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The knowledge of the morphological and functional aspects of mammalian glial cells has greatly increased in the last few decades. Glial cells represent the most diffused cell type in the central nervous system, and they play a critical role in the development and function of the brain. Glial cell dysfunction has recently been shown to contribute to various neurological disorders, such as autism, schizophrenia, pain, and neurodegeneration. For this reason, glia constitutes an interesting area of research because of its clinical, diagnostic, and pharmacological relapses. In this chapter, we present and discuss the cytoarchitecture of glial cells in tetrapods from an evolutive perspective. GFAP and vimentin are main components of the intermediate filaments of glial cells and are used as cytoskeletal molecular markers because of their high degree of conservation in the various vertebrate groups. In the anamniotic tetrapods and their progenitors, Rhipidistia (Dipnoi are the only extant rhipidistian fish), the cytoskeletal markers show a model based exclusively on radial glial cells. In the transition from primitive vertebrates to successively evolved forms, the emergence of a new model has been observed which is believed to support the most complex functional aspects of the nervous system in the vertebrates. In reptiles, radial glial cells are prevalent, but star-shaped astrocytes begin to appear in the midbrain. In endothermic amniotes (birds and mammals), star-shaped astrocytes are predominant. In glial cells, vimentin is indicative of immature cells, while GFAP indicates mature ones.Olfactory receptor neurons undergo continuous turnover, so they are an easy model for neurogenesis studies. Moreover, they are useful in neurotoxicity studies because of the exposed position of their apical pole to the external environment. Among vertebrates, fish represent a valid biological model in this field. In particular, zebrafish, already used in laboratories for embryological, neurobiological, genetic, and pathophysiological studies, is the reference organism in olfactory system research. Smell plays an important role in the reproductive behavior of fish, with direct influences also on the numerical consistency of their populations. Taking into account that a lot of species have considerable economic importance, it is necessary to verify if the model of zebrafish olfactory organ is also directly applicable to other fish. In this chapter, we focus on crypt cells, a morphological type of olfactory cells specific of fish. We describe hypothetical function (probably related with social behavior) and evolutive position of these cells (prior to the appearance of the vomeronasal organ in tetrapods). We also offer the first comparison of the molecular characteristics of these receptors between zebrafish and the guppy. Interestingly, the immunohistochemical expression patterns of known crypt cell markers are not overlapping in the two species.

Olfactory crypt neurons have been observed in several bony fishes and chondrichtyans. Although their morphology is uniform in all fish, very few is known about their antigenic properties, usually studied in zebrafish, but quite overlooked in other species. We tested in Poecilia reticulata (guppy) the two antibodies recognized to mark zebrafish crypt cells: while anti-S100 showed an immunohistochemical pattern comparable to what reported in zebrafish, anti-TrkA gave no signal. Western blot analysis revealed that S100-antiserum bound an antigen of expected weight, probably belonging to the S100 family. On the contrary, anti-TrkA detected more bands, but the protein/s might be too much diffused and/or diluted in the tissue to be detected with immunohistochemistry. Because of the high level of conservation in the Trk family proteins of the kinase domain, on which anti-TrkA was produced, we also tested anti-TrkB to exclude cross reactivity. Immunohistochemistry and Western blot confirmed that anti-TrkB displayed high specificity to its target and a different staining pattern compared to anti-TrkA, but, as anti-TrkA, it did not label crypt neurons. Finally, we documented that calretinin, a known marker of zebrafish ciliated and microvillous olfactory cells, in the guppy is expressed also by a subpopulation of S100-positive crypt neurons. These results reveal differences in antigen expression between zebrafish and guppy crypt cells. Together with the already known species-specific projections to the olfactory bulb and a heterogeneous panel of odorants, our findings support the possibility that crypt cells are functionally less uniform as supposed.

The olfactory and vomeronasal systems of vertebrates are characterised by neurogenesis occurring throughout life. The regenerative ability of olfactory receptor neurons relies on specific glial cells, the olfactory and vomeronasal axon-surrounding cells. Numerous studies have examined mammalian olfactory ensheathing cells which are considered potential candidates for spinal cord injury repair using cell-based therapy. With regard to non-mammalian vertebrates, limited information is available on these glial cells in fish, and there is no information on them in terrestrial anamniotes, the amphibians. In the present research, we studied the immunocytochemical characteristics of axon-surrounding cells in Ambystoma mexicanum. Urodeles have relatively simple olfactory and vomeronasal systems, and represent a good model for studying ensheathing cells in extant representatives of basal tetrapods. Sections from the decalcified heads of A. mexicanum were immunocytochemically processed for the detection of proteins used in research on mammalian olfactory-ensheathing cells. S100, GFAP and NCAM were clearly observed. p75NTR, Gal-1 and PSA-NCAM showed weak staining. No vimentin immunopositivity was observed. The corresponding areas of the olfactory and vomeronasal pathways displayed the same staining characteristics, with the exception of Gal-1, p75NTR and PSA-NCAM in the mucosae. The degree of marker expression was not uniform throughout the sensory pathways. In contrast to fish, both olfactory and vomeronasal nerves displayed uniform staining intensity. This study showed that some markers for mammalian and fish-ensheathing glia are also applicable in urodeles. The olfactory systems of vertebrates show similarities, and also clear dissimilarities. Further investigations are required to ascertain the functional significance of these regional and interspecific differences.