Lehrstuhl für Zellphysiologie, Fakultät für Biologie und Biotechnologie, Ruhr-Universität, Bochum, Germany.
J Neurovirol. 2009 Sep;15(5-6):458-64. doi: 10.3109/13550280903473460.
Pseudorabies virus (PrV) strains such as PrV-Bartha and its marker protein-expressing variants have been used in numerous studies as retrograde transneuronal tracing tools, defining the synaptic organization of mammalian neuronal circuits. However, the possibilities for functional examination of virus-infected neurons are limited to electrophysiological approaches or bulk loading strategies using calcium-sensitive dyes. Herein we report the generation and functional characterization of three PrV-Bartha-derived recombinant virus mutants that express different fluorescent calcium indicator proteins (FCIPs). All three generated virus recombinants are able to infect murine trigeminal neurons and express the corresponding FCIP (GCaMP2, camgaroo-2, or inverse pericam). Functionality of these virally expressed constructs was verified by using confocal Ca-imaging technologies. These FCIP-expressing virus recombinants provide a new tool for the functional analysis of whole circuits of synaptically connected neurons in vitro and in vivo.
伪狂犬病毒（PrV）株，如 PrV-Bartha 及其标记蛋白表达变体，已被广泛用于逆行跨神经元追踪工具的研究中，用于定义哺乳动物神经元回路的突触组织。然而，对于感染病毒的神经元的功能研究仅限于电生理学方法或使用钙敏染料的批量加载策略。在此，我们报告了三种 PrV-Bartha 衍生的重组病毒突变体的产生和功能特征，它们表达不同的荧光钙指示剂蛋白（FCIP）。这三种生成的病毒重组体都能够感染小鼠三叉神经神经元，并表达相应的 FCIP（GCaMP2、camgaroo-2 或反向pericam）。通过使用共聚焦 Ca 成像技术验证了这些病毒表达构建体的功能。这些表达 FCIP 的病毒重组体为体外和体内突触连接神经元全回路的功能分析提供了一种新工具。
