In vivo calcium imaging from genetically specified target cells in mouse cerebellum
In vivo calcium imaging from genetically specified target cells in mouse cerebellum
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DOI：
10.1016/j.neuroimage.2006.10.021
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发表时间：
2007-02-01
期刊：
影响因子：
5.7
通讯作者：
Knopfel, Thomas
中科院分区：
文献类型：
--
作者：
Diez-Garcia, Javier;Akemann, Walther;Knopfel, Thomas
基因编码的荧光钙指示蛋白提供了监测基因指定靶细胞活性的潜力，而不需要单细胞分辨率。在这里，我们报道了在小脑颗粒细胞中表达荧光钙指示蛋白GCaMP2的转基因小鼠在体内经颅成像平行纤维活性。我们展示了通过完整颅骨，平行纤维束响应电刺激在分子层的钙瞬态的可靠测量。这些平行纤维钙瞬态与固有的突触后自身荧光信号的不同之处在于其更快的动力学和对突触传递阻滞剂的抗性。最后，我们使用2P激光扫描显微镜来证明在高空间分辨率下，平行纤维束对活小鼠钙瞬态的可靠测量。我们期望基因靶向的荧光钙指示蛋白与光学成像技术将有助于构建特定脑回路功能的宏观和微观图谱。(c) 2006爱思唯尔公司版权所有。
Genetically encoded fluorescent calcium indicator proteins provide the potential to monitor activity from genetically specified target cells without a need for single cell resolution. Here we report the use of transgenic mice expressing the fluorescent calcium indicator protein GCaMP2 in cerebellar granule cells to image parallel fiber activity transcranially in vivo. We demonstrated reliable measurements of calcium transients from beams of parallel fibers in response to electrical stimulation in the molecular layer through the intact skull. These parallel fiber calcium transients differed from intrinsic postsynaptic autofluorescence signals in their faster kinetics and resistance to blockers of synaptic transmission. Finally, we used 2P laser-scanning microscopy to demonstrate reliable measurements of calcium transients from beams of parallel fibers at high spatial resolution in living mice. We expect that genetically targeted fluorescent calcium indicator proteins along with optical imaging techniques will be instrumental for the construction of macroscopic and microscopic maps of the function of specific brain circuits. (c) 2006 Elsevier Inc. All rights reserved.