In vivo imaging of neuronal activity - Neurotechnique by targeted expression of a genetically encoded probe in the mouse
In vivo imaging of neuronal activity - Neurotechnique by targeted expression of a genetically encoded probe in the mouse
复制标题
DOI：
10.1016/s0896-6273(04)00144-8
复制
发表时间：
2004-04-08
期刊：
影响因子：
16.2
通讯作者：
Wachowiak, M
中科院分区：
文献类型：
--
作者：
Bozza, T;McGann, JP;Wachowiak, M
基因编码探针在对完整神经系统中特定神经元群进行功能成像方面显示出巨大的前景，但它们的体内应用受到限制。在这里，我们将突触-pHluorin（一种报告突触小泡融合的 pH 敏感蛋白）靶向表达到小鼠的嗅觉感觉神经元。 Synapto-pHluorin 选择性标记嗅球肾小球中感觉神经元的突触前末梢。气味刺激引起大幅荧光增加，该荧光增加局限于体内单个肾小球，与突触前钙流入相关，根据刺激强度分级，并在几天内保持稳定。气味激活的肾小球的空间模式是分布的，并且不会随着碳链长度的增加而系统地改变，这与使用其他成像方法报道的精细组织的化学性质相反。突触-pHluorin 在小鼠中的靶向表达将允许分析以前无法访问的神经元群体，并从体内基因鉴定的神经元进行长期成像。
Genetically encoded probes show great promise in permitting functional imaging of specified neuronal populations in the intact nervous system, yet their in vivo application has been limited. Here, we have targeted expression of synapto-pHluorin, a pH-sensitive protein that reports synaptic vesicle fusion, to olfactory sensory neurons in mouse. Synapto-pHluorin selectively labeled presynaptic terminals of sensory neurons in glomeruli of the olfactory bulb. Odorant stimulation evoked large-amplitude fluorescence increases that were localized to individual glomeruli in vivo, correlated with presynaptic calcium influx, graded with stimulus intensity, and stable over a period of days. Spatial patterns of odorant-activated glomeruli were distributed and did not change systematically with increasing carbon chain length, in contrast to the finely organized chemotopy that has been reported using other imaging methods. Targeted expression of synapto-pHluorin in mouse will permit the analysis of previously inaccessible neuronal populations and chronic imaging from genetically identified neurons in vivo.