Transneuronal tracing of diverse CNS circuits by Cre-mediated induction of wheat germ agglutinin in transgenic mice
Transneuronal tracing of diverse CNS circuits by Cre-mediated induction of wheat germ agglutinin in transgenic mice
复制标题
DOI：
10.1073/pnas.222546999
复制
发表时间：
2002-11-12
影响因子：
11.1
通讯作者：
Basbaum, AI
中科院分区：
文献类型：
--
作者：
Braz, JM;Rico, B;Basbaum, AI
系统神经科学研究中枢神经系统中神经元群体构成的复杂电路以及这些神经元的协作功能。因此，中枢神经系统回路的神经解剖学分析的改进方法引起了人们的极大兴趣。事实上，最近通过使用在神经元特异性启动子控制下表达跨神经元凝集素示踪剂的转基因小鼠，在纤维束追踪方法方面取得了重大进展。然而，这些动物的用途仅限于受特定启动子影响的中枢神经系统回路。在这里，我们描述了一种新的转基因小鼠，可用于对大脑或脊髓的许多区域的回路进行跨神经元追踪分析。这些小鼠中的转基因导致整个 CNS 神经元中 LacZ 的表达。通过 Cre 介导的重组切除 LacZ 基因会启动凝集素、麦芽凝集素 (WGA) 的表达。为了说明这些 ZW (LacZ-WGA) 小鼠的多种用途，我们通过将小鼠与两个表达 Cre 的转基因小鼠系杂交或通过将表达 Cre 的腺相关病毒显微注射到小脑或大脑皮层来触发 WGA 表达。两种方法均导致发生重组事件的神经元的细胞体和树突中广泛的 WGA 表达，以及凝集素顺行和跨神经元转运至二级和三级神经元。由于凝集素可以在发育中和成年动物以及大脑和脊髓的所有区域中被诱导，因此这些 ZW 对于 CINIS 电路分析的大量研究可能非常有价值。
Systems neuroscience addresses the complex circuits made by populations of neurons in the CNS and the cooperative function of these neurons. Improved approaches to the neuroanatomical analysis of CNS circuits are thus of great interest. In fact, significant advances in tract-tracing methods have recently been made by using transgenic mice that express transneuronal lectin tracers under the control of neuron-specific promoters. The utility of those animals, however, is limited to the CNS circuit influenced by the particular promoter. Here, we describe a new transgenic mouse that can be used for transneuronal tracing analysis of circuits many region of the brain or spinal cord. The transgene in these mice results in expression of LacZ in neurons throughout the CNS. Excision of the LacZ gene by Cre-mediated recombination initiates expression of the lectin, wheat germ agglutinin (WGA). To illustrate the diverse uses of these ZW (LacZ-WGA) mice, we triggered WGA expression either by crossing the mice with two Cre-expressing transgenic mouse lines or by microinjecting a Cre-expressing adeno-associated virus into the cerebellum or cerebral cortex. Both approaches resulted in extensive WGA expression in the cell bodies and dendrites of neurons in which the recombination event occurred, as well as anterograde and transneuronal transport of the lectin to second and third order neurons. Because the lectin can be induced in developing and adult animals, and in all regions of the brain and spinal cord, these ZW may prove extremely valuable for numerous studies of CINIS circuit analysis.