Use of pseudorabies virus to delineate multisynaptic circuits in brain: opportunities and limitations
Use of pseudorabies virus to delineate multisynaptic circuits in brain: opportunities and limitations
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DOI：
10.1016/s0165-0270(00)00295-8
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发表时间：
2000-11-15
影响因子：
3
通讯作者：
Card, JP
中科院分区：
文献类型：
--
作者：
Aston-Jones, G;Card, JP
活病毒跨突触追踪是一种强大的工具，已被广泛用于分析调节外周靶点的中枢传出神经。最近，研究人员开始使用这种新的方法，通过中央注射来识别大脑内的电路解剖。虽然外周注射伪狂犬病病毒的跨突触示踪已经被广泛描述，但与这种示踪剂的中枢应用相关的几个方法学问题还没有得到解决。在这里，我们回顾了在大鼠脑内注射病毒的实验中与使用伪狂犬病病毒(PRV；Bartha株)相关的下列问题：(I)决定病毒摄取区域的因素；(Ii)传代纤维对伪狂犬病病毒的摄取；(Iii)病毒渗入脑室后病毒对大脑的入侵；(Iv)考虑将PRV与多肽和神经递质双重标记；(V)使用PRV与传统逆行示踪剂一起在解剖学上识别多突触通路中的中继；以及(Vi)将PRV在树突树中的运输作为识别远端树突输入的一种手段。总而言之，这些数据表明，当适当的控制被纳入实验设计时，PRV为解剖中枢神经系统的突触提供了一种强大的手段。本文末尾提供了一组用于各种过程的方法。(C)2000 Elsevier Science B.V.保留所有权利。
Transsynaptic tracing with live virus is a powerful tool that has been used extensively to analyze central efferents that regulate peripheral targets. More recently, investigators have begun to use this new methodology with central injections to identify circuit anatomy within the brain. Although transsynaptic tracing with peripheral injection of pseudorabies virus has been extensively characterized, several methodological issues related to central application of this tracer have not been addressed. Here, we review the following issues relevant to the use of pseudorabies virus (PRV; Bartha strain) in experiments involving injection of virus into rat brain: (i) factors that determine the zone of viral uptake; (ii) uptake of pseudorabies virus by fibers of passage; (iii) viral invasion of the brain after leakage of virus into the brain ventricles; (iv) considerations for double labeling for PRV with peptides and neurotransmitters; (v) use of PRV with conventional retrograde tracers to anatomically identify relays in a multisynaptic pathway; and (vi) transport of PRV throughout the dendritic tree as a means of identifying inputs to distal dendrites. Collectively, the data demonstrate that PRV provides a powerful means of dissecting the synaptology of CNS circuitry when appropriate controls are incorporated into the experimental design. A set of recipes for various procedures are included at the end of this article. (C) 2000 Elsevier Science B.V. All rights reserved.