In Vivo Imaging and Noninvasive Ablation of Pyramidal Neurons in Adult NEX-CreERT2 Mice
In Vivo Imaging and Noninvasive Ablation of Pyramidal Neurons in Adult NEX-CreERT2 Mice
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DOI：
10.1093/cercor/bhr214
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发表时间：
2012-07-01
期刊：
影响因子：
3.7
通讯作者：
Schwab, Markus H.
中科院分区：
文献类型：
--
作者：
Agarwal, Amit;Dibaj, Payam;Schwab, Markus H.
在小鼠中研究嵌入复杂神经网络中的单个神经元的功能是困难的。条件诱变允许基因表达的时空控制，包括毒素对细胞的消融。直接表达他莫昔芬诱导的Cre重组酶（CreERT 2）的变体选择性皮质神经元，我们取代了小鼠Nex 1基因的编码区CreERT 2 cDNA通过同源重组在胚胎干细胞。当注射他莫昔芬时，成年NEX-CreERT 2小鼠仅在新皮质和海马的投射神经元中诱导报告基因表达。通过滴定他莫昔芬的剂量，我们实现了单细胞重组，这使得多光子成像的新皮层神经元在活的小鼠。当海马投射神经元通过白喉毒素诱导表达进行基因消融时，在20天内炎症反应包括CD 3 + T细胞的浸润。这标志着与类似研究的显著差异，在类似研究中，垂死的少突胶质细胞无法招募适应性免疫系统的细胞。
To study the function of individual neurons that are embedded in a complex neural network is difficult in mice. Conditional mutagenesis permits the spatiotemporal control of gene expression including the ablation of cells by toxins. To direct expression of a tamoxifen-inducible variant of Cre recombinase (CreERT2) selectively to cortical neurons, we replaced the coding region of the murine Nex1 gene by CreERT2 cDNA via homologous recombination in embryonic stem cells. When injected with tamoxifen, adult NEX-CreERT2 mice induced reporter gene expression exclusively in projection neurons of the neocortex and hippocampus. By titrating the tamoxifen dosage, we achieved recombination in single cells, which allowed multiphoton imaging of neocortical neurons in live mice. When hippocampal projection neurons were genetically ablated by induced expression of diphteria toxin, within 20 days the inflammatory response included the infiltration of CD3+ T cells. This marks a striking difference from similar studies, in which dying oligodendrocytes failed to recruit cells of the adaptive immune system.