Long-term optical brain imaging in live adult fruit flies.

Long-term optical brain imaging in live adult fruit flies.
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DOI:
10.1038/s41467-018-02873-1
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发表时间:
2018-02-28
影响因子:
16.6
通讯作者:
Schnitzer MJ
Schnitzer MJ
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Huang C;Maxey JR;Sinha S;Savall J;Gong Y;Schnitzer MJ

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细胞形态学和生理学的活体延时显微镜研究对于理解脑功能至关重要,但在果蝇这种关键的模式物种中是不可行的。在这里,我们使用激光显微手术来创建一个慢性苍蝇准备,用于长达50天的神经结构和动力学重复成像。在蝇菇体神经元中,我们对轴突钮扣进行了10天的跟踪,并在7周内记录了气味诱发的钙瞬变。此外,通过电压成像来解析个体动作电位,我们监测了机械应力下果蝇多巴胺神经元的尖峰可塑性。应激24 h后,PPL1-α ' 3多巴胺神经元的尖峰率升高,但PPL1-α ' 2α2多巴胺神经元的尖峰率没有升高。总的来说,我们的慢性制备与广泛的光学技术兼容,并且能够对许多以前无法在活蝇中解决的生物学问题进行纵向研究。到目前为止,对苍蝇大脑进行一天以上的延时成像研究是不可能的。在这里,作者提出了一种激光显微手术方法,在苍蝇角皮上创建一个永久窗口,使神经结构和动力学的延时成像长达10-50天。
Time-lapse in vivo microscopy studies of cellular morphology and physiology are crucial toward understanding brain function but have been infeasible in the fruit fly, a key model species. Here we use laser microsurgery to create a chronic fly preparation for repeated imaging of neural architecture and dynamics for up to 50 days. In fly mushroom body neurons, we track axonal boutons for 10 days and record odor-evoked calcium transients over 7 weeks. Further, by using voltage imaging to resolve individual action potentials, we monitor spiking plasticity in dopamine neurons of flies undergoing mechanical stress. After 24 h of stress, PPL1-α’3 but not PPL1-α’2α2 dopamine neurons have elevated spike rates. Overall, our chronic preparation is compatible with a broad range of optical techniques and enables longitudinal studies of many biological questions that could not be addressed before in live flies. Time-lapse imaging studies of more than a day in the fly brain have been infeasible until now. Here the authors present a laser microsurgery approach to create a permanent window in the fly cuticle to enable time-lapse imaging of neural architecture and dynamics for up to 10–50 days.
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