Imaging activity in neurons and glia with a Polr2a-based and cre-dependent GCaMP5G-IRES-tdTomato reporter mouse.
Imaging activity in neurons and glia with a Polr2a-based and cre-dependent GCaMP5G-IRES-tdTomato reporter mouse.
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DOI：
10.1016/j.neuron.2014.07.024
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发表时间：
2014-09-03
期刊：
影响因子：
16.2
通讯作者：
Tvrdik P
中科院分区：
文献类型：
--
作者：
Gee JM;Smith NA;Fernandez FR;Economo MN;Brunert D;Rothermel M;Morris SC;Talbot A;Palumbos S;Ichida JM;Shepherd JD;West PJ;Wachowiak M;Capecchi MR;Wilcox KS;White JA;Tvrdik P
将遗传编码的活动指标引入动物模型的新策略有助于神经系统功能的研究。我们已经开发了PC：：G5-tdT小鼠系，其以Cre依赖性方式表达GCaMP 5G钙指示剂。我们没有靶向ROSA 26基因座，而是将报告基因盒插入到普遍表达的Polr 2a基因附近，而不破坏基因座的完整性。用IRES-tdTomato标记指示剂，以帮助检测阳性细胞。该报告系统在广泛的发育和细胞环境中是有效的。我们记录了完整清醒的新生儿自发皮质钙波，并评估了浓度依赖性反应的气味在成人嗅球。此外，PC：：G5-tdT有效地报告星形胶质细胞和小胶质细胞的胞体和精细过程中的细胞内钙动力学。通过电生理和行为分析，我们确定GCaMP 5G表达对神经系统性能没有重大影响。PC：：G5-tdT将用于各种脑映射实验。
New strategies for introducing genetically encoded activity indicators into animal models facilitate the investigation of nervous system function. We have developed the PC::G5-tdT mouse line that expresses the GCaMP5G calcium indicator in a Cre-dependent fashion. Instead of targeting the ROSA26 locus, we inserted the reporter cassette nearby the ubiquitously expressed Polr2a gene without disrupting locus integrity. The indicator was tagged with IRES-tdTomato to aid detection of positive cells. This reporter system is effective in a wide range of developmental and cellular contexts. We recorded spontaneous cortical calcium waves in intact awake newborns and evaluated concentration-dependent responses to odorants in the adult olfactory bulb. Moreover, PC::G5-tdT effectively reports intracellular calcium dynamics in somas and fine processes of astrocytes and microglial cells. Through electrophysiological and behavioral analyses, we determined that GCaMP5G expression had no major impact on nervous system performance. PC::G5-tdT will be instrumental for a variety of brain mapping experiments.
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DOI：
10.1073/pnas.250471697
发表时间：
2000-12-05
影响因子：
11.1
作者：
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通讯作者：
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影响因子：
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Feng G
影响因子：
25
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