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用于体内成像的改良基因编码近红外荧光钙离子指示剂。

Improved genetically encoded near-infrared fluorescent calcium ion indicators for in vivo imaging.

机构信息

Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada.

Departments of Biological Engineering, Media Arts and Sciences, Brain and Cognitive Sciences, McGovern Institute, Koch Institute, Center for Neurobiological Engineering, MIT, and Howard Hughes Medical Institute, Cambridge, Massachusetts, United States of America.

出版信息

PLoS Biol. 2020 Nov 24;18(11):e3000965. doi: 10.1371/journal.pbio.3000965. eCollection 2020 Nov.

DOI:10.1371/journal.pbio.3000965
PMID:33232322
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7723245/
Abstract

Near-infrared (NIR) genetically encoded calcium ion (Ca2+) indicators (GECIs) can provide advantages over visible wavelength fluorescent GECIs in terms of reduced phototoxicity, minimal spectral cross talk with visible light excitable optogenetic tools and fluorescent probes, and decreased scattering and absorption in mammalian tissues. Our previously reported NIR GECI, NIR-GECO1, has these advantages but also has several disadvantages including lower brightness and limited fluorescence response compared to state-of-the-art visible wavelength GECIs, when used for imaging of neuronal activity. Here, we report 2 improved NIR GECI variants, designated NIR-GECO2 and NIR-GECO2G, derived from NIR-GECO1. We characterized the performance of the new NIR GECIs in cultured cells, acute mouse brain slices, and Caenorhabditis elegans and Xenopus laevis in vivo. Our results demonstrate that NIR-GECO2 and NIR-GECO2G provide substantial improvements over NIR-GECO1 for imaging of neuronal Ca2+ dynamics.

摘要

近红外（NIR）基因编码钙离子（Ca2+）指示剂（GECIs）在减少光毒性、与可见光激发的光遗传学工具和荧光探针的最小光谱串扰，以及减少哺乳动物组织中的散射和吸收方面，相对于可见波长荧光 GECIs 具有优势。我们之前报道的 NIR GECI，NIR-GECO1，具有这些优势，但与最先进的可见波长 GECIs 相比，在用于神经元活动成像时，其亮度较低，荧光响应有限，也存在一些缺点。在这里，我们报告了 2 种改进的 NIR GECI 变体，分别命名为 NIR-GECO2 和 NIR-GECO2G，源自 NIR-GECO1。我们在培养细胞、急性小鼠脑切片以及活体秀丽隐杆线虫和非洲爪蟾中对新的 NIR GECI 的性能进行了表征。我们的结果表明，NIR-GECO2 和 NIR-GECO2G 为神经元 Ca2+动力学成像提供了比 NIR-GECO1 显著的改进。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb49/7723245/858ea2786fe4/pbio.3000965.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb49/7723245/59cf9999817c/pbio.3000965.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb49/7723245/a596eaa039f4/pbio.3000965.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb49/7723245/2707f3959e35/pbio.3000965.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb49/7723245/53184f06d87e/pbio.3000965.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb49/7723245/858ea2786fe4/pbio.3000965.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb49/7723245/59cf9999817c/pbio.3000965.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb49/7723245/a596eaa039f4/pbio.3000965.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb49/7723245/2707f3959e35/pbio.3000965.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb49/7723245/53184f06d87e/pbio.3000965.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb49/7723245/858ea2786fe4/pbio.3000965.g005.jpg

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