Transneuronal circuit analysis with pseudorabies viruses.
Transneuronal circuit analysis with pseudorabies viruses.
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DOI：
10.1002/0471142301.ns0105s09
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发表时间：
2001-05-01
影响因子：
--
通讯作者：
Enquist, L W
中科院分区：
文献类型：
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作者：
Card, J P;Enquist, L W
在过去的十年中，病毒作为神经元回路的跨神经元示踪剂的使用急剧增加。该方法利用嗜神经病毒侵入神经元的倾向，然后产生感染性后代，这些后代穿过突触感染回路中的其他神经元。包括在这个单位的协议和评论集中在使用猪α疱疹病毒称为伪狂犬病病毒（PRV）的多突触分析。在这里，方面的实验设计，有最大的进口成功使用病毒的电路定义。因此，本单元中包含的协议可以与经典道示踪剂的使用方法一起应用。详细介绍了通过外周注射病毒逆行感染大鼠CNS回路的程序，同时还描述了通过脑内注射进行的transneuronal分析。这些程序的变体，多个重组株的跨神经元分析，也被描述沿着与用于生长和滴定病毒储备的方法，以及用于在固定的脑组织中病毒抗原的单和双免疫组织化学定位的程序。
Over the past decade there has been a dramatic increase in the use of viruses as transneuronal tracers of neuronal circuitry. The method exploits the propensity of neurotropic viruses to invade neurons and then produce infectious progeny that cross synapses to infect other neurons within a circuit. The protocols and commentaries included in this unit focus upon the use of the swine alpha herpesvirus known as pseudorabies virus (PRV) for polysynaptic analysis. Here, the aspects of experimental design that have the greatest import for successful use of viruses in circuit definition are presented. Accordingly, the protocols included in this unit can be applied in concert with methods in which the use of classical tract tracers has been detailed. A procedure for retrograde infection of CNS circuits in the rat CNS by peripheral injection of virus is detailed, while transneuronal analysis by intracerebral injection is also described. A variant of these procedures, transneuronal analysis with multiple recombinant strains, is also described along with methods for growing and titering viral stocks, and procedures for single and dual immunohistochemical localization of viral antigens in fixed brain tissue.