons and collaterals of primary afferents. This sequence of synaptogenesis is in agreement with that reported in earlier studies with silver stain methods.

Gestational Age