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Using the Tg(nrd:egfp)/albino Zebrafish Line to Characterize In Vivo Expression of neurod

机译:使用Tg(nrd:egfp)/白化斑马鱼品系表征神经元的体内表达

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摘要

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.
机译: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,以进一步表征发育中和成年视网膜中神经元的表达,并确定成年感光细胞再生过程中神经元的表达。我们还提供了有关Neurod在多种其他组织中表达的观察结果。在该品系中,在发育中的和成年的视网膜,松果体,小脑,嗅球,中脑,后脑,神经管,侧线,内耳,胰腺,肠和鳍的细胞中发现了EGFP。使用免疫组织化学和原位杂交,我们比较了nrd:egfp转基因的表达与内源性神经元和已知的视网膜细胞类型的表达。与基于原位杂交的先前数据一致,我们显示了在视网膜发育过程中,nrd:egfp转基因在增殖性视网膜神经上皮细胞中不表达,而在一部分视网膜神经元中表达。与以前的研究相反,nrd:egfp在所有杆感光细胞中都逐渐表达。在成年斑马鱼的感光细胞再生过程中,原位杂交表明神经元在Müller胶质细胞来源的神经元祖细胞中不表达,但在它们迁移到外核层并分化为新的杆状感光细胞时在感光细胞祖细胞中表达。在光感受器再生过程中,nrd:egfp的表达与Neurod的表达匹配。我们得出的结论是,Tg(nrd:egfp)/白蛋白是内源性神经元表达的良好代表,是可视化各种组织中神经元表达的有用工具,并将有助于研究控制成人感光细胞再生的基本过程。

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