Imaging high-resolution structure of GFP-expressing neurons in neocortex in vivo
Imaging high-resolution structure of GFP-expressing neurons in neocortex in vivo
复制标题
DOI：
10.1101/lm.32700
复制
发表时间：
2000-11-01
影响因子：
2
通讯作者：
Svoboda, K
中科院分区：
文献类型：
--
作者：
Chen, BE;Lendvai, B;Svoboda, K
为了检测神经元形态学响应于经验变化的细微变化，必须在几分钟到几天的时间尺度内以高分辨率对体内神经元进行成像。我们通过用携带增强型绿色荧光蛋白基因的辛德毕斯病毒感染大鼠和小鼠桶状皮层的有丝分裂后神经元来实现这一点。用双光子激发激光扫描显微镜观察，感染的神经元显示出均匀分布在整个细胞中的明亮荧光，包括轴突和树突状乔木。单树突棘可以常规解决和可视化的形态动力学。病毒感染和成像在整个出生后发育直至成年早期（P8 -30）实现，尽管病毒感染效率随年龄而降低。这种相对非侵入性的荧光标记和成像的神经元的方法允许在体内的新皮层神经元及其电路的形态动力学的研究。
To detect subtle changes in neuronal morphology in response to changes in experience, one must image neurons at high resolution in vivo over time scales of minutes to days. We accomplished this by infecting postmitotic neurons in rat and mouse barrel cortex with a Sindbis virus carrying the gene for enhanced green fluorescent protein. Visualized with 2-photon excitation laser scanning microscopy, infected neurons showed bright fluorescence that was distributed homogeneously throughout the cell, including axonal and dendritic arbors. Single dendritic spines could routinely be resolved and their morphological dynamics visualized. Viral infection and imaging were achieved throughout postnatal development up to early adulthood (P 8-30), although the viral efficiency of infection decreased with age. This relatively noninvasive method for fluorescent labeling and imaging of neurons allows the study of morphological dynamics of neocortical neurons and their circuits in vivo.