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利用 Tg(nrd:egfp)/albino 斑马鱼系来描述神经调节蛋白在体内的表达。

Using the Tg(nrd:egfp)/albino zebrafish line to characterize in vivo expression of neurod.

机构信息

Department of Anatomy and Cell Biology and Department of Ophthalmology, Wayne State University School of Medicine, Detroit, Michigan, United States of America.

出版信息

PLoS One. 2012;7(1):e29128. doi: 10.1371/journal.pone.0029128. Epub 2012 Jan 3.

Abstract

In this study, we used a newly-created transgenic zebrafish, Tg(nrd:egfp)/albino, to further characterize the expression of neurod in the developing and adult retina and to determine neurod expression during adult photoreceptor regeneration. We also provide observations regarding the expression of neurod in a variety of other tissues. In this line, EGFP is found in cells of the developing and adult retina, pineal gland, cerebellum, olfactory bulbs, midbrain, hindbrain, neural tube, lateral line, inner ear, pancreas, gut, and fin. Using immunohistochemistry and in situ hybridization, we compare the expression of the nrd:egfp transgene to that of endogenous neurod and to known retinal cell types. Consistent with previous data based on in situ hybridizations, we show that during retinal development, the nrd:egfp transgene is not expressed in proliferating retinal neuroepithelium, and is expressed in a subset of retinal neurons. In contrast to previous studies, nrd:egfp is gradually re-expressed in all rod photoreceptors. During photoreceptor regeneration in adult zebrafish, in situ hybridization reveals that neurod is not expressed in Müller glial-derived neuronal progenitors, but is expressed in photoreceptor progenitors as they migrate to the outer nuclear layer and differentiate into new rod photoreceptors. During photoreceptor regeneration, expression of the nrd:egfp matches that of neurod. We conclude that Tg(nrd:egfp)/albino is a good representation of endogenous neurod expression, is a useful tool to visualize neurod expression in a variety of tissues and will aid investigating the fundamental processes that govern photoreceptor regeneration in adults.

摘要

在这项研究中,我们使用了一种新创建的转基因斑马鱼,Tg(nrd:egfp)/albino,进一步研究了 neurod 在发育中和成年视网膜中的表达,并确定了 neurod 在成年光感受器再生过程中的表达。我们还提供了关于 neurod 在多种其他组织中表达的观察结果。在这条线中,EGFP 存在于发育中和成年视网膜、松果体、小脑、嗅球、中脑、后脑、神经管、侧线、内耳、胰腺、肠道和鳍的细胞中。通过免疫组织化学和原位杂交,我们将 nrd:egfp 转基因的表达与内源性 neurod 的表达以及已知的视网膜细胞类型进行了比较。与基于原位杂交的先前数据一致,我们表明在视网膜发育过程中,nrd:egfp 转基因不在增殖的视网膜神经上皮中表达,而是在一组视网膜神经元中表达。与先前的研究不同,nrd:egfp 在所有杆状光感受器中逐渐重新表达。在成年斑马鱼的光感受器再生过程中,原位杂交揭示了 neurod 不在 Muller 胶质衍生的神经元祖细胞中表达,但在光感受器祖细胞中表达,因为它们迁移到外核层并分化为新的杆状光感受器。在光感受器再生过程中,nrd:egfp 的表达与 neurod 的表达相匹配。我们得出结论,Tg(nrd:egfp)/albino 很好地代表了内源性 neurod 的表达,是一种可视化各种组织中 neurod 表达的有用工具,并将有助于研究控制成年光感受器再生的基本过程。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6200/3250405/1a4136990c3f/pone.0029128.g001.jpg

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