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.2009 Mar 30;178(1):1-9.
doi: 10.1016/j.jneumeth.2008.11.003. Epub 2008 Nov 13.

Versatile, high-resolution anterograde labeling of vagal efferent projections with dextran amines

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Versatile, high-resolution anterograde labeling of vagal efferent projections with dextran amines

Gary C Walter et al. J Neurosci Methods..

Abstract

None of the anterograde tracers used to label and investigate vagal preganglionic neurons projecting to the viscera has proved optimal for routine and extensive labeling of autonomic terminal fields. To identify an alternative tracer protocol, the present experiment evaluated whether dextran conjugates, which have produced superior results in the CNS, might yield widespread and effective labeling of long, fine-caliber vagal efferents in the peripheral nervous system. The dextran conjugates that were evaluated proved reliable and versatile for labeling the motor neuron pool in its entirety, for single- and multiple-labeling protocols, for both conventional and confocal fluorescence microscopy, and for permanent labeling protocols for brightfield microscopy of the projections to the gastrointestinal (GI) tract. Using a standard ABC kit followed by visualization with DAB as the chromagen, Golgi-like labeling of the vagal efferent terminal fields in the GI wall was achieved with the biotinylated dextrans. The definition of individual terminal varicosities was so sharp and detailed that it was routinely practical to examine the relationship of putative vagal efferent contacts (by the criteria of high magnification light microscopy) with the dendritic and somatic architecture of counterstained neurons in the myenteric plexus. Overall, dextran conjugates provide high-definition labeling of an extensive vagal motor pool in the GI tract, and offer considerable versatility when multiple-staining protocols are needed to elucidate the complexities of the innervation of the gut.

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Figures

Figure 1
Figure 1
A photomontage illustrating dense vagal efferent innervation of the myenteric plexus in the smooth muscle wall of the stomach following injection of dextran-tetramethylrhodamine-biotin into the dorsal motor nucleus of the vagus (dmnX). The fluorescent labeling was photographed and then image was inverted to better illustrate the morphology of individual fibers. Scale bar = 100 μm.
Figure 2
Figure 2
Counterstaining of the myenteric plexus to label either the total population of neurons (A,B) or the nitrergic subpopulation of neurons (C–D) provided excellent contrast to efferent axons and terminals labeled with the anterograde tracer dextran-biotin visualized using a routine DAB protocol. Fibers of passage, distinguishable by their smooth appearance, were routinely observed passing through and within close proximity to ganglia (A,C,F). Vagal efferents formed distinct “basket-like” endings around both nitrergic (C,E) and presumed cholinergic (F) neurons. Dextrans were just as effective in aging rats (G) as they were in younger rats. Scale bar = 16 μm in F for A, B; 25 μm for D-F; Scale bars in C and G = 16 μm.
Figure 3
Figure 3
A: Double labeling of vagal efferent (green) and vagal afferent (red) innervation of myenteric ganglia was readily achieved by injecting dextran-biotin into the dmnx and dextran-Texas Red into the nodose ganglia. Whole mounts were further processed in the presence of streptavidin Alexa Fluor 488.B: Triple labeling was accomplished using dextran-Texas Red tracing of vagal efferents (red) combined with alpha-synuclein (green) and NOS (blue) immunohistochemical labeling of myenteric neurons in the stomach. Prominent yellow varicosities indicate co-localization of alpha-synuclein with dextran-Texas Red in putative vagal efferent terminals. Scale bars for A = 30 μm; Scale bars for B = 50 μm.
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References

    1. Berthoud HR. Anatomical demonstration of vagal input to nicotinamide acetamide dinucleotide phosphate diaphorase-positive (nitrergic) neurons in rat fundic stomach. J Comp Neurol. 1995;358:428–439. - PubMed
    1. Berthoud HR. Morphological analysis of vagal input to gastrin releasing peptide and vasoactive intestinal peptide containing neurons in the rat glandular stomach. J Comp Neurol. 1996;370:61–70. - PubMed
    1. Berthoud HR, Carlson NR, Powley TL. Topography of efferent vagal innervation of the rat gastrointestinal tract. Am J Physiol. 1991;260:R200–207. - PubMed
    1. Berthoud HR, Jedrzejewska A, Powley TL. Simultaneous labeling of vagal innervation of the gut and afferent projections from the visceral forebrain with dil injected into the dorsal vagal complex in the rat. J Comp Neurol. 1990;301:65–79. - PubMed
    1. Chang HT. Immunoperoxidase labeling of the anterograde tracer fluoro-ruby (tetramethylrhodamine-dextran amine conjugate) Brain Res Bull. 1993;30:115–118. - PubMed

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