New human papilloma virus isolated from epidermodysplasia verruciformis lesions.

Human papilloma virus (HPV) was isolated from red plaques of a patient (N. F.) with epidermodysplasia verruciformis. Electron microscopic examination showed characteristic particles of papilloma virus as icosahedrons about 45 nm in diameter. DNA was extracted from these particles, and closed-circular DNA (Form I) was purified by centrifugation in CsCl containing ethidium bromide. The molecular weight of the DNA was about 5.0 x 10(6). A physical map of the HPV DNA was constructed using several restriction enzymes. The restriction endonuclease cleavage pattern of the HPV DNA was different from those of other types of HPV reported thus far, suggesting that the isolate was a new, as yet unclassified, HPV.

Developmental appearance of oligodendrocytes in the embryonic chick retina.

The axons of the optic nerve layer are known to be myelinated by oligodendrocytes in the chick retina. The development of the retinal oligodendrocytes has been studied immunohistochemically with antibodies against oligodendrocyte lineage: monoclonal antibodies O4 and O1, and an antibody against myelin basic protein. O4 positive (O4+) cells were first detected in the retina on the tenth day of incubation (embryonic day (E)10, stage 36). The labeled cells were located in the optic nerve layer close to the optic fissure. Most were unipolar in shape, extending a leading process with a growth cone toward the periphery of the retina. By E12, unipolar O4+ cells had spread to the middle of the retina. Many O4+ cells close to the optic fissure showed radial arrangement with extension of processes toward the inner limiting membrane. O1+ oligodendrocytes were first observed in the E14 retina positioned just above (interiorly to) retinal ganglion cells. These labeled cells extended fine processes in the optic nerve layer. Limited numbers of myelin basic protein-positive cells were present by E16 and located interiorly to the retinal ganglion cells. In addition to the oligodendrocyte in the optic nerve layer, a limited number of O4+ cells were observed in the inner nuclear layer by E14, and they became O1+ by E18. Furthermore, explant culture experiments showed E10 to be the youngest stage at which the retina contained oligodendrocyte precursors. An intraventricular inj ection of fluorescent dye 1,1',dioctadecyl-3,3,3',3-tetramethylindocarbocyanine perchlorate (DiI) at E6 yielded O4+/DiI+ cells in the retina at E10, which provided direct evidence to support migration of oligodendrocyte precursor into the retina. The present results demonstrated the sequential appearance of the cells of oligodendrocyte lineage and the detailed morphology of the developing oligodendrocytes in the retina. These morphologic features strongly suggested that retinal oligodendrocytes were derived from the optic nerve and spread by migration through the optic nerve layer.

Topographical organization of projections from the subiculum to the hypothalamus in the rat.

The projections from the subiculum to the hypothalamus were comprehensively examined in the rat by using the anterograde Phaseolus vulgaris leucoagglutinin (PHA-L) and retrograde cholera toxin B subunit (CTb) methods. Tracing of efferents with PHA-L indicated that the medial preoptic region received projection fibers from the temporal two-thirds of the subiculum, whereas the anterior, tuberal, and mammillary regions received those from the full longitudinal extent of the subiculum. The subicular projections to the anterior and tuberal hypothalamic regions were also found to be organized in a topographical manner such that the temporal-to-septal axis of origin in the subiculum determined a ventromedial-to-dorsolateral axis of termination in the medial zone of the hypothalamus: Massive labeled fibers from the temporalmost part of the subiculum terminated in the subparaventricular zone and its caudal continuum around the dorsal and medial aspects of the ventromedial nucleus, and those from progressively more septal parts terminated in progressively more dorsolateral parts of the medial zone. In addition, the temporal-to-septal axis of origin in the subiculum tended to determine a medial-to-lateral axis of termination in the preoptic region as well as a ventral-to-dorsal axis of termination in the mammillary region. Furthermore, the temporal-to-septal axis of origin in the septal two-thirds of the subiculum corresponded to a ventrolateral-to-dorsomedial axis of termination in the medial mammillary nucleus. The topographical projections from the subiculum to the medial zone of the hypothalamus were confirmed by CTb experiments, representatively in the subicular projections to the anterior hypothalamic region. These results suggest that different populations of neurons existing along the longitudinal axis of the subiculum may exert their influences on the execution of different hypothalamic functions.

Characteristics of distribution of peptide-containing nerve fibres in the atrioventricular valves of the rat.

The distribution of vasoactive intestinal polypeptide-, neuropeptide Y-, and calcitonin gene-related peptide-immunoreactive nerve fibres was investigated in the atrioventricular valves of the rat. These nerve fibres were visualized by immunostaining of whole-mount preparations by the avidin-biotin-peroxidase complex method. Vasoactive intestinal polypeptide-immunoreactive nerve fibres were observed mainly in the anterior cusp of the mitral valve and, to a lesser extent, in the medial cusp of the tricuspid valve. Numerous neuropeptide Y-immunoreactive nerve fibres were found covering all of the cusps. Both types of peptidergic nerve fibre formed dense networks that consisted of interlacing and anastomosing nerve fibres. Calcitonin gene-related peptide-immunoreactive nerve fibres were seen in every cusp, but did not form a fine network. These results provide detailed anatomical information for evaluation of the possible roles of each type of peptide-containing nerve fibre in the function of atrioventricular valves.

Nerve fibres containing neuropeptide Y in the atrioventricular valves of Japanese monkey and rat; a light and electron microscopic study.

Dense distribution of varicose fibres containing neuropeptide Y-like immunoreactivity (NPY-LI) was found in the atrioventricular valves of the Japanese monkey, and moderately in the rat. The immunoelectron microscopy using immunogolds resulted in the localization of NPY-LI within the dense-cored vesicles which existed with the small clear vesicles in the unmyelinated axons near the endocardium. These NPY-LI-containing fibres may participate in regulation of vasomotor role or other functions of the atrioventricular valves.

Removal of mesenteric gastrinoma: a case report.

A 37-year-old man with a primary mesenteric gastrinoma is reported. A parathyroid adenoma had been removed 13 years ago. Six years earlier the patient underwent an emergency distal gastrectomy because of massive bleeding from a duodenal ulcer. Two months later stomal ulcers developed associated with hypergastrinemia. No gastrinoma was detected and total gastrectomy was performed. The fasting plasma level of gastrin was stable in the range from 580 to 920 pg/ml for the following 5 years. However, the level was found to abruptly increase to 4125 pg/ml. The level increased progressively to 11383 pg/ml within 1 year. A gastrinoma was identified in the jejunal mesenterium, and it was completely removed. After surgery the plasma level of gastrin decreased below the limit of the assay, and the paradoxical response to secretin was no longer observed.

A nigro-rubro-bulbar pathway to the parvicellular reticular formation in the rat.

A possible pathway from the substantia nigra pars reticulata (SNr) to the parvicellular reticular formation (RFp) via the red nucleus (RN) was examined light and electron microscopically by combining anterograde and retrograde tracing techniques. After contralateral injections of biotinylated dextranamine (BDA) into the dorsolateral part of the SNr and cholera toxin B subunit (CTb) into the RFp, many CTb-labelled neurones were distributed contralaterally in the dorsal part of the RN, where numerous BDA-labelled axon terminals originating from the ipsilateral SNr were found. After contralateral injections of BDA into the dorsolateral part of the SNr and wheat germ agglutinin-horseradish peroxidase (WGA-HRP) into the RFp, ipsilateral axons labelled with BDA were found to make synaptic contacts with the somata and dendrites of contralateral neurones labelled with WGA-HRP in the dorsal part of the RN.

Enhancement of neurite outgrowth by the soluble form of human L1 (neural cell adhesion molecule).

L1, a neural cell adhesion molecule, is involved in neurite outgrowth, migration and fasciculation. Although L1 is a membrane glycoprotein expressed on neural cells, the soluble form of L1 is generated in vivo by proteolysis. In the present study, a stable transfectant of Chinese hamster ovary (CHO) cells secreting human L1 without cytoplasmic and membrane spanning domains was generated, and the function of the secreted L1 was examined. Explants from embryonic chick brain stem were cultured on a substrate coated with polyethylenimine (PEI) alone, on substrate-bound L1 or in medium containing soluble L1. The neurites induced by L1, both the substrate-bound form and the soluble form, were 2-3 times longer than those cultured on PEI. The ability of the soluble L1 to induce neurite formation was slightly greater than that of the substrate L1. The present results demonstrated that neurite outgrowth was induced not only by substrate-bound L1 but also by soluble L1. Soluble L1 could be a pharmaceutical candidate for the promotion of nerve regeneration.

Projections from the red nucleus to the parvicellular reticular formation and the cervical spinal cord in the rat, with special reference to innervation by branching axons.

After biotinylated dextranamine injection into the dorsal part of the red nucleus (RN) in the rat, labeled axons were distributed contralaterally in the lateral tegmental field including the parvicellular reticular formation (RFp), and ipsilaterally in the medial reticular formation. In the cervical spinal cord, labeled axons were present bilaterally with a contralateral dominance mainly in laminae V-VI and the dorsal part of laminae VII. After ipsilateral injections of rhodamine dextranamine, Fluoro-ruby (FR) into the RFp and Fluoro-gold (FG) into the upper cervical spinal cord, a population of FR-labeled neurons was found in the dorsal part of the contralateral RN, whereas the majority of FG-labeled neurons were located more ventrally. However, some of them were intermingled with FR-labeled neurons, and as many as one-third of FR-labeled neurons were labeled with FG. After combined injections of FR into the RFp and FG into the lower cervical spinal cord, RN neurons labeled with FG existed more ventrally than those retrogradely labeled from the upper cervical spinal cord, and less than 10% of FR-labeled neurons were labeled with FG. The present data suggest that axon collateral innervation of the RFp and the upper cervical spinal cord by single RN neurons may be responsible for coordinating head and orofacial movements.

Nigral axon terminals are in contact with parvicellular reticular neurons which project to the motor trigeminal nucleus in the rat.

We examined whether in the rat the descending fibers from the substantia nigra pars reticulata (SNr) were in contact with premotor neurons projecting to the motor trigeminal nucleus (Vm), using a combined anterograde and retrograde tracing technique. After ipsilateral injections of cholera toxin B subunit (CTb) into the Vm and biotinylated dextranamine (BDA) into the SNr, numerous CTb-labeled neurons were distributed bilaterally with slightly ipsilateral dominance in the parvicellular reticular formation (RFp), where many BDA-labeled axons with bouton-like varicosities were found bilaterally with a clear-cut ipsilateral dominance. The overlapping distribution of these labeled axons and neurons was more prominent in the rostral RFp than in the caudal RFp. Within the neuropil of the RFp, some of the BDA-labeled axons made synapses with the somata and proximal dendrites of CTb-labeled neurons. Thus, the present study demonstrated the existence of an indirect pathway from the SNr to the Vm, relayed by the RFp.

Demonstration of the corticotectobulbar pathway from the orofacial motor cortex to the parvicellular reticular formation in the rat.

We examined a corticotectobulbar pathway from the orofacial motor cortex (OfM) to the parvicellular reticular formation (RFp), where numerous premotor neurons for the orofacial motor nuclei were known to be distributed, light and electron microscopically by using a combination of anterograde and retrograde tracing techniques. After contralateral injections of biotinylated dextranamine (BDA) into the OfM and cholera toxin B subunit (CTb) into the RFp, the overlapping distribution of ipsilateral axon terminals labeled with BDA and contralateral neurons labeled with CTb was found in the lateralmost part of the superior colliculus (SC). Furthermore, contralateral injections of BDA into the OfM and wheat germ agglutinin-horseradish peroxidase (WGA-HRP) into the RFp resulted in that ipsilateral axons labeled with BDA made asymmetrical synaptic contacts with the dendrites of contralateral SC neurons labeled with WGA-HRP.

Monosynaptic and disynaptic projections from the substantia nigra pars reticulata to the parafascicular thalamic nucleus in the rat.

We examined a direct pathway and an indirect pathway via the reticular thalamic nucleus (RT) from the substantia nigra pars reticulata (SNr) to the parafascicular thalamic nucleus (PF) by using anterograde and retrograde tract tracing methods. After biotinylated dextranamine (BDA) injection into the dorsolateral part of the SNr, many labeled fibers and axon terminals were distributed in the ventral part of the RT, as well as in the ventrolateral part of the PF, bilaterally with an ipsilateral dominance. After BDA injection into the ventral part of the RT, a plexus of labeled axons was found bilaterally with an ipsilateral dominance in the ventrolateral part of the PF. After combined injections of BDA into the dorsolateral part of the SNr and cholera toxin B subunit (CTb) into the ventrolateral part of the PF on the same side, overlapping distribution of BDA-labeled fibers and CTb-labeled neurons was observed in the ventral part of the RT ipsilateral to the injection sites, where the BDA-labeled axon terminals made symmetrical synaptic contacts with soma and dendrites of the CTb-labeled neurons.

Phrenic motoneurons receive monosynaptic inputs from the Kölliker-Fuse nucleus: a light- and electron-microscopic study in the rat.

The parabrachial complex, which plays an important role in respiratory regulation, has been reported to send projection fibers to the phrenic nucleus, but the synaptic organization between the parabrachial fibers and the phrenic motoneurons has not been examined. Using anterograde and retrograde tracing methods, we found in the rat that the parabrachial fibers originating mainly from the Kölliker-Fuse nucleus (KF) terminated not only within the phrenic nucleus but also on the radial dendritic bundles of the phrenic motoneurons. It was further revealed that the KF fibers made asymmetrical synapses predominantly with dendrites and partly with somata of the phrenic motoneurons. These data suggest that output signals from the KF may exert excitatory influence directly upon the phrenic motoneurons.

Demonstration of axon collateral projections from the substantia nigra pars reticulata to the superior colliculus and the parvicellular reticular formation in the rat.

It was revealed in the rat that single neurons in the substantia nigra pars reticulata (SNr) innervated both the superior colliculus (SC) and the parvicellular reticular formation (RFp) in the pons and medulla oblongata by way of axon collaterals. After injecting Fluoro-gold into the lateral part of the SC and Fluoro-ruby into the RFp on the same side, some SNr neurons were double-labeled with both tracers. They were localized in the dorsolateral part of the caudal half of the SNr ipsilateral to the injection sites.

Organization of projections from the medial agranular cortex to the superior colliculus in the rat: a study using anterograde and retrograde tracing methods.

The organization of corticotectal projections from the medial agranular cortex (AGm), which has been considered to contain rat's frontal eye field, was examined using anterograde and retrograde tracing techniques. When biotinylated dextranamine (BDA) injections were made into the rostral part of the AGm, small numbers of BDA-labeled axons were found in the rostral two-thirds of the superior colliculus (SC) while some labeled axons were seen in the caudal one-third of the SC. These labeled axons were distributed mainly in the lateral part of the stratum griseum intermediale. On the other hand, after BDA injections into the caudal part of the AGm, moderate to dense plexuses of labeled axons were found in the rostral two-thirds of the SC while some labeled axons were seen in the caudal one-third of the SC. These labeled axons were distributed in the ventromedial and dorsolateral marginal zones of the stratum griseum intermediale as well as in the stratum griseum profundum. The corticotectal projections were largely uncrossed. After combined injections of BDA into the caudal part of the AGm on one side and cholera toxin B subunit (CTb) into the paramedian pontine reticular formation on the opposite side or into the interstitial nucleus of Cajal on the same side, the overlapping distributions of BDA-labeled axons and CTb-labeled neurons were found in the ventromedial marginal zone of the stratum griseum intermediale ipsilateral to the site of BDA injection. These results suggest that the caudal part of the AGm plays a more significant role in the oculomotor function than does the rostral part of the AGm.

Tectal projections to the parvicellular reticular formation and the upper cervical spinal cord in the rat, with special reference to axon collateral innervation.

After Phaseolus vulgaris leucoagglutinin injection into the lateral part of the superior colliculus (SC) in the rat, labeled fibers and axon terminals in the lower brainstem were distributed not only in the medial reticular formation but also in the lateral tegmental field including the parvicellular reticular formation (RFp). More caudally, in the upper cervical spinal cord labeled fibers with bouton-like varicosities were distributed mainly in laminae V, VII and VIII, with relatively sparse distribution in lamina IX. These labeled axons were found bilaterally with a clear-cut contralateral dominance. After combined injections of rhodamine-dextranamine, Fluoro-ruby (FR) into the RFp and Fluoro-gold (FG) into the upper cervical spinal cord on the same side, SC neurons labeled with FR were intermingled with those labeled with FG in the lateral part of the SC contralateral to the injection sites. In the stratum griseum intermediale, some of them were double-labeled with both tracers. Our results suggest that SC neurons innervating both the RFp and the cervical spinal cord may be involved in the coordination of head and mouth movements.

Descending projections from the superior colliculus to the reticular formation around the motor trigeminal nucleus and the parvicellular reticular formation of the medulla oblongata in the rat.

We observed by the anterograde and retrograde tracing techniques in the rat that the lateral part of the superior colliculus (SC), where the nigrotectal fibers from the dorsolateral part of the substantia nigra pars reticulata (SNr) terminated, sent projection fibers to the reticular region around the motor trigeminal nucleus (RFmt) and parvicellular reticular formation (RFp) of the medulla oblongata, where many premotor neurons for the orofacial motor nuclei were known to be distributed. The SC neurons sending their axons to the RFmt and RFp were mainly located in the stratum griseum intermedium, and additionally in the stratum griseum profundum. Our results suggest that neuronal signals conveyed through the nigro-tecto-bulbar pathway to the RFmt and RFp may exert control influences upon oral behavior.

Morphological evidence for a vestibulo-thalamo-striatal pathway via the parafascicular nucleus in the rat.

We observed by anterograde and retrograde tracing techniques that projection fibers originating from the medial vestibular nucleus (MVe) of the rat terminated in the dorsal two-thirds of the lateral part of the parafascicular thalamic nucleus (PF), where neurons sending their axons to the dorsolateral part of the striatum existed. It was further revealed that the vestibular fibers made asymmetrical synaptic contacts mainly with dendrites and additionally with soma of the striatum-projecting PF neurons. These data suggest that output signals from the MVe may be transmitted disynaptically to the striatal neurons via the PF neurons.

Accumulation of hydrocarbon in a patient with adrenoleukodystrophy.

The lipid composition of tissues from a patient with adrenoleukodystrophy was examined. Cholesterol esters of both brain and adrenal tissues contained increased proportions of fatty acids longer than C22 in agreement with previous reports. During this study, thin-layer chromatographic analysis of the neutral lipid fraction from the cerebral white matter and the adrenal gland revealed an unknown spot, which was identified as hydrocarbon of n-alkanes (CnH2n + 2, n = 16-33) by means of gas-liquid chromatography and gas chromatography/mass spectrometry. There may be a relationship between the accumulation of hydrocarbons and the change in fatty acid metabolism that is the fundamental disorder of ALD, as: (1) the fatty acids abnormally accumulated in cerebral white matter and adrenal gland coincided in chain length (i.e. C24-C26) with the main peaks of n-alkane in the same tissues, and (2) an abnormal accumulation of n-alkanes occurred in the diseased tissues where cholesterol esters with very long chain fatty acids were markedly increased.

Development of macroglial cells in the embryonic chick optic nerve.

Macroglia development in the embryonic chick optic nerve was immunohistochemically examined. The astrocytes with glial fibrillary acidic protein immunoreactivity were initially restricted to the retinal end of the optic nerve at stage 40, but had widely dispersed within the optic nerve in an out-side-in manner by stage 44. Oligodendrocytes with myelin basic protein immunoreactivity appeared at stage 38, and were widely distributed at stage 40. Electron microscopic observation confirmed the presence of intermediate filaments in the glial fibers and developing myelin at stages 40-41. The present results suggest that oligodendrocyte precursors undergo terminal differentiation slightly earlier than astrocyte precursors with respect to the expression of marker proteins.