Retrograde labeling of retinal ganglion cells in adult zebrafish with fluorescent dyes.

Retrograde labeling of retinal ganglion cells in adult zebrafish with fluorescent dyes.
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用荧光染料逆行标记成年斑马鱼的视网膜神经节细胞。

DOI:
10.3791/50987
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发表时间:
2014-05
期刊:
Journal of Visualized Experiments
影响因子:
--
通讯作者:
Hu B.
Hu B.
中科院分区:
其他
文献类型:
--
作者:
Zou S.Q.;Tian C.;Du S.T.;Hu B.

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由于逆行标记视网膜神经节细胞(RGCs)可以将RGCs从死亡部位分离出来,已成为RGCs存活和再生实验中计数的金标准。许多研究已经在哺乳动物身上进行,以研究视神经损伤后视网膜神经节细胞的存活。然而,成年斑马鱼视网膜神经节细胞的逆行标记尚未见报道,尽管有一些替代方法可以计数视网膜神经节细胞层(RGCL)中的细胞数量。考虑到成年斑马鱼头骨体积小,在头骨上钻孔后死亡风险高,我们利用酸蚀的方法打开头骨,并用光固化粘合剂封孔,可以显著提高成活率。吸附5天后,几乎所有的RGC都被标记。由于该方法不需要横断视神经,在成年斑马鱼视神经损伤后视网膜节细胞存活的研究中具有不可替代的作用。在这里,我们逐步介绍这种方法,并给出有代表性的结果。
As retrograde labeling retinal ganglion cells (RGCs) can isolate RGCs somata from dying sites, it has become the gold standard for counting RGCs in RGCs survival and regeneration experiments. Many studies have been performed in mammalian animals to research RGCs survival after optic nerve injury. However, retrograde labeling of RGCs in adult zebrafish has not yet been reported, though some alternative methods can count cell numbers in retinal ganglion cell layers (RGCL). Considering the small size of the adult zebrafish skull and the high risk of death after drilling on the skull, we open the skull with the help of acid-etching and seal the hole with a light curing bond, which could significantly improve the survival rate. After absorbing the dyes for 5 days, almost all the RGCs are labeled. As this method does not need to transect the optic nerve, it is irreplaceable in the research of RGCs survival after optic nerve crush in adult zebrafish. Here, we introduce this method step by step and provide representative results.
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