Comparison of GCaMP3 and GCaMP6f for studying astrocyte Ca2+ dynamics in the awake mouse brain.
Comparison of GCaMP3 and GCaMP6f for studying astrocyte Ca2+ dynamics in the awake mouse brain.
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DOI：
10.1371/journal.pone.0181113
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Paukert M
中科院分区：
文献类型：
--
作者：
Ye L;Haroon MA;Salinas A;Paukert M
近年来，越来越清楚的是，星形胶质细胞在神经过程中发挥的作用比传统观点认为的支持细胞要积极得多。虽然不是电兴奋，星形胶质细胞表现出不同的Ca2+动态跨空间和时间尺度，或多或少依赖于动物的行为状态。Ca2+动力学范围从全球海拔持续数秒，包括索马最精细的过程，短海拔仅限于所谓的微区内精细的过程。星形胶质细胞Ca2+动力学的研究特别受益于遗传编码钙指标（GECIs）的发展。GECI表达可以以细胞类型特异性方式非侵入性地实现，并且它可以遗传靶向亚细胞结构域。GCaMP家族是一组来源于绿色荧光蛋白的GECIs，在过去十年中经历了一些最快的进展。因此，我们现在面临的挑战是，需要比较不同版本的GECIs获得的已发表数据。为了提供一些指导，我们在这里比较了表达成熟的GCaMP3或日益流行的GCaMP6f（特别是在星形胶质细胞中）的清醒转基因小鼠中不同尺度的Ca 2+动态。我们发现，运动诱导的全球钙离子升高皮质星形胶质细胞显示只有轻微的动力学差异和他们的表观动态范围内的钙离子传感没有什么不同。相比之下，Ca2+波的过程和微域Ca2+瞬变更容易检测GCaMP6f。我们的研究结果表明，行为状态依赖性的全球星形胶质细胞钙离子反应可以与GCaMP3或GCaMP6f，而后者更适合于空间限制的弱和快的钙离子动力学的研究。
In recent years it has become increasingly clear that astrocytes play a much more active role in neural processes than the traditional view of them as supporting cells suggests. Although not electrically excitable, astrocytes exhibit diverse Ca2+ dynamics across spatial and temporal scales, more or less dependent on the animal's behavioral state. Ca2+ dynamics range from global elevations lasting multiple seconds encompassing the soma up to the finest processes, to short elevations restricted to so-called microdomains within fine processes. Investigations of astrocyte Ca2+ dynamics have particularly benefitted from the development of Genetically-Encoded Calcium Indicators (GECIs). GECI expression can be achieved non-invasively in a cell type-specific manner and it can be genetically targeted to subcellular domains. The GCaMP family, a group of GECIs derived from the green fluorescent protein, has experienced some of the fastest advancements during the past decade. As a consequence we are now facing the challenge of needing to compare published data obtained with different versions of GECIs. With the intention to provide some guidance, here we compared Ca2+ dynamics across scales in awake transgenic mice expressing either the well-established GCaMP3, or the increasingly popular GCaMP6f, specifically in astrocytes. We found that locomotion-induced global Ca2+ elevations in cortical astrocytes displayed only minor kinetic differences and their apparent dynamic ranges for Ca2+ sensing were not different. In contrast, Ca2+ waves in processes and microdomain Ca2+ transients were much more readily detectable with GCaMP6f. Our findings suggest that behavioral state-dependent global astrocyte Ca2+ responses can be studied with either GCaMP3 or GCaMP6f whereas the latter is more appropriate for studies of spatially restricted weak and fast Ca2+ dynamics.
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