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Immunocytochemical characterisation of ensheathing glia in the olfactory and vomeronasal systems of Ambystoma mexicanum (Caudata: Ambystomatidae)

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Abstract

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.

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References

  • Adams JC (1981) Heavy-metal intensification of DAB-based reaction product. J Histochem Cytochem 29:775

    Article  CAS  PubMed  Google Scholar 

  • Barnett SC, Riddell JS (2004) Olfactory ensheathing cells (OECs) and the treatment of CNS injury: advantages and possible caveats. J Anat 204:57–67

    Article  PubMed  PubMed Central  Google Scholar 

  • Barnett SC, Hutchins AM, Noble M (1993) Purification of olfactory nerve ensheathing cells from the olfactory bulb. Dev Biol 155:337–350

    Article  CAS  PubMed  Google Scholar 

  • Barraud P, Seferiadis AA, Tyson LD, Zwart MF, Szabo-Rogers HL, Ruhrberg C, Liu KJ, Baker CVH (2010) Neural crest origin of olfactory ensheathing glia. Proc Natl Acad Sci USA 107:21040–21045

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Berezin V (2009) Structure and function of the neural cell adhesion molecule NCAM. Advances in experimental medicine and biology, vol 663. Springer, New York

    Google Scholar 

  • Boruch AV, Conners JJ, Pipitone M, Deadwyler G, Storer PD, Devries GH, Jones KJ (2001) Neurotrophic and migratory properties of an olfactory ensheathing cell line. Glia 33:225–229

    Article  CAS  PubMed  Google Scholar 

  • Botero L, Gomez RM, Chaparro O (2013) Pathogenesis of spinal cord injuries and mechanisms of repair induced by olfactory ensheathing cells. Rev Neurol 56:521–531

    PubMed  Google Scholar 

  • Boyd JG, Doucette R, Kawaja MD (2005) Defining the role of olfactory ensheathing cells in facilitating axon remyelination following damage to the spinal cord. FASEB J 19:694–703

    Article  CAS  PubMed  Google Scholar 

  • Boyd JG, Jahed A, McDonald TG, Krol KM, Van Eyk JE, Doucette R, Kawaja MD (2006) Proteomic evaluation reveals that olfactory ensheathing cells but not Schwann cells express calponin. Glia 53:434–440

    Article  PubMed  Google Scholar 

  • Chamero P, Leinders-Zufall T, Zufall F (2012) From genes to social communication: molecular sensing by the vomeronasal organ. Trens Neurosci 35:597–606

    Article  CAS  Google Scholar 

  • Chuah MI, Au C (1991) Olfactory Schwann cells are derived from precursor cells in the olfactory epithelium. J Neurosci Res 29:72–180

    Article  Google Scholar 

  • Chung RS, Woodhouse A, Fung S, Dickson TC, West AH, Vickers JC, Chuah MI (2004) Olfactory ensheathing cells promote neurite sprouting of injured axons in vitro by direct cellular contact and secretion of soluble factors. Cell Mol Life Sci 61:1238–1245

    Article  CAS  PubMed  Google Scholar 

  • Coutts DJC, Humphries CE, Zhao C, Plant GW, Franklin RJM (2013) Embryonic-derived olfactory ensheathing cells remyelinate focal areas of spinal cord demyelination more efficiently than neonatal or adult-derived cells. Cell Transplant 22:1249–1261

    Article  PubMed  Google Scholar 

  • Cragnolini AB, Friedman WJ (2008) The function of p75NTR in glia. Trends Neurosci 31:99–104

    Article  CAS  PubMed  Google Scholar 

  • D’Aniello B, Pinelli C, Polese G, Luongo L, Rastogi RK (2008) Developmental analysis of the extrabulbar olfactory projections in the ranid frog with some phylogenetic considerations. Ital J Zool 75:21–28

    Article  Google Scholar 

  • De La Rosa-Prieto C, Saiz-Sanchez D, Ubeda-Bañon I, Argandoña-Palacios L, Garcia-Muñozguren S, Martinez-Marcos A (2009) Fate of marginal neuroblast in the vomeronasal epithelium of adult mice. J Comp Neurol 517:723–736

    Article  Google Scholar 

  • Dicke U, Roth G (2007) Evolution of the amphibian nervous system. In: Kaas JH (ed) Evolution of nervous systems, vol 2., Non-mammalian vertebratesAcademic Press, Oxford, pp 61–124

    Chapter  Google Scholar 

  • Eisthen LH, Polese G (2007) Evolution of vertebrate olfactory subsystems. In: Kaas JH (ed) Evolution of nervous systems, vol 2. Academic Press, Oxford, pp 355–406

    Chapter  Google Scholar 

  • Eisthen HL, Sengelaub DR, Schroeder DM, Alberts JR (1994) Anatomy and forebrain projections of the olfactory and vomeronasal organs in axolotls (Ambystoma mexicanum). Brain Behav Evol 44:108–124

    Article  CAS  PubMed  Google Scholar 

  • Fairless R, Barnett S (2005) Olfactory ensheathing cells: their role in central nervous system repair. Int J Bioch Cell Biol 37:693–699

    Article  CAS  Google Scholar 

  • Ferrando S, Gallus L, Gambardella C, Ghigliotti L, Ravera S, Vallarino M, Vacchi M, Tagliafierro G (2010a) Cell proliferation and apoptosis in the olfactory epithelium of the shark Scyliorhinus canicula. J Chem Neuroanat 40:293–300

    Article  CAS  PubMed  Google Scholar 

  • Ferrando S, Gallus L, Gambardella C, Vacchi M, Tagliafierro G (2010b) G protein alpha subunits in the olfactory epithelium of the holocephalan fish Chimaera monstrosa. Neurosci Lett 472:65–67

    Article  CAS  PubMed  Google Scholar 

  • Ferrando S, Gallus L, Gambardella C, Amaroli A, Cutolo A, Masini MA, Vallarino M, Vacchi M (2012) Neuronal nitric oxide synthase (nNOS) immunoreactivity in the olfactory system of a cartilaginous fish. J Chem Neuroanat 43:133–140

    Article  CAS  PubMed  Google Scholar 

  • Forni PE, Wray S (2012) Neural crest and olfactory system: new prospective. Mol Neurobiol 46:349–360

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Forni PE, Taylor-Burds C, Melvin VS, Williams T, Wray S (2011) Neural crest and ectodermal cells intermix in the nasal placode to give rise to GnRH-1 neurons, sensory neurons, and olfactory encheathing cells. J Neurosci 31:6915–6927

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Franceschini IA, Barnett SC (1996) Low-affinity NGF receptor and E-N-CAM expression define two types of olfactory nerve ensheathing cells that share a common lineage. Dev Biol 173:327–343

    Article  CAS  PubMed  Google Scholar 

  • Franklin RJ (2003) Remyelination by transplanted olfactory ensheathing cells. Anat Rec B New Anat 271:71–76

    Article  PubMed  Google Scholar 

  • Franssen EH, de Bree FM, Verhaagen J (2007) Olfactory ensheathing glia: their contribution to primary olfactory nervous system regeneration and their regenerative potential following transplantation into the injured spinal cord. Brain Res Rev 56:236–258

    Article  PubMed  Google Scholar 

  • Garbuglia M, Verzini M, Sorci G, Bianchi R, Giambanco I, Agneletti AL, Donato R (1999) The calcium-modulated proteins, S100A1 and S100B, as potential regulators of the dynamics of type III intermediate filaments. Braz J Med Biol Res 32:1177–1185

    Article  CAS  PubMed  Google Scholar 

  • Geller S, Kolasa E, Tillet Y, Duittoz A, Vaudin P (2013) Olfactory ensheathing cells form the microenvironment of migrating GnRH-1 neurons during mouse development. Glia 61:550–566

    Article  PubMed  Google Scholar 

  • Germanà A, Marino F, Guerrera MC, Campo S, De Girolamo P, Montalbano G, Germanà GP, Ochoa-Erena FJ, Ciriaco E, Vega JA (2008) Expression and distribution of S100 protein in the nervous system of the adult zebrafish (Danio rerio). Microsc Res Tech 71:248–255

    Article  PubMed  Google Scholar 

  • Granger N, Blamires H, Franklin RJM, Jeffery ND (2012) Autologous olfactory mucosal cell transplants in clinical spinal cord injury: a randomized double-blinded trial in a canine translational model. Brain 135:3227–3237

    Article  PubMed  PubMed Central  Google Scholar 

  • Graziadei PP, Graziadei GA (1979a) Neurogenesis and neuron regeneration in the olfactory system of mammals. I. Morphological aspects of differentiation and structural organization of the olfactory sensory neurons. J Neurocytol 8:1–18

    Article  CAS  PubMed  Google Scholar 

  • Graziadei GA, Graziadei PP (1979b) Neurogenesis and neuron regeneration in the olfactory system of mammals. II Degeneration and reconstitution of the olfactory neurons after axotomy. J Neurocytol 8:197–213

    Article  CAS  PubMed  Google Scholar 

  • Halpern M (2007) The evolution of the vomeronasal system. In: Kaas JH (ed) Evolution of nervous systems, vol 2., Non-mammalian vertebratesAcademic Press, Oxford, pp 407–415

    Chapter  Google Scholar 

  • Halpern M, Martinez-Marcos (2003) A Structure and function of the vomeronasal system: an update. Prog Neurobiol 70:245–318

    Article  CAS  PubMed  Google Scholar 

  • Hofmann MH, Meyer DL (1991) Functional subdivisions of the olfactory system correlate with lectin-binding properties in Xenopus. Brain Res 564:344–347

    Article  CAS  PubMed  Google Scholar 

  • Huang Q, Zhao S, Gaudin A, Quennedey B, Gascuel J (2005) Glial fibrillary acidic protein and vimentin expression in the frog olfactory system during metamorphosis. Neuro Rep 16:1439–1442

    CAS  Google Scholar 

  • Jahed A, Rowland JW, McDonald T, Boyd JG, Doucette R, Kawaja MD (2007) Olfactory ensheathing cells express smooth muscle alpha-actin in vitro and in vivo. J Comp Neurol 503:209–223

    Article  CAS  PubMed  Google Scholar 

  • Kalman M (1998) Asroglial architecture of the carp (Cyprinus carpio) brain as revealed by immunocytochemical staining against glial fibrillary acidic protein (GFAP). Anat Embryol 198:409–433

    Article  CAS  PubMed  Google Scholar 

  • Katoh H, Shibata S, Fukuda K, Sato M, Satoh E, Nagoshi N, Minematsu T, Matsuzaki Y, Akazawa C, Toyama Y, Nakamura M, Okano H (2011) The dual origin of the peripheral olfactory system: placode and neural crest. Mol Brain 4:34

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kawaja MD, Boyd JG, Smithson LJ, Jahed A, Doucette R (2009) Technical strategies to isolate olfactory ensheathing cells for intraspinal implantation. J Neurotrauma 26:155–177

    Article  PubMed  Google Scholar 

  • Key B, St John J (2002) Axon navigation in the mammalian primary olfactory pathway: where to next? Chem Senses 27:245–260

    Article  CAS  PubMed  Google Scholar 

  • Krudewig C, Deschl U, Wewetzer K (2006) Purification and in vitro characterization of adult canine olfactory ensheathing cells. Cell Tissue Res 326:687–696

    Article  CAS  PubMed  Google Scholar 

  • Lazzari M, Franceschini V (2005) Intermediate filament immunohistochemistry of astroglial cells in the leopard gecko, Eublepharis macularius. Anat Embryol 210:275–286

    Article  PubMed  Google Scholar 

  • Lazzari M, Franceschini V (2006) Glial cytoarchitecture in the central nervous system of the soft-shell turtle, Trionyx sinensis, revealed by intermediate filament immunohisto chemistry. Anat Embryol 211:497–506

    Article  PubMed  Google Scholar 

  • Lazzari M, Franceschini V, Ciani F (1997) Glial fibrillary acidic protein and vimentin in radial glia of Ambystoma mexicanum and Triturus carnifex: an immunocytochemical study. J Brain Res 38:187–194

    CAS  Google Scholar 

  • Lazzari M, Bettini S, Franceschini V (2013) Immunocytochemical characterization of olfactory ensheathing cells in fish. Brain Struct Funct 218:539–549

    Article  CAS  PubMed  Google Scholar 

  • Lazzari M, Bettini S, Franceschini V (2014) Immunocytochemical characterisation of olfactory ensheathing cells of zebrafish. J Anat 224:192–206

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li Y, Li D, Raisman G (2005a) Interaction of olfactory ensheathing cells with astrocytes may be the key to repair of tract injuries in the spinal cord: the ‘pathway hypothesis’. J Neurocytol 34:343–351

    Article  PubMed  Google Scholar 

  • Li Y, Field PM, Raisman G (2005b) Olfactory ensheathing cells and olfactory nerve fibroblasts maintain continuous open channels for regrowth of olfactory nerve fibres. Glia 52:245–251

    Article  PubMed  Google Scholar 

  • Lu J, Féron F, Mackay-Sim A, Waite PM (2002) Olfactory ensheathing cells promote locomotor recovery after delayed transplantation into transacted spinal cord. Brain 125:14–21

    Article  PubMed  Google Scholar 

  • Lucero MT (2013) Peripheral modulation of smell: fact or fiction? Semin Cell Dev Biol 24:58–70

    Article  PubMed  PubMed Central  Google Scholar 

  • Mackay-Sim A (2005) Olfactory ensheathing cells and spinal cord repair. Keio J Med 54:8–14

    Article  PubMed  Google Scholar 

  • Mackay-Sim A, Kittel PW (1991) On the life span of olfactory receptor neurons. Eur J Neurosci 3:209–215

    Article  PubMed  Google Scholar 

  • Maselli V, Polese G, Larson G, Raia P, Forte N, Rippa D, Ligrone R, Vicidomini R, Fulgione D (2014) A dysfunctional sense of smell: the irreversibility of olfactory evolution in free-living pigs. Evol Biol 41:229–239

    Article  Google Scholar 

  • Mayeur A, Duclos C, Honoré A, Gauberti M, Drouot L, do Rego JC, Bon-Mardion N, Jean L, Vérin E, Emery E, Lemarchant S, Vivien D, Boyer Q, Marie JP, Guérout N (2013) Potential of olfactory ensheathing cells from different sources for spinal cord repair. PLoS One 8:e62860

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Miragall F, Kadmon G, Schachner M (1989) Expression of L1 and N-CAM cell adhesion molecules during development of the mouse olfactory system. Dev Biol 135:272–286

    Article  CAS  PubMed  Google Scholar 

  • Nakajima M, Tsuruta M, Mori H, Nishikawa C, Okuyama S, Furukawa Y (2013) A comparative study of axon-surrounding cells in the two nasal nerve tracts from mouse olfactory epithelium and vomeronasal organ. Brain Res 1503:16–23

    Article  CAS  PubMed  Google Scholar 

  • Panni P, Ferguson IA, Beacham I, Mackay-Sim A, Ekberg JAK, St John JA (2013) Phagocytosis of bacteria by olfactory ensheathing cells and Schwann cells. Neurosci Lett 539:65–70

    Article  CAS  PubMed  Google Scholar 

  • Pellitteri R, Spatuzza M, Stanzani S, Zaccheo D (2010) Biomarkers expression in rat olfactory ensheathing cells. Front Biosci S2:289–298

    Article  CAS  Google Scholar 

  • Pinelli C, D’Aniello B, Polese G, Rastogi RK (2004) Extrabulbar olfactory system and nervus terminalis FMRFamide immunoreactive components in Xenopus laevis ontogenesis. J Chem Neuroanat 28:37–46

    Article  CAS  PubMed  Google Scholar 

  • Pixley SK (1996) Characterization of olfactory receptor neurons and other cell types in dissociated rat olfactory cell cultures. Int J Dev Neurosci 14:823–839

    Article  CAS  PubMed  Google Scholar 

  • Puche AC, Key B (1996) N-acetyl-lactosamine in the rat olfactory system: expression and potential role in neurite growth. J Comp Neurol 364:267–278

    Article  CAS  PubMed  Google Scholar 

  • Puche AC, Poirier F, Hair M, Berlett PF, Key B (1996) Role of galectin-1 in the developing mouse olfactory system. Dev Biol 179:274–287

    Article  CAS  PubMed  Google Scholar 

  • Quintana-Urzainqui I, Rodrıguez-Moldes I, Candal E (2014) Developmental, tract-tracing and immunohistochemical study of the peripheral olfactory system in a basal vertebrate: insights on Pax6 neurons migrating along the olfactory nerve. Brain Struct Funct 219:85–104

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Radtke C, Akiyama Y, Brokaw J, Lankford KL, Wewetzer K, Fodor WL, Kocsis JD (2004) Remyelination of the nonhuman primate spinal cord by transplantation of H-transferase transgenic adult pig olfactory ensheathing cells. FASEB J 18:335–337

    CAS  PubMed  PubMed Central  Google Scholar 

  • Raisman G (2001) Olfactory ensheathing cells: another miracle cure for spinal cord injury. Nat Rev Neurosci 2:369–375

    Article  CAS  PubMed  Google Scholar 

  • Raisman G (2007) Repair of spinal cord injury by transplantation of olfactory ensheathing cells. C R Biol 330:557–560

    Article  PubMed  Google Scholar 

  • Ramon-Cueto A, Avila J (1998) Olfactory ensheathing cells: properties and function. Brain Res Bull 46:175–187

    Article  CAS  PubMed  Google Scholar 

  • Rawji KS, Zhang SX, Tsai Y-Y, Smithson LJ, Kawaja MD (2013) Olfactory ensheathing cells of hamsters, rabbits, monkeys, and mice express α-smooth muscle actin. Brain Res 1521:31–50

    Article  CAS  PubMed  Google Scholar 

  • Roth FC, Laberge F (2011) High convergence of olfactory and vomeronasal influence in the telencephalon of the terrestrial salamander Plethodon shermani. Neuroscience 177:148–158

    Article  CAS  PubMed  Google Scholar 

  • Santamaria-Kisiel L, Rintala-Dempsey AC, Shaw GS (2006) Calcium-dependent and -independent interactions of the S100 protein family. Biochem J 396:201–214

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Smithson LJ, Kawaja MD (2009) A comparative examination of biomarkers for olfactory ensheathing cells in cats and guinea pigs. Brain Res 1284:41–53

    Article  CAS  PubMed  Google Scholar 

  • St John JA, Key B (1999) Expression of galectin-1 in the olfactory nerve pathway of rat. Dev Brain Res 117:171–178

    Article  CAS  Google Scholar 

  • Su Z, Chen J, Qiu Y, Yuan Y, Zhu F, Zhu Y, Liu X, Pu Y, He C (2013) Olfactory ensheathing cells: the primary innate immunocytes in the olfactory pathway to engulf apoptotic olfactory nerve debris. Glia 61:490–503

    Article  PubMed  Google Scholar 

  • Suzuki J, Yoshizaki K, Kobayashi T, Osumi N (2013) Neural crest-derived horizontal basal cells as tissue stem cells in the adult olfactory epithelium. Neurosci Res 75:112–120

    Article  PubMed  Google Scholar 

  • Techangamsuwan S, Kreutzer R, Kreutzer M, Imbschweiler I, Rohn K, Wewetzer K, Baumgartner W (2009) Transfection of adult canine Schwann cells and olfactory ensheathing cells at early and late passage with human TERT differentially affects growth factor responsiveness and in vitro growth. J Neurosci Methods 176:112–120

    Article  CAS  PubMed  Google Scholar 

  • Vincent AJ, West AK, Chuah MI (2005) Morphological and functional plasticity of olfactory ensheathing cells. J Neurocytol 34:65–80

    Article  PubMed  Google Scholar 

  • von Bartheld CS (2004) The terminal nerve and its relation with extrabulbar “olfactory” projections: lessons from lampreys and lungfishes. Microsc Res Tech 65:13–24

    Article  Google Scholar 

  • Woodhall E, West AK, Chuah MI (2001) Cultured olfactory ensheathing cells express nerve growth factor, brain-derived neurotrophic factor, glia cell line-derived neurotrophic factor and their receptors. Mol Brain Res 88:203–213

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study was supported by grants from the Italian Ministry of University and Research: FFO2009, FFO2010.

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  1. Department of Biological Geological and Environmental Sciences, University of Bologna, Via F. Selmi 3, 40126, Bologna, Italy

    Maurizio Lazzari, Simone Bettini & Valeria Franceschini

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  1. Maurizio Lazzari
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  2. Simone Bettini
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Correspondence to Maurizio Lazzari.

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Lazzari, M., Bettini, S. & Franceschini, V. Immunocytochemical characterisation of ensheathing glia in the olfactory and vomeronasal systems of Ambystoma mexicanum (Caudata: Ambystomatidae). Brain Struct Funct 221, 955–967 (2016). https://doi.org/10.1007/s00429-014-0949-8

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