Neural stem cells from mouse strain Thy1 YFP-16 are a valuable tool to monitor and evaluate neuronal differentiation and morphology
Neural stem cells from mouse strain Thy1 YFP-16 are a valuable tool to monitor and evaluate neuronal differentiation and morphology
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DOI：
10.1016/j.neulet.2016.10.001
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发表时间：
2016-11-10
影响因子：
2.5
通讯作者：
Mitrecic, Dinko
中科院分区：
文献类型：
--
作者：
Alic, Ivan;Kosi, Nina;Mitrecic, Dinko
为了分析脑中干细胞移植后的事件，需要用于追踪干细胞的强大工具。本研究利用136.Cg-Tg（Thy 1-YFP）16 Jrs/J（Thy 1 YFP-16）小鼠品系，其中黄色荧光蛋白（YFP）受Thy 1基因启动子控制。这允许可视化整个神经元，即它们的细胞体、轴突和树突。在这项工作中，荧光细胞在胚胎发育，体外分化，并在健康和中风影响的小鼠脑移植后。在胚胎发育过程中，Thy 1-YFP阳性细胞首先在E12.5出现，随后分布于前脑、菱脑、脊髓和周围神经。RT-PCR和免疫细胞化学定量分析显示，Thy 1-YFP阳性细胞在胚胎发育和体外分化过程中表达nestin和SOX 2，然后表达MAP 2、β 3-tubulin和NeuN。从E14.5分离的Thy 1-YFP阳性细胞占培养神经元的21.88 +/-053%（SD），并且这在体外分化过程中沿着保持恒定。另一方面，Thy 1-YFP阳性细胞的比例达到50%的神经元在围产期和一个月大的小鼠脑。通过YFP荧光可区分从Thy 1 YFP-16小鼠品系移植到健康和中风影响的脑的海马附近的神经干细胞。它们分化成成熟的神经元，甚至在移植后14周（我们实验的终点）也可以检测到。总之，来自Thy 1 YFP-16小鼠的干细胞是一种监测神经发生的出色工具，能够对新神经元及其投射进行形态学分析，特别是在脑中移植后。(C)2016作者由Elsevier爱尔兰有限公司发布。这是CC BY-NC-ND许可下的开放获取文章。
To analyse events following transplantation of stem cells in the brain robust tools for tracing stem cells are required. Here we took advantage of the mouse strain 136.Cg-Tg(Thy1-YFP)16Jrs/J (Thy1 YFP-16), where yellow fluorescent protein (YFP) is under control of the promoter of Thy1 gene. This allows visualising whole neurons, i.e. their cell body, axons and dendrites. In this work fluorescent cells were followed during embryonic development, in vitro differentiation, and after transplantation in the healthy and stroke-affected mouse brain. During embryonic development Thy1-YFP positive cells were first observed on E12.5 and subsequently located in the prosencephalon, rhombencephalon, spinal cord and peripheral nerves. Quantitative analysis by RT-PCR and immunocytochemistry revealed that Thy1-YFP positive cells during embryo development and in vitro differentiation were expressing nestin and SOX2 then MAP2, beta 3-tubulin and NeuN. Thy1-YFP positive cells isolated from E14.5 represented 21.88 +/- 053% (SD) of the cultivated neurons and this remained constant along in vitro differentiation. On the other hand, proportion of Thy1-YFP positive cells reached 50% of neurons in perinatal and one month old mouse brain. Neural stem cells isolated from Thy1 YFP-16 mouse strain transplanted near hippocampus of the healthy and stroke-affected brain were distinguishable by YFP fluorescence. They differentiated into mature neurons and were detectable even 14 weeks after transplantation, the end point of our experiment. In conclusion, stem cells originating from Thy1 YFP-16 mice represent an outstanding tool to monitor neurogenesis enabling morphological analyses of new neurons and their projections, in particular after transplantation in the brain. (C) 2016 The Authors. Published by Elsevier Ireland Ltd. This is an open access article under the CC BY-NC-ND license.