Zhang Xiaonan, Gaudry Quentin
Department of Biology, University of Maryland.
Department of Biology, University of Maryland;
J Vis Exp. 2018 Feb 14(132):57052. doi: 10.3791/57052.
Here, a new technique termed Tetrotoxin (TTX) Engineered Resistance for Probing Synapses (TERPS) is applied to test for monosynaptic connections between target neurons. The method relies on co-expression of a transgenic activator with the tetrodotoxin-resistant sodium channel, NaChBac, in a specific presynaptic neuron. Connections with putative post-synaptic partners are determined by whole-cell recordings in the presence of TTX, which blocks electrical activity in neurons that do not express NaChBac. This approach can be modified to work with any activator or calcium imaging as a reporter of connections. TERPS adds to the growing set of tools available for determining connectivity within networks. However, TERPS is unique in that it also reliably reports bulk or volume transmission and spillover transmission.
在此,一种名为用于探测突触的河豚毒素(TTX)工程抗性(TERPS)的新技术被应用于测试目标神经元之间的单突触连接。该方法依赖于在特定的突触前神经元中,将转基因激活剂与耐河豚毒素的钠通道NaChBac共表达。与假定的突触后伙伴的连接通过在存在河豚毒素的情况下进行全细胞记录来确定,河豚毒素会阻断不表达NaChBac的神经元中的电活动。这种方法可以进行修改,以与任何激活剂配合使用,或使用钙成像作为连接的报告指标。TERPS增加了可用于确定网络内连接性的越来越多的工具集。然而,TERPS的独特之处在于它还能可靠地报告大量或容积传递以及溢出传递。