Braz Joao M, Rico Beatriz, Basbaum Allan I
Department of Anatomy and Physiology and W. M. Keck Foundation Center for Integrative Neuroscience, University of California, San Francisco 94143, USA.
Proc Natl Acad Sci U S A. 2002 Nov 12;99(23):15148-53. doi: 10.1073/pnas.222546999. Epub 2002 Oct 21.
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 in any 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 CNS circuit analysis.
系统神经科学研究中枢神经系统中神经元群体形成的复杂回路以及这些神经元的协同功能。因此,改进中枢神经系统回路的神经解剖学分析方法备受关注。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,在示踪方法上取得了重大进展。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。在此,我们描述了一种新的转基因小鼠,可用于对大脑或脊髓任何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LacZ在整个中枢神经系统的神经元中表达。通过Cre介导的重组切除LacZ基因会启动凝集素小麦胚芽凝集素(WGA)的表达。为了说明这些ZW(LacZ-WGA)小鼠的多种用途,我们通过将这些小鼠与两种表达Cre的转基因小鼠品系杂交,或通过将表达Cre的腺相关病毒微量注射到小脑或大脑皮层中来触发WGA表达。两种方法都导致在发生重组事件的神经元的细胞体和树突中广泛表达WGA,以及凝集素向二级和三级神经元的顺行和跨神经元运输。由于凝集素可在发育中和成年动物以及大脑和脊髓的所有区域中诱导产生,这些ZW小鼠可能对中枢神经系统回路分析的众多研究极具价值。