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PubMed1997

Expression of the cell adhesion molecule axonin-1 in neuromeres of the chicken diencephalon.

Axonin-1/TAG-1, a member of the immunoglobulin (Ig) superfamily of adhesion molecules, has been shown to be selectively expressed by a subset of neurons and fiber tracts in the developing nervous system of vertebrates. Axonin-1/TAG-1 is thought to play a role in the outgrowth, guidance, and fasciculation of neurites. In the present study, we map the expression of axonin-1 in the diencephalon of the chicken brain at early and intermediate stages of development [2-8 days of incubation; embryonic day (E)2-E8] by immunohistochemical methods. Results show that axonin-1 is first expressed at about E2.5 by postmitotic neurons scattered throughout most of the diencephalon. During the neuromeric stage of brain development (about E3-E5), axonin-1+ nerve cell bodies are predominantly found in two neuromeric subdivisions: 1) in the alar plate of the precommissural pretectum and dorsal thalamus and 2) in the posterior preoptic region of the hypothalamus. The axonin-1+ fiber bundles emerging from these areas grow across segmental boundaries. For example, axonin-1+ neurites originating in the dorsal thalamus cross the zona limitans intrathalamica at a right angle to project to the striatum. Later, the axonin-1+ neuromere areas give rise to particular axonin-1+ gray and white matter structures. Most of these structures correspond to the structures described to express TAG-1 in rodents. In conclusion, axonin-1 can be used as a marker to study aspects of the transition from the early neuromeric structure to the mature anatomy of the chicken brain.

Animals↗
PubMed1997

Development of differential preganglionic projections to pre- and paravertebral sympathetic ganglia.

Sympathetic preganglionic axons project to spatially distinct targets in the periphery. A precise topographic pattern exists within the thoracic preganglionic cell column relative to the direction of axonal projections within the sympathetic chain. In this study, the time course and pattern of axonal outgrowth from different populations of preganglionic neurons in the chicken embryo is examined in detail to clarify the origin of the topography in this system. Projections to prevertebral targets are established by development of the splanchnic nerves by stage 25, well after the earliest somatic motor projections at stage 19 but at least two stages before the reported onset of paravertebral projections. Further, preganglionic axons that project rostrally into the sympathetic chain may do so earlier than those that project caudally in the chain. The separation of preganglionic axons into prevertebral, rostral paravertebral or caudal paravertebral directions occurs at a common site in the ventral mesenchyme, established by the initial ventromedial projection of the splanchnic nerves. Analysis of the axonal trajectories of rostrally and caudally projecting cells reveals that preganglionic axons are not selectively fasciculated before their point of separation at the sympathetic chain. The patterning of the preganglionic cell column is specified before the establishment of functional connections within the chain, indicating that target contact is not a determinant of the segmental pattern. We suggest that the differential outgrowth of preganglionic axons to peripheral targets is determined by the unique identities of underlying subpopulations of preganglionic axons.

Acetylcholinesterase↗
PubMed1997

Development of phrenic motoneuron morphology in the fetal rat.

This study examined the morphological changes that a homogeneous mammalian spinal motoneuron population undergoes during foetal development. Retrograde labelling of the phrenic nerve with the carbocyanine dye, DiI, was used to visualise developmental changes in phrenic motoneuron morphology within the cervical spinal cord of perinatal rats from embryonic day (E) 13.5 to birth (ca. E21). Groups of intimately associated phrenic somata had migrated into the ventromedial region of cervical segments C3-C6 by E14. This migration was followed by their progressive compaction into a tightly aligned column by E18. During this period, close contact was maintained between phrenic somata throughout the motor pool, suggestive of the presence of gap junctions. From E15 to E18, extensive dendritic arborisations fanned out dorsolaterally and ventromedially into the white matter and the floor plate. By E19, however, dendritic fasciculation and retraction and the extension of newly formed rostrocaudally projecting dendrites had resulted in the approximation of the dendritic morphology observed at birth. These data demonstrate that morphological maturation of phrenic motoneurons occurs subsequently to the onset of functional recruitment and the arrival of central processes of dorsal root ganglion neurons within the ventral horn (ca. E17). By birth, a number of immature features remain, including a larger proportion of neurites that project into the white matter and into the floor plate, the presence of growth cones on a number of dendrites, and close contact between populations of contralaterally derived dendrites.

Animals↗
PubMed2000

Remodeling of the leg sensory system during metamorphosis of the hawkmoth, Manduca sexta.

The adult legs of the hawkmoth Manduca sexta are supplied by a diverse array of sensory organs and associated neurons (Kent and Griffin [1990] Cell Tissue Res. 259:209-223) that differ from those in the larval legs. In the present study, a combination of nerve-tracing techniques [biocytin, 1,1;-dioctadecyl-3,3,3;, 3;-tetramethyl-indocarbocyanine perchlorate (DiI)], birth date labeling (5-bromodeoxyuridine), confocal microscopy, and electrophysiology were used to describe the remodeling of the prothoracic leg sensory system. Four primary sensory branches carry the axons of all of the sensory neurons in the larval leg. At the onset of metamorphosis, the imaginal leg epidermis develops underneath the larval cuticle and encircles the sensory neurons, thus separating them from their target-organs. Most of the larval neurons degenerate during the larval-to-pupal transition and are replaced by new-adult sensory neurons that are born and differentiate in the pupa. Six sensory neurons that supply hair sensilla in the larval leg, together with 13 femoral and tibial chordotonal organ neurons, persist into the developing adult leg to serve similar functions. Early in the pupal stage, electrical activity can be recorded from these neurons despite the absence of target sensory structures. During the differentiation of the adult sensory system, the axons of the new-adult sensory neurons contact and fasciculate with the axons of the persistent neurons. Thus, five of the primary sensory branches of the adult leg are built on the preexisting larval sensory trajectories. Two sensory branches, however, are established de novo by the axons of specific adult sensory neurons.

Aging↗
PubMed1999

Isolation, primary structure characterization and identification of the glycosylation pattern of recombinant goldfish neurolin, a neuronal cell adhesion protein.

Neurolin is a growth-associated cell surface glycoprotein from goldfish and zebra fish which has been shown to be involved in axonal path-finding in the goldfish retina and suggested to function as a receptor for axon guidance molecules. Being a member of the immunoglobulin superfamily of cell adhesion proteins, neurolin consists of five N-terminal extracellular immunoglobulin (Ig)-like domains, a transmembrane and a short cytoplasmatic domain. Repeated injections of polyclonal Fab fragments against neurolin and of monoclonal antibodies against either Ig domains cause path-finding errors and disturbance of axonal fasciculation. In order to obtain a complete structural characterization and a molecular basis for structure-function determination, recombinant neurolin with the complete extracellular part but lacking the transmembrane and cytoplasmatic domain was expressed in Chinese hamster ovary (CHO) cells (CHO-neurolin). The isolation of CHO-neurolin was carried out by Ni-affinity chromatography and subsequent high-performance liquid chromatography (HPLC). An exact molecular mass determination was obtained by matrix-assisted laser desorption/ionization mass spectrometry (MALDI/MS) and revealed 60.9 kDa, which suggested that approximately 10 kDa are due to glycosylation. The predicted molecular mass is 51.5 kDa, whereas sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) yielded an apparent molecular mass of 72 kDa. Gel shift assays using SDS-PAGE and Western blot analysis with anti-neurolin antibodies provided consistent molecular mass data. The complete primary structure and N-glycosylation patterns were identified using specific lectin assays, MALDI/MS peptide mapping analysis by proteolytic and in-gel digestion, electrospray ionization MS and MALDI/MS in combination with specific glycosidase degradation. HPLC isolation of glycosylated peptide fragments and MS after selective deglycosylation revealed heterogeneous glycosylations at all five N-glycosylation consensus sites. All attached N-glycans are of the complex type and show a mainly biantennary structure; they are fucosylated with alpha(2,3)-terminal neuraminic acid. These data serve as a first detailed model to characterize the molecular recognition structures exhibited by the extracellular domains.

Activated-Leukocyte Cell Adhesion Molecule↗
PubMed2000

Kaposi sarcoma of major salivary gland origin: A clinicopathologic series of six cases.

BACKGROUND: Kaposi sarcoma (KS), one of the defining tumors of acquired immune deficiency syndrome (AIDS), is rarely identified in the major salivary glands. To the authors' knowledge, no previous published series has evaluated the clinicopathologic aspects of this tumor in major salivary glands. METHODS: Six cases of salivary gland KS, diagnosed between 1970 and 1998, were retrieved from the files of the Oral and Maxillofacial Pathology Registry of the Armed Forces Institute of Pathology. Histologic features were reviewed and special stains, immunohistochemical studies, and in situ hybridization were performed (n = 4). Patient follow-up data were obtained. RESULTS: The patients included 6 men ages 20-73 years (average, 53.0 years). Patients presented clinically with a mass in the submandibular (n = 4) or parotid (n = 2) gland region. Symptoms were present for a mean of 13.7 months. The tumors measured 1-4 cm (average, 2.5 cm) in greatest dimension. Histologically, the tumors exhibited the usual features of KS: a spindle cell vascular proliferation arranged in fasciculated bundles, variable nuclear pleomorphism, mitotic figures, extravasated erythrocytes, and hyaline globules. Five patients were serologically positive for human immunodeficiency virus (HIV) (three homosexual males, one infected by a contaminated blood transfusion, and one with an unknown risk factor). Human herpesvirus-8 (HHV-8) was present in all cases tested (n = 4). Patients were treated with surgical excision (n = 6), followed by chemotherapy (n = 1) for the single patient with other foci of KS (rectal). Three patients died of AIDS-related infectious complications and one of congestive heart failure, whereas the remaining patients are alive with AIDS but free of salivary gland KS. CONCLUSIONS: Salivary gland enlargement is frequently identified in HIV positive or AIDS patients. Although rare, it is important to consider KS in the differential diagnosis of other AIDS-related salivary gland manifestations (infections and tumors).

AIDS-Related Opportunistic Infections↗
PubMed1998

Fine-needle aspiration cytology of malignant retroperitoneal paraganglioma.

The cytologic findings in a fine-needle aspiration sample are described from a large retroperitoneal mass in a 56-yr-old male. The aspiration was performed under CT guidance using a 22-gauge needle, maintaining negative pressure. For cytologic study, on-site smears were prepared and stained by the Papanicolaou method. Additionally, cytospin, filter preparations, cell blocks from the aspirate, histology, and electron microscopy of the tumor tissue were performed. Also, immunodiagnostic staining for neuron-specific enolase (NSE), chromogranin, S-100 protein, vimentin, HMB45, cytokeratin, and Grimelius preparation was performed on cytologic and histologic material. The cytologic material was characterized by cords, suggestive acinar structures, and small follicle-like clusters of cells, while cell blocks from the aspirate showed oval or spindle-shaped nuclei with a somewhat fasciculated appearance. However, sections from the tumor tissue showed an organoid "zellballen" pattern. The nuclei were round, oval, spindly, and epithelioid, with moderate to scanty cytoplasm, smooth nuclear membranes, fine, evenly dispersed chromatin, slight hyperchromasia, and mild pleomorphism. No intranuclear vacuoles were seen. Positive immunohistochemical staining for NSE, chromogranin, and Grimelius preparation was noted in the tumor cells, while staining for vimentin, HMB45, and cytokeratin was negative. Electron microscopy of the tumor tissue revealed the presence of variable numbers of round, membrane-bound, electron-dense neurosecretory granules. The cytohistologic and ultrastructural findings are presented, as well as the results of immunodiagnostic staining which helped in the diagnosis of retroperitoneal paraganglioma, an infrequently reported tumor.

Biopsy, Needle↗
PubMed1997

Tyrosine phosphorylation of L1 family adhesion molecules: implication of the Eph kinase Cek5.

The L1 family comprises transmembrane cell adhesion molecules of the immunoglobulin superfamily that play an important role in neuronal migration and axon outgrowth, fasciculation, and myelination. Consistent with a crucial role in developmental processes, mutations in L1 cause severe brain malformations. Although L1 activates intracellular signaling pathways, little is known about the membrane proximal events of L1 signaling. The cytoplasmic domains of L1 family proteins contain several conserved tyrosine residues that are potential targets for receptor tyrosine kinases. Here, we report that the L1 family protein Ng-CAM is phosphorylated on tyrosine in embryonic day 13 chicken retina. This is the first demonstration of in vivo tyrosine phosphorylation of an L1-like molecule. Because chicken embryo kinase 5 (Cek5) is a receptor tyrosine kinase expressed in neuronal processes and activated in the chicken embryonic retina, we have analyzed the possible role of Cek5 in L1 phosphorylation. The rat glioblastoma cell line B28 was stably transfected with human L1. Additional transient transfection with Cek5 cDNA led to expression of Cek5 in its tyrosine-phosphorylated, activated form. Biochemical analysis revealed that L1 is phosphorylated on tyrosine in Cek5-transfected cells but not in control transfectants. Furthermore, direct phosphorylation of the L1 cytoplasmic domain by Cek5 was demonstrated in an in vitro kinase assay. Tyrosine phosphorylation may represent a novel mechanism of signal cascade initiation through L1.

Amino Acid Sequence↗
PubMed1997

Neural cell adhesion molecule L1: signaling pathways and growth cone motility.

The neural cell adhesion molecule L1 plays a key role in nervous system development including neuronal migration, neurite growth, and axonal fasciculation. L1 is expressed on most developing axons, and homophilic binding of L1 molecules on adjacent axons is likely to play a key role in axon extension. It is now well documented that a number of second-messenger systems are involved in L1-stimulated neurite growth in vitro. However, it is unclear how L1 homophilic or heterophilic binding trigger signals that regulate the mechanical forces that produce axon extension. In this report, we will review recent advances in understanding L1-associated signals, L1 interactions with the cytoskeleton, and the molecular mechanisms underlying growth cone motility.

Animals↗
PubMed1996

AAEM case report #30: multifocal motor neuropathy.

A 73-year-old man with a 16-year history of fasciculations and 15 years of weakness in his right arm was diagnosed with focal motor neuron disease. After 10 years of purely motor symptoms, he developed mild parasthesias although his sensory examination remained normal. Reflexes were reduced or absent in the weak muscles but were normal elsewhere. Nerve conduction was studied in nerves innervating weak muscles and showed severe motor conduction block. Sensory nerve conduction studies were minimally abnormal, showing reduced amplitudes with normal velocities. Based on the clinical picture and the presence of severe motor conduction block, the patient was diagnosed as multifocal motor neuropathy. Treatment with high-dose intravenous immunoglobulin was given with significant improvement in strength and partial resolution of the conduction block. As this case demonstrates, this treatable disorder may occasionally be mistaken for motor neuron disease although the resemblance is only superficial, and it should never be mistaken for amyotrophic lateral sclerosis. Multifocal motor neuropathy is an inflammatory, demyelinating neuropathy which, like chronic inflammatory demyelinating polyneuropathy (CIDP), is probably immune-mediated. It differs from typical CIDP by virtue of a marked predilection for motor axons, a strikingly restricted distribution, and a protracted course. Treatment with high-dose intravenous immunoglobulin is frequently helpful, but other forms of immune manipulation are less effective.

Demyelinating Diseases↗
PubMed1999

Central motor conduction time in patients with multifocal motor conduction block.

The finding of conduction block (CB) within short consecutive segments along a motor nerve is a key feature of multifocal motor neuropathy (MMN). Despite their different pathogenesis, this may be the only clinical difference between some cases of MMN and the pure spinal muscular atrophy form of motor neuron disease (MND). In 12 patients with distal atrophy and fasciculations and electrophysiological evidence of CBs in the upper limbs, we measured the peripheral and central motor conduction times (PMCT and CMCT) to hand muscles. We reasoned that patients with MMN should show an abnormally prolonged PMCT with normal CMCT, whereas an increased CMCT would suggest MND. All patients had delayed F-wave latency and increased PMCT. Three patients had increased CMCT. Follow-up showed little clinical and electrophysiological change in 7 of the 9 patients with normal CMCT, and a progressive motor deficit leading ultimately to death in 1 of the 3 patients with increased CMCT. This patient's electrophysiological follow-up showed a significant decrement of the compound motor action potential to both proximal and distal stimulation points, with disappearance of earlier CBs. Autopsy revealed loss of anterior horn cells and axons of the ventral root, and degeneration of large myelinated fibers. We conclude that determining the CMCT may help in differentiating MND from MMN. Persistence of a stable clinical picture over a span of at least 1 year and lack of electrophysiological signs of involvement of upper motor neurons should both be required before establishing the diagnosis of MMN even with electrophysiological evidence of CB.

Adult↗
PubMed1999

Distinguishing clinical and electrodiagnostic features of X-linked bulbospinal neuronopathy.

X-linked bulbospinal neuronopathy (XLBSN) or Kennedys disease is a rare inherited neuromuscular disease characterized by adult-onset muscle weakness, usually in a limb-girdle distribution. It is frequently misdiagnosed despite a distinctive clinical presentation, usually due to the absence of a clear family history, and perhaps also due to failure of recognition. Accurate diagnosis is crucial for genetic counseling purposes and because alternative diagnoses usually carry a poorer prognosis. We evaluated 4 patients with XLBSN and one symptomatic female heterozygote patient. Based on our clinical observations in these patients and a systematic review of previously reported cases, the following clinical and electrophysiologic features when present in the setting of adult-onset muscle weakness, are strongly suggestive of the disorder: 1) facial weakness, 2) facial twitching or fasciculations, 3) tongue weakness and atrophy, 4) postural hand tremor, 5) hypo- or areflexia, and 6) absent or low-amplitude sensory nerve action potentials despite clinically normal sensation. We also hypothesize regarding the possibility of partial expression of the abnormal XLBSN gene in a symptomatic heterozygote female patient.

Action Potentials↗
PubMed2000

Chronic motor axonal neuropathy associated with antibodies monospecific for N-acetylgalactosaminyl GD1a.

We report on three patients with chronic motor neuropathy who had elevated titers of immunoglobulin (Ig)G antibodies against N-acetylgalactosaminyl GD1a (GalNAc-GD1a) and normal titers of antibodies against other gangliosides. Presenting with progressive muscular atrophy, fasciculations, and no sensory deficits, the patients had been diagnosed to have motor neuron disease. Electrodiagnostic features were predominantly axonal. Two patients clinically improved after intravenous Ig infusion and cyclophosphamide therapy. Increased titers of IgM antibodies to GalNAc-GD1a were also found in two of 15 patients with multifocal motor neuropathy with conduction block but were associated with concomitant rise of anti-GM1 antibodies. These three cases represent a chronic motor axonal neuropathy in which antibody testing for a minor ganglioside was helpful for instituting therapy.

Action Potentials↗
PubMed1996

Elucidation of the molecular actions of NCAM and structurally related cell adhesion molecules.

The Neural Cell Adhesion Molecule (NCAM) is a founder member of a large family of cell surface glycoproteins that share structural motifs related to immunoglobulin and fibronectin type III (FN III) domains [Walsh and Doherty (1991) (Fig. 1). These glycoproteins have been grouped based on the respective number of each type of domain. In vertebrates members of this family of glycoproteins include L1/NILE, NgCAM, axonin-1/TAG-1, and Thy-1 as well as NCAM. In addition structural homologs of NCAM and L1 have been identified in Drosophila and Grasshoppers [Walsh and Doherty (1991)]. These insect homologs are called fasciclins and a series of mutants corresponding to these aspects of synaptic plasticity [Mayford et al. (1992) Science 256:638-644]. In vertebrates all of these glycoproteins are expressed in the developing nervous system where they have been identified as candidate molecules for mediating axon outgrowth, fasciculation, regeneration, and target recognition. In addition, NCAM is expressed in a number of different tissues and cell types. For example, NCAM is expressed in a dynamic pattern in developing and regenerating adult muscle. In this review we aim to describe important aspects of the role of these CAMS in development of the nervous system, including the neuromuscular junction. Furthermore, we will explore the prospective use of molecular biology, cell biology, and molecular genetic techniques, such as transgenic mice, to understand the role and molecular action of this family of cell adhesion molecules in vivo.

Amino Acid Sequence↗
PubMed1996

Developmentally regulated expression of a cell surface protein, neuropilin, in the mouse nervous system.

Neuropilin (previously A5) is a cell surface glycoprotein that was originally identified in Xenopus tadpole nervous tissues. In Xenopus, neuropilin is expressed on both the presynaptic and postsynaptic elements in the visual and general somatic sensory systems, suggesting a role in neuronal cell recognition. In this study, we identified a mouse homologue of neuropilin and examined its expression in developing mouse nervous tissues. cDNA cloning and sequencing revealed that the primary structure of the mouse neuropilin was highly similar to that of Xenopus and that the extracellular segment of the molecule possessed several motifs that were expected to be involved in cell-cell interaction. Immunohistochemistry and in situ hybridization analyses in mice indicated that the expression of neuropilin was restricted to particular neuron circuits. Neuropilin protein was localized on axons but not on the somata of neurons. The expression of neuropilin persisted through the time when axons were actively growing to form neuronal connections. These observations suggest that neuropilin is involved in growth, fasciculation, and targeting for a particular groups of axons.

Amino Acid Sequence↗
PubMed1997

Activity-dependent regulation of N-cadherin in DRG neurons: differential regulation of N-cadherin, NCAM, and L1 by distinct patterns of action potentials.

Cell adhesion molecule (CAM) expression is highly regulated during nervous system development to control cell migration, neurite outgrowth, fasciculation, and synaptogenesis. Using electrical stimulation of mouse dorsal root ganglion (DRG) neurons in cell culture, this work shows that N-cadherin expression is regulated by neuronal firing, and that expression of different CAMs is regulated by distinct patterns of neural impulses. N-cadherin was down-regulated by 0.1 or 1 Hz stimulation, but NCAM mRNA and protein levels were not altered by stimulation. L1 was down-regulated by 0.1 Hz stimulation, but not by 0.3 Hz, 1 Hz, or pulsed stimulation. N-cadherin expression was lowered with faster kinetics than L1 (1 vs. 5 days), and L1 mRNA returned to higher levels after terminating the stimulus. The RSLE splice variant of L1 was not regulated by action potential stimulation, and activity-dependent influences on L1 expression were blocked by target-derived influences. The results are consistent with changes in firing pattern accompanying DRG development and suggest that functional activity can influence distinct developmental processes by regulating the relative abundance of different CAMs.

Action Potentials↗
PubMed1998

Effects of ion channel activity on development of dorsal root ganglion neurons.

Studies of mouse dorsal root ganglion neurons in vitro demonstrate that ion channel function and regulation can influence a wide range of developmental processes. The work suggests that much as exposure to different trophic factors, the pattern of impulse activity a neuron experiences can have significant structural and functional effects during development. Studies concerning effects of ion channel activity on growth cone motility, axon fasciculation, synaptic plasticity, myelination, and intracellular signaling pathways regulating gene expression are presented in the context of changes in endogenous firing patterns during development.

Action Potentials↗
PubMed2000

Conserved developmental algorithms during thalamocortical circuit formation in mammals and reptiles.

The general patterns of early thalamocortical development follow a similar sequence in all mammals. Thalamocortical projections descend through the ventral thalamus, advance in the internal capsule amongst cells which already possess dorsal thalamic projections, then reach the cerebral cortex by associating with subplate cells and their early corticofugal projections. Initially, the thalamic projections pause in the internal capsule and subplate layer. The interactions of the thalamocortical projections with the early generated, largely transient cells of the subplate, marginal zone, internal capsule and ventral thalamus are believed to play a crucial role in the organized deployment of thalamic projections and establishing a functional cortical architecture. Selective fasciculation, contact guidance and release of neurotrophic factors are thought to play roles in the development of thalamocortical projections. These ideas are obtaining support from recent work on reeler and other strains of mice. The evolutionary origin of these largely transient cells and the overlying logic of early developmental steps are not understood. The behaviour of the thalamocortical and corticothalamic projections at the corticostriatal junction is particularly puzzling. The comparison of early forebrain development in mammals and reptiles is beginning to reveal highly conserved cellular and molecular interactions during early thalamocortical development and to reveal homologies between telencephalic subdivisions.

Algorithms↗