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棱镜 Plus:一种在四种不同脑细胞类型中表达不同荧光蛋白的小鼠品系。

PrismPlus: a mouse line expressing distinct fluorophores in four different brain cell types.

机构信息

Department of Neuroscience, University of Florida, Gainesville, FL, USA.

J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, FL, USA.

出版信息

Sci Rep. 2018 May 8;8(1):7182. doi: 10.1038/s41598-018-25208-y.

Abstract

To screen the complex central nervous system (CNS) injury responses, we created a quadruple-labelled 'PrismPlus' mouse line with a genetically encoded distinct fluorescent tag in oligodendrocytes, microglia, neurons, and astrocytes. Cx3cr1-gfp and Prism mice originally developed by Jung et al., 2000 and Dougherty et al., 2012, respectively, were cross-bred. First, we confirmed the presence of fluorophores in appropriate cell types in PrismPlus mice. PrismPlus mice were then used to examine the cellular responses to brain implanted micro-devices. We observed an increase in microglial response at earlier time points as compared to 4 weeks, a progressive astrocytic response, and fewer neurons at the vicinity of an implanted device. These results are similar to what has been described in literature using other rodent strains, previously attainable only through time-consuming and variable immunohistochemistry methods. Finally, we demonstrate the compatibility of PrismPlus brain tissue with CLARITY, an advanced tissue clearing technique, opening the door to future thick tissue imaging studies. This report confirms PrismPlus transgenic fluorescence and highlights the utility of these mice to study CNS injuries. The work herein seeks to establish a novel transgenic mouse line to improve experimental scope, consistency, and efficiency for CNS researchers.

摘要

为了筛选复杂的中枢神经系统(CNS)损伤反应,我们创建了一种四重标记的“PrismPlus”小鼠品系,其中寡突胶质细胞、小胶质细胞、神经元和星形胶质细胞中具有独特的遗传编码荧光标记。Cx3cr1-gfp 和 Prism 小鼠最初分别由 Jung 等人(2000 年)和 Dougherty 等人(2012 年)开发,通过杂交繁殖得到。首先,我们在 PrismPlus 小鼠中确认了适当细胞类型中荧光团的存在。然后,我们使用 PrismPlus 小鼠来检查大脑植入微设备后的细胞反应。与 4 周相比,我们观察到早期小胶质细胞反应增加,星形胶质细胞反应逐渐增加,植入设备附近的神经元减少。这些结果与使用其他啮齿动物品系在文献中描述的结果相似,以前只能通过耗时且可变的免疫组织化学方法获得。最后,我们证明了 PrismPlus 脑组织与 CLARITY(一种先进的组织透明技术)的兼容性,为未来的厚组织成像研究开辟了道路。本报告证实了 PrismPlus 转基因荧光,并强调了这些小鼠在研究 CNS 损伤方面的实用性。本文旨在建立一种新型的转基因小鼠品系,以提高 CNS 研究人员的实验范围、一致性和效率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/057c/5940666/c194883decee/41598_2018_25208_Fig1_HTML.jpg

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