Imaging Techniques Combined With Enzyme-Based Procedures That Facilitate the Simultaneous Imaging of Nerves, Neurons, Cartilage And Bones During Development In Gambusia

1999 ◽  
Vol 5 (S2) ◽  
pp. 1074-1075
Author(s):  
E. Rosa-Molinar
Keyword(s):  
Nervous System ◽  
Peripheral Nervous System ◽  
Imaging Techniques ◽  
Retrograde Transport ◽  
Nerve Fibers ◽  
Staining Procedure ◽  
Alizarin Red ◽  
Persistent Problem ◽  
Simultaneous Visualization ◽  
And Cartilage

A persistent problem in elucidating the anatomy of the peripheral nervous system has been an inability to stain both myelinated and unmyelinated nerve fibers. To overcome this problem, our laboratory developed two workmg protocols for reliably and differentially labeling and staining the peripheral nervous system and combined them with an enzyme clearing and staining procedure for the simultaneous visualization of bone and cartilage.One protocol uses anti-acetylated α-tubulin immunohistochemistry to follow the course, peripheral branching, and origin of the ventral spinal nerve innervating the axial musculature and a second uses anterograde and retrograde transport of selectively applied 3000 molecular weight (MW) biotin dextran amines and/or biocytin to identify specific afferent and efferent projections and their cell bodies. Both procedures can be combined with an enzyme clearing and staining procedure for the simultaneous visualization of bone (alizarin red S) and cartilage (alcian blue) in whole-mount preparations.

Multicolor Labeling for Multidimensional Microscopy: Labels to Dye for

2001 ◽  
Vol 7 (S2) ◽  
pp. 1028-1029
Author(s):  
Eduardo Rosa-Molinar ◽  
Mitch Quiron ◽  
Jeffery Smith
Keyword(s):  
Molecular Weight ◽  
Retrograde Transport ◽  
Spinal Nerve ◽  
Nerve Fibers ◽  
Molecular Probes ◽  
Staining Procedure ◽  
Alizarin Red ◽  
Motor Nerves ◽  
Axial Musculature ◽  
And Cartilage

We previously described two techniques to selectively label and/or stain myelinated and unmyelinated nerve fibers and simultaneously stain bone and cartilage. We used antiacetylated α-tubulin immunohistochemistry and anterograde and retrograde transport of selectively applied 3000 molecular weight (MW) biotin dextran amines (Molecular Probes, Inc. Eugene OR) to follow the course, peripheral branching, and origin of the ventral spinal nerve innervating the axial musculature. Those procedures were combined with an enzyme clearing and staining procedure for the simultaneous visualization of bone (alizarin red S) and cartilage (alcian blue) in whole-mount preparations. Use of the techniques enabled us to describe spatial and temporal changes of nerve fibers and the corresponding changes of the osseous elements during the transposition of the anal fin appendicular support of the Western Mosquitofish, Gambusia affinis affinis.We have extended these techniques by selective multicolor anterograde and retrograde labeling of spinal motor neuron cell bodies, their dendritic arbors, and their ventral motor nerves. We used low (3000) molecular weight (MW) fluorescent lysine-fixable dextrans (Texas Red® and fluorescein) and biotinylated lysine-fixable dextrans (biotin and fluorescein conjugated to biotin), Nissl stains, and fluorescent conjugates of cholera toxin subunit B (Molecular Probes, Inc, Eugene OR).

scholarly journals Computed Tomography in the Evaluation of Plexopathies and Proximal Neuropathies

1983 ◽  
Vol 10 (4) ◽  
pp. 244-247 ◽  
Author(s):  
John D. Stewart ◽  
Brian Schmidt ◽  
Roberto Wee
Keyword(s):  
Computed Tomography ◽  
Nervous System ◽  
Peripheral Nervous System ◽  
Imaging Techniques ◽  
Ct Scanning ◽  
Anatomical Extent ◽  
Mass Lesions ◽  
Radiological Technique

SUMMARY:We describe nine patients with plexopathies or proximal mononeuropathies due to mass lesions. In four, computed tomography (CT) was the only radiological technique to show the cause of the neuropathy. In five patients, CT either unequivocally confirmed the presence of an abnormality or was superior to other imaging techniques in showing its full anatomical extent. CT scanning is a valuable aid in the assessment of lesions of the peripheral nervous system, particularly plexopathies and mononeuropathies caused by retroperitoneal, pelvic or superior pulmonary sulcus tumors.

scholarly journals The Number of Alphaherpesvirus Particles Infecting Axons and the Axonal Protein Repertoire Determines the Outcome of Neuronal Infection

mBio ◽  
2015 ◽  
Vol 6 (2) ◽  
Author(s):  
Orkide O. Koyuncu ◽  
Ren Song ◽  
Todd M. Greco ◽  
Ileana M. Cristea ◽  
Lynn W. Enquist
Keyword(s):  
Nervous System ◽  
Peripheral Nervous System ◽  
Viral Entry ◽  
Retrograde Transport ◽  
Productive Infection ◽  
Long Distance ◽  
Viral Genomes ◽  
Neuronal Nuclei ◽  
Viral Particles ◽  
The Impact

ABSTRACTInfection by alphaherpesviruses invariably results in invasion of the peripheral nervous system (PNS) and establishment of either a latent or productive infection. Infection begins with long-distance retrograde transport of viral capsids and tegument proteins in axons toward the neuronal nuclei. Initial steps of axonal entry, retrograde transport, and replication in neuronal nuclei are poorly understood. To better understand how the mode of infection in the PNS is determined, we utilized a compartmented neuron culturing system where distal axons of PNS neurons are physically separated from cell bodies. We infected isolated axons with fluorescent-protein-tagged pseudorabies virus (PRV) particles and monitored viral entry and transport in axons and replication in cell bodies during low and high multiplicities of infection (MOIs of 0.01 to 100). We found a threshold for efficient retrograde transport in axons between MOIs of 1 and 10 and a threshold for productive infection in the neuronal cell bodies between MOIs of 1 and 0.1. Below an MOI of 0.1, the viral genomes that moved to neuronal nuclei were silenced. These genomes can be reactivated after superinfection by a nonreplicating virus, but not by a replicating virus. We further showed that viral particles at high-MOI infections compete for axonal proteins and that this competition determines the number of viral particles reaching the nuclei. Using mass spectrometry, we identified axonal proteins that are differentially regulated by PRV infection. Our results demonstrate the impact of the multiplicity of infection and the axonal milieu on the establishment of neuronal infection initiated from axons.IMPORTANCEAlphaherpesvirus genomes may remain silent in peripheral nervous system (PNS) neurons for the lives of their hosts. These genomes occasionally reactivate to produce infectious virus that can reinfect peripheral tissues and spread to other hosts. Here, we use a neuronal culture system to investigate the outcome of axonal infection using different numbers of viral particles and coinfection assays. We found that the dynamics of viral entry, transport, and replication change dramatically depending on the number of virus particles that infect axons. We demonstrate that viral genomes are silenced when the infecting particle number is low and that these genomes can be reactivated by superinfection with UV-inactivated virus, but not with replicating virus. We further show that viral invasion rapidly changes the profiles of axonal proteins and that some of these axonal proteins are rate limiting for efficient infection. Our study provides new insights into the establishment of silent versus productive alphaherpesvirus infections in the PNS.

Peripheral Nerves: Not Only Cross-sectional Area in the Era of Radiomics

2020 ◽  
Vol 24 (02) ◽  
pp. 175-180
Author(s):  
Alberto Stefano Tagliafico ◽  
Raquel Prada González ◽  
Federica Rossi ◽  
Bianca Bignotti ◽  
Carlo Martinoli
Keyword(s):  
Nervous System ◽  
Peripheral Nerve ◽  
Peripheral Nervous System ◽  
Internal Control ◽  
Peripheral Nerves ◽  
Imaging Techniques ◽  
Cross Sectional Area ◽  
Sectional Area ◽  
Musculoskeletal Radiology ◽  
Cross Sectional

AbstractThe peripheral nervous system is increasingly being investigated using medical imaging as a complement or in association with electrodiagnostics tests. The application of imaging techniques, such as ultrasound (US) and magnetic resonance imaging (MRI), allows detailed visualization of the peripheral nervous system. According to the European Society of Musculoskeletal Radiology, the use of US for nerve evaluation is strongly encouraged. In addition, the role of US is further enhanced by the wide application of US-guided techniques to diagnose or to treat peripheral nerve disorders.Standard evaluation of peripheral nerves on US usually relies on cross-sectional area evaluation with different cutoff values in the osteofibrous tunnels and outside them. In several anatomical areas, side-to-side comparison is highly recommended because it helps distinguish subtle variations by using the unaffected limb as an internal control.US is widely used to perform US-guided interventional procedures on peripheral nerves. The recent development of radiomics and machine and deep learning applied to peripheral nerves may reveal new insights beyond the capabilities of the human eye. Radiomics may have a role in expanding the diagnostic capabilities of US and MRI in the study of peripheral nerve pathology, especially when the cross-sectional area is not markedly increased.

Evidence for axonal ‘decision regions’ in the axolotl peripheral nervous system

Development ◽  
1988 ◽  
Vol 102 (4) ◽  
pp. 823-836
Author(s):  
S. Wilson ◽  
N. Holder
Keyword(s):  
Nervous System ◽  
Peripheral Nervous System ◽  
Spatial Organization ◽  
Retrograde Transport ◽  
Branch Points ◽  
Spinal Nerves ◽  
Lateral Motor Column ◽  
Decision Region ◽  
Muscle Nerve ◽  
Axial Musculature

Horseradish peroxidase (HRP) was employed to analyse the spatial organization of axons within nerves of the axolotl peripheral nervous system. HRP applications to the lateral motor column, spinal nerves and muscle nerve branches were examined after orthograde or retrograde transport. Axons change relative positions at particular limb regions, notably at the limb plexi, but also at branch points at other limb levels. Such areas of axon reorganization (termed ‘decision regions’ in line with Tosney & Landmesser (1985) J. Neurosci. 5, 2345) are interspersed by lengths of nerve in which axons run parallel to one another. A decision region is also described which involves only axons destined for axial musculature. The detailed anatomy of axon groups is discussed in terms of the likely mechanisms responsible for its formation during development. We conclude that, despite considerable variation in nerve pattern not seen in higher vertebrates, neuromuscular specificity in the axolotl limb is established largely by local pathway cues guiding axons to their appropriate targets.

scholarly journals Stereology of the Peripheral Nervous System

2019 ◽  
Vol 48 (1) ◽  
pp. 37-48 ◽  
Author(s):  
Danielle L. Brown ◽  
Michael Staup ◽  
Cynthia Swanson
Keyword(s):  
Nervous System ◽  
Peripheral Nervous System ◽  
Qualitative Evaluation ◽  
Nerve Fibers ◽  
Neuron Number ◽  
Human Eye ◽  
Additional Information ◽  
Size Number ◽  
Organ Systems ◽  
Tissue Changes

Qualitative histopathology has been the gold standard for evaluation of morphological tissue changes in all organ systems, including the peripheral nervous system. However, the human eye is not sensitive enough to detect small changes in quantity or size. Peripheral nervous system toxicity can manifest as subtle changes in neuron size, neuron number, axon size, number of myelinated or unmyelinated axons, or number of nerve fibers. Detection of these changes may be beyond the sensitivity of the human eye alone, necessitating quantitative approaches in some cases. Although 2-dimensional (2D) histomorphometry can provide additional information and is more sensitive than qualitative evaluation alone, the results are not always representative of the entire tissue and assumptions about the tissue can lead to bias, or inaccuracies, in the data. Design-based stereology provides 3D estimates of number, volume, surface area, or length, and stereological principles can be applied to peripheral nervous system tissues to obtain accurate and precise estimates, such as neuron number and size, axon number, and total intraepidermal nerve fiber length. This review describes practical stereological approaches to 3 compartments of the peripheral nervous system: ganglia, peripheral nerves, and intraepidermal nerve fibers.

Impairment Tutorial: Rating Sensory and Motor Deficits of the Upper Extremity

2000 ◽  
Vol 5 (2) ◽  
pp. 5-7
Author(s):  
Charles N. Brooks
Keyword(s):  
Nervous System ◽  
Evoked Potentials ◽  
Peripheral Nervous System ◽  
Conduction Velocity ◽  
Motor Neurons ◽  
Somatosensory Evoked Potentials ◽  
Nerve Conduction ◽  
Nerve Fibers ◽  
Nerve Conduction Studies ◽  
Electrical Potentials

Abstract The three components of electrodiagnosis useful in evaluation of the peripheral nervous system and spinal cord include electromyography (EMG), electroneurography (nerve conduction studies), and somatosensory evoked potentials. EMG examination involves introduction of a special recording needle into a muscle belly. Electrical potentials located within a few millimeters of the needle are picked up by an electrode and are transmitted from the muscle to amplifiers that filter and display results visually for the electromyographer. Three types of spontaneous activity in electrical potentials are of the greatest relevance: positive sharp waves, fibrillation potentials, and fasciculations (fasciculation potentials on the EMG result from irregular firing of motor units). Electromyography can help assess the status of nerve fibers indirectly, but the integrity of large myelinated sensory and motor neurons can be evaluated directly by nerve conduction studies (NCS), also known as electroneurography. NCS can assess motor neurons, sensory neurons, or mixed nerve trunks. Sensory nerve conduction velocity can be studied in a manner analogous to motor conduction velocity: sensory fibers can be directly stimulated, and the evoked response can be measured at the wrist and elbow. Somatosensory evoked potentials occasionally are useful as an adjunct to EMG and NCS in the diagnosis of peripheral nervous system pathology. These tests also are useful when it is unclear whether an individual has a true radiculopathy.

Electrodiagnosis of the Peripheral Nervous System: An Introduction

2014 ◽  
Vol 19 (3) ◽  
pp. 10-14
Author(s):  
Richard T. Katz
Keyword(s):  
Nervous System ◽  
Evoked Potentials ◽  
Spontaneous Activity ◽  
Peripheral Nervous System ◽  
Motor Neurons ◽  
Somatosensory Evoked Potentials ◽  
Nerve Conduction ◽  
Nerve Fibers ◽  
Nerve Conduction Studies ◽  
Mixed Nerve

Abstract This article is an introduction to electrodiagnosis of the peripheral nervous system, including electromyography, electroneurography (nerve conduction studies), and somatosensory evoked potentials. Electromyography involves the introduction of a special recording needle into a muscle body in search of spontaneous activity (electrical potentials that occur while the muscle is at rest). Three types of spontaneous activity are of greatest relevance: positive sharp waves, fibrillation potentials, and fasciculations. Electromyography can help assess the status of nerve fibers indirectly, but the integrity of large myelinated sensory and motor neurons can be evaluated directly by nerve conduction studies (NCS), also known as electroneurography. NCS involves the introduction of an electrical stimulus, either by surface electrode or needle, and recording an evoked response. NCS can assess motor neurons, sensory neurons, or mixed nerve trunks, depending on the strategy employed. Somatosensory evoked potentials (SSEP) sometimes are useful as an adjunct to EMG and NCS in the diagnosis of peripheral nervous system pathology and are obtained by stimulating a peripheral mixed nerve at a frequency of approximately 2-5 Hz. Several manufacturers have created automated, hand-held units for performing nerve conduction studies, and neuromuscular ultrasound is noninvasive and painless, and ultrasound of nerve entrapment has identified nerve enlargement just proximal to the site of entrapment. Physicians should know or learn the qualifications of the physician to whom they refer their patients for electrodiagnostic assessment.

scholarly journals Dynamic ubiquitination drives herpesvirus neuroinvasion

2015 ◽  
Vol 112 (41) ◽  
pp. 12818-12823 ◽  
Author(s):  
Nicholas J. Huffmaster ◽  
Patricia J. Sollars ◽  
Alexsia L. Richards ◽  
Gary E. Pickard ◽  
Gregory A. Smith
Keyword(s):  
Nervous System ◽  
Peripheral Nervous System ◽  
Nerve Fibers ◽  
Peripheral Tissue ◽  
Nerve Endings ◽  
Healthy Individuals ◽  
Virus Particles

Neuroinvasive herpesviruses display a remarkable propensity to enter the nervous system of healthy individuals in the absence of obvious trauma at the site of inoculation. We document a repurposing of cellular ubiquitin during infection to switch the virus between two invasive states. The states act sequentially to defeat consecutive host barriers of the peripheral nervous system and together promote the potent neuroinvasive phenotype. The first state directs virus access to nerve endings in peripheral tissue, whereas the second delivers virus particles within nerve fibers to the neural ganglia. Mutant viruses locked in either state remain competent to overcome the corresponding barrier but fail to invade the nervous system. The herpesvirus “ubiquitin switch” may explain the unusual ability of these viruses to routinely enter the nervous system and, as a consequence, their prevalence in human and veterinary hosts.

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