Efficacy of seven retrograde tracers, compared in multiple-labelling studies of feline motoneurones

Richmond, F.J.R.; Gladdy, Rebecca A.; Creasy, J.L.; Kitamura, Seiichiro; Smits, Emely; Thomson, D. B.

doi:10.1016/0165-0270(94)90142-2

Cited by 116 publications

(98 citation statements)

References 40 publications

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“…The phrenic nerve was cut without opening the pleural cavity, and a piece of hemostatic sponge (Spongostan; Ferrosan, Soeborg, Denmark) 1 ϫ 1 ϫ 1 mm in size was soaked in 10% Fluoro-ruby (Chemicon) in Ringer's solution with 2% DMSO, and applied to the proximal stump. Labeling by dextran tracers is restricted to cut axons (Richmond et al, 1994), and care was taken not to injure other branches of the cervical plexus or muscles innervated by it. The wound was closed in two layers with 9-0 and 6-0 sutures.…”

Section: Methodsmentioning

confidence: 99%

Progressive Motor Neuronopathy: A Critical Role of the Tubulin Chaperone TBCE in Axonal Tubulin Routing from the Golgi Apparatus

Schaefer¹,

Schmalbruch²,

Buhler³

et al. 2007

J. Neurosci.

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Axonal degeneration represents one of the earliest pathological features in motor neuron diseases. We here studied the underlying molecular mechanisms in progressive motor neuronopathy ( pmn) mice mutated in the tubulin-specific chaperone TBCE. We demonstrate that TBCE is a peripheral membrane-associated protein that accumulates at the Golgi apparatus. In pmn mice, TBCE is destabilized and disappears from the Golgi apparatus of motor neurons, and microtubules are lost in distal axons. The axonal microtubule loss proceeds retrogradely in parallel with the axonal dying back process. These degenerative changes are inhibited in a dose-dependent manner by transgenic TBCE complementation that restores TBCE expression at the Golgi apparatus. In cultured motor neurons, the pmn mutation, interference RNA-mediated TBCE depletion, and brefeldin A-mediated Golgi disruption all compromise axonal tubulin routing. We conclude that motor axons critically depend on axonal tubulin routing from the Golgi apparatus, a process that involves TBCE and possibly other tubulin chaperones.

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Section: Methodsmentioning

confidence: 99%

Progressive Motor Neuronopathy: A Critical Role of the Tubulin Chaperone TBCE in Axonal Tubulin Routing from the Golgi Apparatus

Schaefer¹,

Schmalbruch²,

Buhler³

et al. 2007

J. Neurosci.

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“…Despite the abundance of commercially available fluorescent retrograde neuronal tracers, the choice of appropriate tracers for this application is frequently difficult (Richmond et al, 1994;Novikova et al, 1997;Vercelli et al, 2000). In order to choose a tracer which is effective for labeling motoneurons, either before or after reinnervation, an understanding of the persistency of the tracer and number of labeled cells is required.…”

Section: Double-labeled Cells Would Then Identify Native Neurons Thatmentioning

confidence: 99%

A sequential double labeling technique for studying changes in motoneuronal projections to muscle following nerve injury and reinnervation

Katada¹,

Vos²,

Swelstad³

et al. 2006

Journal of Neuroscience Methods

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Abstract:The purpose of this study was to develop an anatomical technique that could directly demonstrate the motoneuron projections to the muscle both before injury and again following reinnervation. Investigation focused on the identification of a long-term retrograde fluorescent tracer that would label original motoneurons and persist long enough for reinnervating motoneurons to become labeled by a second fluorescent tracer. True Blue (TB) was evaluated as a potential long-term tracer, Fluoro-ruby (FR) and Fluoro-emerald (FE) were tested as potential short-term tracers in 45 adult Sprague-Dawley rats. In the initial phase of the study, TB was injected into the tibialis anterior (TA) muscle in 16 rats and sacrificed 1 week to 6 months later, to study its persistence. During the second stage, a short-term tracer was injected into the TA muscles bilaterally in 15 rats with survival time ranging from 4-28 days. Sequential double labeling was subsequently performed using the combination of TB and FR in 14 rats. The number and brightness of TB cells did not change over 6 months time, a period sufficient for complete reinnervation.FR and FE showed maximum labeling of motoneurons at 1 week after tracer application.In the double labeling study, we could easily distinguish double-labeled cells from those labeled only by TB or FR. These results suggest that sequential double labeling of TB and FR is a valuable method for long-term muscle reinnervation studies.

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“…Injection of a dye solution into the muscle (Kristensson, 1970;Richmond et al, 1994) . Pressure injection of a dye solution into the proximal nerve stump followed by ligature of the nerve stump to prevent dye leakage (Schmid et al, 1983).…”

Section: Retrograde Tracing Of Spinal Cord Motor Neuronesmentioning

confidence: 99%

“…Fluorescent dyes with dierent excitation wavelengths and physico-chemical properties, and which are known to be non-toxic, are selected as local tracers from the site of the sciatic nerve lesion. The carbocyanine dyes, DiI, DiO (Honig and Hume, 1989;Novikova et al, 1997;Richmond et al, 1994), and DiAsp (Thanos et al, 1994 for review) and FG (Schmued and Fallon, 1986) may be used as tracers. Small crystals of the dye are placed onto the proximal nerve stump and held in place with resorbable gelfoam.…”

Section: Retrograde Tracing Of Spinal Cord Motor Neuronesmentioning

confidence: 99%

Current concepts in neuroanatomical tracing

Köbbert

Apps

Bechmann

et al. 2000

Progress in Neurobiology

377

335

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The development of new axonal tract tracing and cell labelling methods has revolutionised neurobiology in the last 30 years. The aim of this review is to consider some of the key methods of neuroanatomical tracing that are currently in use and have proved invaluable in charting the complex interconnections of the central nervous system. The review begins with a short overview of the most frequently used tracers, including enzymes, peptides, biocytin, latex beads, plant lectins and the everincreasing number of¯uorescent dyes. This is followed by a more detailed consideration of both well established and more recently introduced neuroanatomical tracing methods. Technical aspects of the application, uptake mechanisms, intracellular transport of tracers, and the problems of subsequent signal detection, are also discussed. The methods that are presented and discussed in detail include: (1) anterograde and retrograde neuroanatomical labelling with¯uorescent dyes in vivo, (2) labelling of post mortem tissue, (3) developmental studies, (4) transcellular tracing (phagocytosis-dependent staining of glial cells), (5) electrophysiological mapping combined with neuronal tract tracing, and (6) simultaneous detection of more than one axonal tracer. (7) Versatile protocols for three-colour labelling have been developed to study complex patterns of connections. It is envisaged that this review will be used to guide the readers in their selection of the most appropriate techniques to apply to their own particular area of interest. 7

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Efficacy of seven retrograde tracers, compared in multiple-labelling studies of feline motoneurones

Cited by 116 publications

References 40 publications

Progressive Motor Neuronopathy: A Critical Role of the Tubulin Chaperone TBCE in Axonal Tubulin Routing from the Golgi Apparatus

Progressive Motor Neuronopathy: A Critical Role of the Tubulin Chaperone TBCE in Axonal Tubulin Routing from the Golgi Apparatus

A sequential double labeling technique for studying changes in motoneuronal projections to muscle following nerve injury and reinnervation

Current concepts in neuroanatomical tracing

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