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金鱼视觉系统中蛋白质p68/70的表达表明其在再生和神经发生中都发挥作用。

The expression of the protein p68/70 within the goldfish visual system suggests a role in both regeneration and neurogenesis.

作者信息

Wilmot G R, Raymond P A, Agranoff B W

机构信息

Neuroscience Program, University of Michigan, Ann Arbor 48104.

出版信息

J Neurosci. 1993 Jan;13(1):387-401. doi: 10.1523/JNEUROSCI.13-01-00387.1993.

Abstract

Previous attempts to characterize the molecular events that support successful regeneration of axotomized goldfish retinal ganglion cells (RGCs) have led to the discovery of an acidic protein doublet in retina that displays an increased incorporation of 35S-methionine following axotomy, and is known to be axonally transported within the optic nerve. This protein is termed p68/70, reflecting its relative migration in 10% SDS-PAGE. In the present study, an affinity-purified polyclonal antibody to p68/70 (anti-p68/70) was developed and used to assess the species, tissue, and cellular distribution of p68/70. The antibody cross-reacted with homogenates of brain and other tissues from goldfish and closely related fish species. While each goldfish tissue tested expressed p68/70, the levels varied over a 30-fold range, with the highest amounts in brain, egg, and ovary. Immunolabeling of goldfish retina revealed prominent staining of RGC somata, dendrites, and axons. During regeneration, the immunoreactivity of the RGC somata and axons increased dramatically. Intense immunolabeling was also observed in the germinal neuroepithelial cells and rod precursors and in all retinal layers near the peripheral margin, in the region of recently differentiated neurons. In the tectum, the germinal zone was also highly labeled. The elevated expression of p68/70 in each of these areas known to mediate neuronal growth within the goldfish visual system suggests that p68/70 plays a role in axonal growth, regrowth, and possibly in neural development as well.

摘要

此前,为了描述支持金鱼视网膜神经节细胞(RGCs)成功再生的分子事件,研究人员进行了多次尝试,结果发现视网膜中存在一种酸性蛋白双重体,在轴突切断后,该双重体对35S-甲硫氨酸的掺入增加,并且已知其在视神经中进行轴突运输。这种蛋白质被称为p68/70,反映了它在10%十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)中的相对迁移率。在本研究中,研究人员制备了一种针对p68/70的亲和纯化多克隆抗体(抗p68/70),并用于评估p68/70的物种、组织和细胞分布。该抗体与金鱼及亲缘关系密切的鱼类的脑和其他组织的匀浆发生交叉反应。虽然所检测的每个金鱼组织都表达p68/70,但其水平在30倍的范围内变化,在脑、卵和卵巢中的含量最高。金鱼视网膜的免疫标记显示,RGC的胞体、树突和轴突有明显的染色。在再生过程中,RGC胞体和轴突的免疫反应性显著增加。在生发神经上皮细胞、视杆前体细胞以及外周边缘附近所有视网膜层中,即在最近分化神经元的区域,也观察到强烈的免疫标记。在顶盖中,生发区也有高度标记。在金鱼视觉系统中,已知这些区域中的每一个都介导神经元生长,而p68/70在这些区域中的表达升高,这表明p68/70在轴突生长、再生以及可能在神经发育中发挥作用。

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