Tabuchi K, Sawamoto K, Suzuki E, Ozaki K, Sone M, Hama C, Tanifuji-Morimoto T, Yuasa Y, Yoshihara Y, Nose A, Okano H
Department of Neuroscience, Biomedical Research Center, Osaka University Graduate School of Medicine, Suita, Japan.
J Neurosci Res. 2000 Jan 1;59(1):94-9.
Visualization of specific transsynaptic neural pathways is an indispensable technique for understanding the relationship between structure and function in the nervous system. Here, we demonstrate the application of the wheat germ agglutinin (WGA) transgene technique for tracing transsynaptic neural pathways in Drosophila. The intracellular localization of WGA was examined by immunoelectron microscopy. WGA signals were detected in granule-like structures in both the outer photoreceptor cells expressing WGA and the second-order laminar neurons. Misexpression of tetanus toxin (TNT), which inactivates N-synaptobrevin, in the outer photoreceptor cells resulted in the elimination of on/off transients in electroretinogram (ERG) recordings and in a great reduction in WGA transfer into laminar neurons, suggesting that anterograde WGA transsynaptic transfer is dependent mainly on synaptic transmission. Retrograde WGA transfer was also detected upon its forced expression in muscle cells. WGA primarily expressed in muscle cells was taken up by motoneuron axons and transported to their cell bodies in the ventral nerve cord, suggesting that WGA can trace motoneuronal pathways in combination with the muscle-specific GAL4 driver. Thus, the GAL4/UAS-WGA system should facilitate the dissection of the Drosophila neural circuit formation and/or synaptic activity in various regions and at various developmental stages.
可视化特定的跨突触神经通路是理解神经系统结构与功能关系的一项不可或缺的技术。在此,我们展示了小麦胚凝集素(WGA)转基因技术在果蝇中追踪跨突触神经通路的应用。通过免疫电子显微镜检查了WGA的细胞内定位。在表达WGA的外感光细胞和二级层状神经元的颗粒状结构中均检测到WGA信号。在外感光细胞中错误表达使N - 突触囊泡蛋白失活的破伤风毒素(TNT),导致视网膜电图(ERG)记录中的开/关瞬态消失,并且WGA向层状神经元的转移大幅减少,这表明顺行性WGA跨突触转移主要依赖于突触传递。在肌肉细胞中强制表达WGA时也检测到了逆行性WGA转移。主要在肌肉细胞中表达的WGA被运动神经元轴突摄取并运输到腹神经索中的细胞体,这表明WGA可以与肌肉特异性GAL4驱动蛋白结合追踪运动神经元通路。因此,GAL4/UAS - WGA系统应有助于剖析果蝇在各个区域和不同发育阶段的神经回路形成和/或突触活动。