Sensitive red protein calcium indicators for imaging neural activity
Sensitive red protein calcium indicators for imaging neural activity
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DOI：
10.7554/elife.12727
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发表时间：
2016-03-24
期刊：
影响因子：
7.7
通讯作者：
Kim, Douglas S.
中科院分区：
文献类型：
--
作者：
Dana, Hod;Mohar, Boaz;Kim, Douglas S.
基因编码的钙指示器(GEC)允许测量大量神经元和小神经元隔间的活动，时间从几毫秒到几个月。尽管基于GFP的GEC被广泛用于体内神经生理学，但具有红移激发和发射光谱的GEC在体内成像方面具有优势，因为它减少了组织中的散射和吸收，从而降低了光毒性。然而，当前的红色GEC在检测和量化神经活动方面不如最先进的基于GFP的GCaMP6指示器。在这里，我们提出了基于mRuby(jRCaMP1a，b)和MApple(JRGECO1a)的改进的红色GEC，其灵敏度与GCaMP6相当。我们在培养的神经元以及小鼠、果蝇、斑马鱼和线虫体内表征了新的红色GEC的性能。红色GEC促进了深层组织成像、与基于GFP的记者一起的双色成像，以及光遗传学与钙成像的结合使用。
Genetically encoded calcium indicators (GEC's) allow measurement of activity in large populations of neurons and in small neuronal compartments, over times of milliseconds to months. Although GFP-based GEC's are widely used for in vivo neurophysiology, GEC's with red-shifted excitation and emission spectra have advantages for in vivo imaging because of reduced scattering and absorption in tissue, and a consequent reduction in phototoxicity. However, current red GEC's are inferior to the state-of-the-art GFP-based GCaMP6 indicators for detecting and quantifying neural activity. Here we present improved red GEC's based on mRuby (jRCaMP1a, b) and mApple (jRGECO1a), with sensitivity comparable to GCaMP6. We characterized the performance of the new red GEC's in cultured neurons and in mouse, Drosophila, zebrafish and C. elegans in vivo. Red GEC's facilitate deep-tissue imaging, dual-color imaging together with GFP-based reporters, and the use of optogenetics in combination with calcium imaging.