An R-CaMP1.07 reporter mouse for cell-type-specific expression of a sensitive red fluorescent calcium indicator.
An R-CaMP1.07 reporter mouse for cell-type-specific expression of a sensitive red fluorescent calcium indicator.
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DOI：
10.1371/journal.pone.0179460
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Helmchen F
中科院分区：
文献类型：
--
作者：
Bethge P;Carta S;Lorenzo DA;Egolf L;Goniotaki D;Madisen L;Voigt FF;Chen JL;Schneider B;Ohkura M;Nakai J;Zeng H;Aguzzi A;Helmchen F
遗传编码钙指标（GECIs）能够以高灵敏度和特异性成像体内脑细胞活动。与病毒感染或子宫内电穿孔相比，转基因报告系中的指示物表达是无创、可靠和均匀的。最近，引入了Cre/tTA依赖的报告小鼠，与Cre和tTA驱动小鼠杂交后，绿色荧光GECIs以细胞类型特异性和诱导的方式高水平表达。在这里，我们使用R-CaMP1.07生成并表征了这种类型的第一个红移GECI报告线，R-CaMP1.07是一种在1000 nm以上有效激发双光子的红色荧光指示器。通过将新的R-CaMP1.07报告线与驱动新皮层层特异性表达的Cre线交叉，我们证明了其在体内报告动作电位放电的高保真度，数月的长期稳定性，以及在所有皮层层的兴奋性神经元功能成像的多功能，特别是在以前难以访问的第4层和第6层。
Genetically encoded calcium indicators (GECIs) enable imaging of in vivo brain cell activity with high sensitivity and specificity. In contrast to viral infection or in utero electroporation, indicator expression in transgenic reporter lines is induced noninvasively, reliably, and homogenously. Recently, Cre/tTA-dependent reporter mice were introduced, which provide high-level expression of green fluorescent GECIs in a cell-type-specific and inducible manner when crossed with Cre and tTA driver mice. Here, we generated and characterized the first red-shifted GECI reporter line of this type using R-CaMP1.07, a red fluorescent indicator that is efficiently two-photon excited above 1000 nm. By crossing the new R-CaMP1.07 reporter line to Cre lines driving layer-specific expression in neocortex we demonstrate its high fidelity for reporting action potential firing in vivo, long-term stability over months, and versatile use for functional imaging of excitatory neurons across all cortical layers, especially in the previously difficult to access layers 4 and 6.