Transgenic mice expressing a dual, CRE-inducible reporter for the analysis of axon guidance and synaptogenesis
Transgenic mice expressing a dual, CRE-inducible reporter for the analysis of axon guidance and synaptogenesis
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DOI：
10.1002/dvg.20307
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发表时间：
2007-06-01
期刊：
影响因子：
1.5
通讯作者：
Giacomo Consalez, G.
中科院分区：
文献类型：
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作者：
Badaloni, Aurora;Bonanomi, Dario;Giacomo Consalez, G.
中枢神经系统轴突束的神经解剖剖析以及研究调控其组装和可塑性的细胞内在和细胞外在信号需要改进的模块化工具。在此我们描述了一种通用的转基因示踪剂，它可用于在胚胎神经管或出生后中枢神经系统的任何区域，以及任何野生型或突变型遗传背景下可视化轴突束和突触末端。该构建体允许CRE诱导表达一种双顺反子轴突标记物，其编码两种表面报告蛋白：一种法尼基化绿色荧光蛋白（farnesylated GFP）和人胎盘碱性磷酸酶（PLAP）。这两种蛋白都定位在神经元表面附近，从而能够同时检测同一神经元中的细胞体、神经突以及突触前和突触后位点。这为在体内对神经回路进行成像以及研究其在培养的神经元、神经干细胞和源自该报告基因系的组织外植体中的组装和重塑提供了一个CRE诱导的双重系统。与现有的品系不同，该报告基因不编码一个普遍表达的、可被loxP位点调控的LacZ基因，从而允许在突变品系中同时分析β -半乳糖苷酶的活性。
Improved and modular tools are needed for the neuroanatomical dissection of CNS axonal tracts, and to study the cell-intrinsic and cell-extrinsic cues that govern their assembly and plasticity. Here we describe a general purpose transgenic tracer that can be used to visualize axonal tracts and synaptic terminals in any region of the embryonic neural tube or postnatal CNS, on any wild type or mutant genetic background. The construct permits CRE-inducible expression of a dicistronic axonal marker encoding two surface reporter proteins: a farnesylated GFP and the human Placental Alkaline Phosphatase (PLAP). Both proteins localize alongside the neuronal surface, permitting the concomitant detection of cell body, neurites, and presynaptic and postsynaptic sites in the same neuron. This provides a CRE-inducible dual system for imaging neural circuits in vivo, and to study their assembly and remodeling in cultured neurons, neural stem cells, and tissue explants derived from the reporter line. Unlike existing lines, this reporter does not encode a ubiquitously expressed, floxable LacZ gene, permitting the simultaneous analysis of beta galactosidase activity in mutant lines.