Tetramethyl benzidine for horseradish peroxidase neurohistochemistry: a non-carcinogenic blue reaction product with superior sensitivity for visualizing neural afferents and efferents.

Mesulam, Marek-Marsel

doi:10.1177/26.2.24068

Cited by 4,268 publications

(773 citation statements)

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“…Following their removal from the skull and postfixation for 2-4 h in the same fixative, the brainstems were immersed overnight at 4°C in 20% sucrose in 0.12 M phosphate buffer, pH 7.4. Forty/~m thick transverse frozen sections were cut containing the cochlear nuclei (CN), superior olivary complex (SOC), nuclei of the lateral lemniscus (NLL) and inferior colliculus (IC) and reacted with 3,3 '-5,5 '-tetramethylbenzidine (TMB) to visualize the peroxidase (Mesulam, 1978). The reacted sections were then mounted and counterstained with neutral red (Mesulam, 1978).…”

Section: Methodsmentioning

confidence: 99%

Connections between the cochlear nuclei in guinea pig

et al. 1992

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This study provides a detailed analysis of the appearances and distributions of neurons projecting from one cochlear nucleus to the other. Injections of wheatgerm agglutinin conjugated to horseradish peroxidase were made into ventral or dorsal cochlear nucleus of the guinea pig. Retrogradely labeled cells in the opposite cochlear nucleus were examined and quantified. Three major categories of labeled cells were discerned on the basis of their soma shape: elongate, round-to-oval, and polygonal. All injections resulted in widespread labeling of cells in all of these categories, but especially round-to-oval cells, in the opposite ventral cochlear nucleus and sparse labeling in the dorsal cochlear nucleus. The results suggest that there is a significant cochlear nucleus commissural projection involving heterogeneous cell types which could have diverse functions in binaural auditory signal processing.

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Section: Methodsmentioning

confidence: 99%

Connections between the cochlear nuclei in guinea pig

et al. 1992

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“…The craniomedial aspect of the left shoulder was approached, 15 the musculocutaneous, suprascapular and subscapular nerves were exposed, cleared from surrounding tissues and transected. HRP (Sigma, type VI) was applied to the proximal stump of the nerves for 1 h. 16 The exposed area was then washed out with saline and the surgical wound was closed in layers.…”

Section: Postoperative Periodmentioning

confidence: 99%

Lateral approach of the dog brachial plexus for ventral root reimplantation

et al. 1998

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A lateral surgical approach of the cervical spinal cord and brachial plexus was developed in nine dogs for avulsion and reimplantation of the ventral cervical spinal roots (C). The surgical steps involved in exposing the spinal cord and roots are described. The avulsed rootlets of C6 and C7 were reimplanted in their initial position. As a direct consequence of the avulsion, accid paralysis of the shoulder and severe amyotrophy developed within 5 ± 7 weeks on the injured side. In addition, the dogs exhibited clinical signs resulting from damage to long ®ber tracts due to the reimplantation procedure. A partial recovery of these de®cits was observed during the 6 postoperative months. Retrograde axonal tracing with horseradish peroxidase applied to the distal stump of the musculocutaneous, suprascapular, and subscapular nerves (originating from C5, C6 and C7) revealed the presence of labelled neuronal somata that were located in the ipsilateral ventral horn, close to the tip of the reimplanted rootlets. It is concluded that the dog constitutes a worthwhile animal model for the study of avulsion and reimplantation of brachial plexus root via a lateral surgical approach.

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“…Two advances to this technique expanded its utility. One was the use of tetramethylbenzidine (TMB) as a chromagen, which provided considerable amplification revealing both retrograde and anterograde transport of the tracer (Mesulam, 1978). The second advance was the conjugation of HRP with the plant lectin, wheat germ agglutinin (WGA-HRP) (Staines et al, 1980).…”

Section: Neuroanatomy; Axonal Tract Tracingmentioning

confidence: 99%

An anterograde neuroanatomical tracing method that shows the detailed morphology of neurons, their axons and terminals: Immunohistochemical localization of an axonally transported plant lectin,Phaseolus vulgaris leucoagglutinin (PHA-L)

1984

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A new neuroanatomical method for tracing connections in the central nervous system based on the anterograde axonal transport of the kidney bean lectin, Phaseolus vulgaris-leucoagglutinin (PHA-L) is described. The method, for which a detailed protocol is presented, offers several advantages over present techniques. First, when the lectin is delivered iontophoretically, PHA-L injection sites as small as 50-200 μm in diameter can be produced, and are clearly demarcated since the neurons within the labeled zone are completely filled. Second, many morphological features of such filled neurons are clearly demonstrated including their cell bodies, axons, dendritic arbors and even dendritic spines. Third, there is some evidence to suggest that only the neurons at the injection site that are filled transport demonstrable amounts of the tracer, raising the possibility that the effective injection site can be defined quite precisely. Fourth, even with the most restricted injections, the morphology of the labeled axons and axon terminals is clearly demonstrated; this includes boutons en passant, fine collateral branches, and various terminal specialization, all of which can be visualized as well as in the best rapid Golgi preparations. Fifth, when introduced iontophoretically, PHA-L appears to be transported preferentially in the anterograde direction; only rarely is it transported retrogradely. Sixth, PHA-L does not appear to be taken up and transported effectively by fibers of passage. Seventh, there is no discernible degradation of the transported PHA-L with survival times of up to 17 days. Finally, since the transported marker can be demonstrated with either peroxidase or fluorescent antibody techniques, it may be used in conjunction with other neuroanatomical methods. For example, double anterograde labeling experiments can be done using the autoradiographic method along with immunoperoxidase localization of PHA-L, and the retrogradely transported fluorescent dyes can be visualized in the same tissue sections as PHA-L localized with immunofluorescence techniques.

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Tetramethyl benzidine for horseradish peroxidase neurohistochemistry: a non-carcinogenic blue reaction product with superior sensitivity for visualizing neural afferents and efferents.

Cited by 4,268 publications

References 0 publications

Connections between the cochlear nuclei in guinea pig

Connections between the cochlear nuclei in guinea pig

Lateral approach of the dog brachial plexus for ventral root reimplantation

An anterograde neuroanatomical tracing method that shows the detailed morphology of neurons, their axons and terminals: Immunohistochemical localization of an axonally transported plant lectin,Phaseolus vulgaris leucoagglutinin (PHA-L)

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