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斑马鱼嗅鞘细胞的免疫细胞化学特征分析

Immunocytochemical characterisation of olfactory ensheathing cells of zebrafish.

作者信息

Lazzari Maurizio, Bettini Simone, Franceschini Valeria

机构信息

Department of Biological, Geological and Environmental Sciences, University of Bologna, Bologna, Italy.

出版信息

J Anat. 2014 Feb;224(2):192-206. doi: 10.1111/joa.12129. Epub 2013 Oct 24.

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.

摘要

持续终生的神经发生是脊椎动物嗅觉系统的典型特征。嗅觉受体神经元的再生能力依赖于嗅觉通路特有的神经胶质细胞类型,即“嗅觉鞘细胞”。由于其在基于细胞的脊髓损伤修复治疗中的潜在作用,迄今为止已有多项研究聚焦于哺乳动物的嗅觉鞘细胞。然而,关于非哺乳动物脊椎动物,尤其是无羊膜动物中这种神经胶质细胞类型的信息有限。在当前的免疫细胞化学研究中,我们分析了斑马鱼(Danio rerio)嗅觉鞘细胞的特征。鱼类为研究与非哺乳动物脊椎动物嗅觉通路相关的神经胶质细胞提供了一个良好的模型。特别是,斑马鱼具有许多有价值的特征,使其能够作为遗传、神经生物学和发育研究以及毒理学和基因组学研究的主要模式生物。对斑马鱼脱钙头部的石蜡切片进行免疫细胞化学处理,以检测用于哺乳动物嗅觉鞘细胞研究的蛋白质,包括胶质纤维酸性蛋白(GFAP)、S100、神经细胞黏附分子(NCAM)、多唾液酸NCAM(PSA-NCAM)、波形蛋白(VIM)、p75神经营养因子受体(p75NTR)和半乳糖凝集素(Gal)-1。值得注意的是,清晰观察到了GFAP、S100、NCAM和Gal-1,而未检测到波形蛋白染色。PSA-NCAM和p75NTR的免疫染色较弱。此外,斑马鱼嗅觉通路各区域的标记物表达程度并不一致。斑马鱼嗅觉系统的免疫染色模式在一定程度上与其他鱼类不同,表明存在种间差异。我们还表明,斑马鱼的嗅觉通路表达哺乳动物嗅觉鞘细胞的标记物。脊椎动物的嗅觉系统有相似之处,但也存在显著差异。嗅觉通路标记物免疫染色模式的区域和种间差异是否具有功能意义这一问题需要进一步研究。

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