Measurement of autophagy flux in the nervous system in vivo.
Measurement of autophagy flux in the nervous system in vivo.
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DOI：
10.1038/cddis.2013.421
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发表时间：
2013-11-14
影响因子：
9
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中科院分区：
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准确的方法来测量体内神经元中的自噬活性是不可用的，并且大多数研究是基于自噬标记物的相关和静态测量，导致相互矛盾的解释。自噬是一种重要的稳态过程，参与细胞器和蛋白质聚集体等多种细胞成分的降解。自噬损伤正在成为许多疾病中驱动神经变性的相关因素。此外，调节自噬的策略已被证明可以提供针对神经变性的保护。在这里，我们描述了一种新的和简单的策略，表达自噬流量报告在成年动物的神经系统的脑室内交付的腺相关病毒（AAV）到新生小鼠。使用这种方法，我们在外周和中枢神经系统的神经元中有效地表达了单体串联mCherry-GFP-LC 3构建体，允许在药理学和疾病环境中测量自噬活性。
Accurate methods to measure autophagic activity in vivo in neurons are not available, and most of the studies are based on correlative and static measurements of autophagy markers, leading to conflicting interpretations. Autophagy is an essential homeostatic process involved in the degradation of diverse cellular components including organelles and protein aggregates. Autophagy impairment is emerging as a relevant factor driving neurodegeneration in many diseases. Moreover, strategies to modulate autophagy have been shown to provide protection against neurodegeneration. Here we describe a novel and simple strategy to express an autophagy flux reporter in the nervous system of adult animals by the intraventricular delivery of adeno-associated viruses (AAV) into newborn mice. Using this approach we efficiently expressed a monomeric tandem mCherry-GFP-LC3 construct in neurons of the peripheral and central nervous system, allowing the measurement of autophagy activity in pharmacological and disease settings.