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  • Immunocytochemical characterisation of olfactory ensheathing cells of zebrafish.

Immunocytochemical characterisation of olfactory ensheathing cells of zebrafish.

Journal of anatomy (2013-10-30)
Maurizio Lazzari, Simone Bettini, Valeria Franceschini
ABSTRACT

Continuous lifelong neurogenesis is typical of the vertebrate olfactory system. The regenerative ability of olfactory receptor neurons is dependent on the glial cell type specific to the olfactory pathway, designated 'olfactory ensheathing cells'. Several studies to date have focused on mammalian olfactory ensheathing cells, owing to their potential roles in cell-based therapy for spinal cord injury repair. However, limited information is available regarding this glial cell type in non-mammalian vertebrates, particularly anamniotes. In the current immunocytochemical study, we analysed the features of olfactory ensheathing cells in the zebrafish, Danio rerio. Fish provide a good model for studying glial cells associated with the olfactory pathway of non-mammalian vertebrates. In particular, zebrafish has numerous valuable features that enable its use as a prime model organism for genetic, neurobiological and developmental studies, as well as toxicology and genomics research. Paraffin sections from decalcified heads of zebrafish were processed immunocytochemically to detect proteins used in the research on mammalian olfactory ensheathing cells, including glial fibrillary acid protein (GFAP), S100, neural cell adhesion molecule (NCAM), polysialylated NCAM (PSA-NCAM), vimentin (VIM), p75NTR and galactin (Gal)-1. Notably, GFAP, S100, NCAM and Gal-1 were clearly observed, whereas no vimentin staining was detected. Weak immunostaining for PSA-NCAM and p75NTR was evident. Moreover the degree of marker expression was not uniform in various tracts of the zebrafish olfactory pathway. The immunostaining patterns of the zebrafish olfactory system are distinct from those of other fish to some extent, suggesting interspecific differences. We also showed that the olfactory pathway of zebrafish expresses markers of mammalian olfactory ensheathing cells. The olfactory systems of vertebrates have similarities but there are also marked variations between them. The issue of whether regional and interspecific differences in immunostaining patterns of olfactory pathway markers have functional significance requires further investigation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Goat Anti-Mouse IgMµ Antibody, HRP conjugate, Upstate®, from goat
Sigma-Aldrich
Anti-p75NTR (Neurotrophin Receptor) Antibody, Upstate®, from rabbit
Sigma-Aldrich
Anti-LGALS1 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
Sigma-Aldrich
Anti-Mouse IgG (whole molecule)–Peroxidase antibody produced in goat, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Anti-Polysialic Acid-NCAM Antibody, clone 2-2B, ascites fluid, clone 2-2B, Chemicon®
Sigma-Aldrich
Anti-Neural Cell Adhesion Molecule Antibody, Chemicon®, from rabbit
Sigma-Aldrich
Anti-Vimentin Antibody, clone V9, clone V9, Chemicon®, from mouse