Visualization of cortico-basal ganglia loop by means of transsynaptic transport of WGA-recombinant adeno virus and rabies virus

利用wga重组腺病毒和狂犬病毒的跨突触转运进行逆行跨神经元标记，试图分析与功能相关的多突触通路的组织，例如，来自不同额叶运动区和关联区的皮质-基底节区环路。在本报告中，我们只描述了从狂犬病毒的一系列实验中获得的数据，因为在本研究中发现狂犬病毒比wga -重组腺病毒更敏感。将狂犬病毒（CVS-11； CDC）注射到初级运动皮层（MI），以研究其在中央运动系统中的逆行运输程度。将病毒悬浮液注射到经电生理学鉴定的恒河猴心肌梗死前肢区域。注射后2至4天，猴子进行灌注固定。连续冠状切片进行狂犬病毒免疫组化染色。注射病毒三天后，许多二级神经元被标记，以及丘脑和大脑皮层直接投射到心梗的一级神经元。在基底节区，在苍白球内段的腹侧观察到标记神经元。此外，神经元标记发生在小脑深部核，特别是外侧核和位间核的背侧部分，以及额叶几个位于背侧的运动相关区域。注射后三天半左右，在壳核腹侧、丘脑底核、白球外段，以及小脑皮层和前额叶皮层的浦肯野细胞中，可以看到三级神经元。此外，我们的免疫荧光分析证实，在注射后4天，胆碱能纹状体中间神经元和多巴胺能神经元（可能是基底节区的四阶神经元）被标记。少

Retrograde transneuronal labeling by means of transsynaptic transport of WGA-recombinant adeno virus and rabies virus was attempted to analyze the organization of functionally-related multi-synaptic pathways, for example, the cortico-basal ganglia loops that arise from various frontal motor and association areas. In this report, we have only described the data obtained from a series of experiments using rabies virus, because rabies virus was found to be much more sensitive than WGA-recombinant adeno virus for the present purpose. Injections of rabies virus (CVS-11; CDC) were made into the primary motor cortex (MI) to investigate the extent of its retrograde transport in the central motor system. A viral suspension was injected into the electrophysiologically identified forelimb region of the MI in rhesus monkeys. The monkeys underwent perfusion-fixation two to four days after the injection. Serial coronal sections were processed for immunohistochemical staining for rabies virus. Three … More days after viral injection, many second-order neurons were labeled, along with the first-order neurons in the thalamus and the cerebral cortex that project to the MI directly. In the basal ganglia, the labeled neurons were observed in the ventral aspect of the internal segment of the globus pallidus. Moreover, neuronal labeling occurred in the deep cerebellar nuclei, especially in the dorsal portions of the lateral and interpositus nuclei, and in several rostrally-situated motor-related areas of the frontal lobe. Around three and a half days after the injection, third-order neurons were seen in the ventral aspects of the putamen, the subthalamic nucleus, and the external segment of the globus pallidus, as well as in Purkinje cells of the cerebellar cortex and areas of the prefrontal cortex. In addition, our immunofluorescence analysis confirmed that cholinergic striatal interneurons and dopaminergic nigral neurons, probably fourth-order neurons in the basal ganglia, were labeled at the four-day postinjection period. Less