A genetically encoded calcium indicator for chronic in vivo two-photon imaging
A genetically encoded calcium indicator for chronic in vivo two-photon imaging
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DOI：
10.1038/nmeth.1243
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发表时间：
2008-09-01
期刊：
影响因子：
48
通讯作者：
Griesbeck, Oliver
中科院分区：
文献类型：
--
作者：
Mank, Marco;Santos, Alexandre Ferrao;Griesbeck, Oliver
神经系统中的神经元可以随着时间的推移改变其功能特性。目前，还没有技术可以在重复的实验会话中可靠地监测所识别的神经元内的变化。我们通过肌钙蛋白C钙结合部分内的诱变和结构域重排来增加基于肌钙蛋白C的钙生物传感器在低钙状态下的信号强度，以产生指示剂TN-XXL。使用体内双光子比率成像，我们表明，TN-XXL表现出增强的荧光变化，在苍蝇和小鼠的神经元。TN-XXL可用于获得小鼠视皮层方向选择性神经元的调谐曲线，该曲线在几天和几周内重复测量。因此，基因编码的钙指示剂TN-XXL允许重复成像的响应特性从个别，确定神经元在体内，这将是至关重要的获得新的见解可塑性，再生和疾病的细胞机制。
Neurons in the nervous system can change their functional properties over time. At present, there are no techniques that allow reliable monitoring of changes within identified neurons over repeated experimental sessions. We increased the signal strength of troponin C-based calcium biosensors in the low-calcium regime by mutagenesis and domain rearrangement within the troponin C calcium binding moiety to generate the indicator TN-XXL. Using in vivo two-photon ratiometric imaging, we show that TN-XXL exhibits enhanced fluorescence changes in neurons of flies and mice. TN-XXL could be used to obtain tuning curves of orientation-selective neurons in mouse visual cortex measured repeatedly over days and weeks. Thus, the genetically encoded calcium indicator TN-XXL allows repeated imaging of response properties from individual, identified neurons in vivo, which will be crucial for gaining new insights into cellular mechanisms of plasticity, regeneration and disease.