Imaging functional neural circuits in zebrafish with a new GCaMP and the Gal4FF-UAS system.
Imaging functional neural circuits in zebrafish with a new GCaMP and the Gal4FF-UAS system.
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DOI：
10.4161/cib.4.5.15848
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发表时间：
2011-09-01
影响因子：
--
通讯作者：
Kawakami, Koichi
中科院分区：
文献类型：
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作者：
Muto, Akira;Kawakami, Koichi
测量神经元集合的活动是了解神经元网络如何组织和运作的重要步骤。神经元的电兴奋通过电压门控钙离子通道引起钙内流，这可以通过使用荧光钙探针的钙成像来监测。DNA编码的钙指示剂（DECIs），如cameleon和GCaMP已被开发用于特异性标记神经元亚群。然而，在许多情况下，已经在体外开发和测试的DECIs并不总是在体内显示出预期的性能。有必要提高其灵敏度，并根据生理条件调整其动态范围。在我们最近的研究中，我们开发了GCaMP的改进版本，并使用转基因斑马鱼在体内测试其性能。通过Gal 4FF-UAS系统将新的GCaMP与靶向基因表达相结合，我们成功地成像了斑马鱼幼虫自发收缩期间脊髓运动回路的活动。此外，我们在这里报告庚醇，间隙连接阻滞剂，可以改变运动回路的时空激活模式。因此，我们证明，钙成像与GCaMP是强大的分析神经元的活动在正常和非扰动条件下。
Measurement of the activity of neuronal ensembles is an essential step to understand how the neuronal network is organized and functioning. Electrical excitation of neurons causes calcium influx via voltage-gated calcium ion channels, which can be monitored by calcium imaging using fluorescent calcium probes. DNA-encoded calcium indicators (DECIs) such as cameleon and GCaMP have been developed to specifically label a subpopulation of neurons. However, in many cases, DECIs that had been developed and tested in vitro did not always show expected performance in vivo. It is necessary to increase its sensitivity and also to adjust its dynamic range to the physiological conditions. In our recent study, we developed an improved version of GCaMP and tested its performance in vivo using transgenic zebrafish. By combining the new GCaMP with targeted gene expression via the Gal4FF-UAS system, we successfully imaged the activity of the spinal motor circuit during spontaneous contractions of zebrafish larvae. Further we report here that heptanol, a gap junction blocker, could alter the spatiotemporal activation pattern of the motor circuit. Thus, we demonstrate that calcium imaging with GCaMP is powerful to analyze neuronal activities under normal and pharmacologically perturbed conditions.