Laboratory of Molecular Neurosurgery, Department of Stereotactic Neurosurgery, University Hospital Freiburg, Breisacher Str. 64, 79106 Freiburg, Germany.
J Neurosci Methods. 2011 Apr 15;197(1):6-13. doi: 10.1016/j.jneumeth.2011.01.015. Epub 2011 Feb 3.
Huntington's disease (HD) is a neurodegenerative disease with currently only symptomatic treatment. Cell-based therapy, aiming at replacing the lost medium spiny neurons (MSN) with primary fetal striatal cells, has had some success at modifying the symptoms both in experimental studies and clinical trials. Additional pre-clinical studies are required to optimise transplantation protocols and conditions, learn about the limits of circuit reconstruction and functional recovery, and test alternative cell sources. Transgenic mice with integrated bacterial artificial chromosome (BAC) expressing the green fluorescent protein (GFP) can be used to study specific neuronal projections. The BAC transgenic line used in this study, with the GFP expression under the control of the muscarinic receptor M4 promoter, selectively expressed the reporter gene in the direct efferent pathway of the MSN projecting from the striatum to the substantia nigra pars reticulata and the entopeduncular nucleus, the rodent equivalent of the internal segment of the globus pallidus. The current work was designed to validate the use of M4-BAC-GFP mice as tissue donors in cell-based therapy in a rodent model of HD by examining the effect of the transplantation procedure on the GFP expression; the feasibility of identifying the GFP expression in vivo after different time points; and the survival and integration of the transgenic striatal tissue transplant up to 6 months in the host. The data confirm that embryonic striatal tissue from the M4-BAC-GFP mice survives, stably expresses GFP, and thus represents a powerful novel way to study graft-host interaction in this animal model neurodegeneration.
亨廷顿病（HD）是一种神经退行性疾病，目前仅对症治疗。基于细胞的疗法旨在用原代胎鼠纹状体细胞替代丢失的中型棘神经元（MSN），在实验研究和临床试验中都在改善症状方面取得了一定的成功。需要进行更多的临床前研究来优化移植方案和条件，了解回路重建和功能恢复的限制，并测试替代细胞来源。整合表达绿色荧光蛋白（GFP）的细菌人工染色体（BAC）的转基因小鼠可用于研究特定的神经元投射。本研究中使用的 BAC 转基因系，GFP 的表达受毒蕈碱受体 M4 启动子的控制，在从中脑纹状体投射到黑质网状部和被盖脚核的 MSN 的直接传出通路上选择性地表达报告基因，该通路由纹状体投射到黑质网状部和被盖脚核，是啮齿动物相当于苍白球内节的投射。目前的工作旨在通过检查移植程序对 GFP 表达的影响，鉴定 M4-BAC-GFP 小鼠作为细胞治疗中 HD 啮齿动物模型组织供体的用途，来验证该方法的可行性；以及鉴定 GFP 表达在不同时间点的体内可行性；以及在宿主中长达 6 个月的转基因纹状体组织移植的存活和整合。数据证实，来自 M4-BAC-GFP 小鼠的胚胎纹状体组织存活，稳定表达 GFP，因此代表了研究该动物模型神经退行性变中移植物-宿主相互作用的一种强有力的新方法。
