Max Planck Institute of Neurobiology, Martinsried, Germany.
Graduate School of Systemic Neurosciences, LMU Munich, Martinsried, Germany.
Elife. 2020 Oct 20;9:e62953. doi: 10.7554/eLife.62953.
Neurotransmitter receptors and ion channels shape the biophysical properties of neurons, from the sign of the response mediated by neurotransmitter receptors to the dynamics shaped by voltage-gated ion channels. Therefore, knowing the localizations and types of receptors and channels present in neurons is fundamental to our understanding of neural computation. Here, we developed two approaches to visualize the subcellular localization of specific proteins in : The flippase-dependent expression of GFP-tagged receptor subunits in single neurons and 'FlpTag', a versatile new tool for the conditional labelling of endogenous proteins. Using these methods, we investigated the subcellular distribution of the receptors GluClα, Rdl, and Dα7 and the ion channels para and Ih in motion-sensing T4/T5 neurons of the visual system. We discovered a strictly segregated subcellular distribution of these proteins and a sequential spatial arrangement of glutamate, acetylcholine, and GABA receptors along the dendrite that matched the previously reported EM-reconstructed synapse distributions.
神经递质受体和离子通道塑造了神经元的生物物理特性，从神经递质受体介导的反应的特征到电压门控离子通道塑造的动力学。因此，了解神经元中存在的受体和通道的定位和类型是我们理解神经计算的基础。在这里，我们开发了两种方法来可视化特定蛋白质在：翻转酶依赖性 GFP 标记的受体亚基在单个神经元中的表达和“FlpTag”，一种用于内源性蛋白质条件标记的新的多功能工具。使用这些方法，我们研究了运动感应 T4/T5 神经元中 GluClα、Rdl 和 Dα7 受体以及 para 和 Ih 离子通道的亚细胞分布。我们发现这些蛋白质的严格分隔的亚细胞分布以及谷氨酸、乙酰胆碱和 GABA 受体沿着树突的顺序空间排列与先前报道的 EM 重建的突触分布相匹配。
