Imaging neuronal activity in the central and peripheral nervous systems using new Thy1.2-GCaMP6 transgenic mouse lines
Imaging neuronal activity in the central and peripheral nervous systems using new Thy1.2-GCaMP6 transgenic mouse lines
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DOI：
10.1016/j.jneumeth.2019.108535
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发表时间：
2020-03-15
影响因子：
3
通讯作者：
Gan, Wen-Biao
中科院分区：
文献类型：
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作者：
Cichon, Joseph;Magrane, Jordi;Gan, Wen-Biao
背景资料：基因编码的钙（Ca 2+）传感器GCaMP 6已被广泛用于神经元胞体、树突和突触中钙瞬变的成像。新方法：在此我们描述了五种新的转基因小鼠品系，它们在Thy1.2启动子的控制下在中枢和外周神经系统中表达GCaMP 6 F（快）或GCaMP 6S（慢）。这些转基因株系在多个脑区域中表现出GCaMP 6的稳定和层特异性表达。与现有的Thy1.2-GCaMP 6小鼠相比，它们具有几个独特的特征，包括GCaMP 6在大脑皮层的第V层锥体神经元、脊髓中的运动神经元以及背根神经节（DRG）中的感觉神经元中的稀疏表达。我们进一步证明，这些小鼠线允许在麻醉和清醒行为小鼠的大脑皮层中的神经元胞体和顶端树突以及DRG神经元中的Ca 2+瞬变的稳健检测。与现有的方法比较：这些转基因株系允许钙离子成像的树突和胞体的锥体神经元在特定的皮质layers.Conclusions：这些GCaMP 6转基因株系，从而提供了有用的工具，在中枢和外周神经系统的神经元回路的功能分析。
Background: The genetically encoded calcium (Ca2+) sensor GCaMP6 has been widely used for imaging Ca2+ transients in neuronal somata, dendrites, and synapses.New method: Here we describe five new transgenic mouse lines expressing GCaMP6F (fast) or GCaMP6S (slow) in the central and peripheral nervous system under the control of theThy1.2 promoter.Results: These transgenic lines exhibit stable and layer-specific expression of GCaMP6 in multiple brain regions. They have several unique features compared to existing Thy1.2-GCaMP6 mice, including sparse expression of GCaMP6 in layer V pyramidal neurons of the cerebral cortex, motor neurons in the spinal cord, as well as sensory neurons in dorsal root ganglia (DRG). We further demonstrate that these mouse lines allow for robust detection of Ca2+ transients in neuronal somata and apical dendrites in the cerebral cortex of both anesthetized and awake behaving mice, as well as in DRG neurons. Comparison with existing method(s): These transgenic lines allows Ca2+ imaging of dendrites and somas of pyramidal neurons in specific cortical layers that is difficult to achieve with existing methods.Conclusions: These GCaMP6 transgenic lines thus provide useful tools for functional analysis of neuronal circuits in both central and peripheral nervous systems.