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An R-CaMP1.07 reporter mouse for cell-type-specific expression of a sensitive red fluorescent calcium indicator

机译:R-CaMP1.07报告基因小鼠,用于敏感的红色荧光钙指示剂的细胞类型特异性表达

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摘要

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.
机译:遗传编码的钙指示剂(GECI)能够以高灵敏度和特异性对体内脑细胞活性进行成像。与病毒感染或子宫内电穿孔相反,转基因报告基因系中的指示剂表达是非侵入性,可靠且均质的。最近,引入了Cre / tTA依赖性报告基因小鼠,当与Cre和tTA驱动程序小鼠杂交时,它们以细胞类型特异性和可诱导的方式提供绿色荧光GECI的高水平表达。在这里,我们使用R-CaMP1.07生成并表征了该类型的第一条红移GECI报告基因系,R-CaMP1.07是一种红色荧光指示剂,可以有效地在1000 nm以上被双光子激发。通过将新的R-CaMP1.07报告基因谱系与驱动新皮质中层特异性表达的Cre谱系交叉,我们证明了其高度真实性,可报告体内潜在的动作电位触发,数月的长期稳定性以及对兴奋性神经元功能成像的广泛用途跨所有皮质层,尤其是在以前难以接近的第4和第6层中。

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