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Monoclonal L‐Citrulline Immunostaining Reveals Nitric Oxide‐Producing Vestibular Neurons
Author(s) -
HOLSTEIN GAY R.,
FRIEDRICH VICTOR L.,
MARTINELLI GIORGIO P.
Publication year - 2001
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.2001.tb03736.x
Subject(s) - vestibular nuclei , medial vestibular nucleus , nitric oxide , citrulline , nitric oxide synthase , immunocytochemistry , lateral vestibular nucleus , immunostaining , chemistry , biology , vestibular system , microbiology and biotechnology , neuroscience , anatomy , immunohistochemistry , biochemistry , endocrinology , immunology , arginine , amino acid
A bstract : Nitric oxide is an unstable free radical that serves as a novel messenger molecule in the central nervous system (CNS). In order to understand the interplay between classic and novel chemical communication systems in vestibular pathways, the staining obtained using a monoclonal antibody directed against L‐citrulline was compared with the labeling observed using more traditional markers for the presence of nitric oxide. Brainstem tissue from adult rats was processed for immunocytochemistry employing a monoclonal antibody directed against L‐citrulline, a polyclonal antiserum against neuronal nitric oxide synthase, and/or NADPH‐diaphorase histochemistry. Our findings demonstrate that L‐citrulline can be fixed in situ by vascular perfusion, and can be visualized in fixed CNS tissue sections by immunocytochemistry. Further, the same vestibular regions and cell types are labeled by NADPH‐diaphorase histochemistry, by the neuronal nitric oxide synthase antiserum, and by our anti‐L‐citrulline antibody. Clusters of L‐citrulline‐immunoreactive neurons are present in subregions of the vestibular nuclei, including the caudal portion of the inferior vestibular nucleus, the magnocellular portion of the medial vestibular nucleus, and the large cells in the ventral tier of the lateral vestibular nucleus. NADPH‐diaphorase histochemical staining of these neurons clearly demonstrated their multipolar, fusiform and globular somata and long varicose dendritic processes. These results provide support for the suggestion that nitric oxide serves key roles in both vestibulo‐autonomic and vestibulo‐spinal pathways.