一种用于树突退变和修复的光开关检测系统。

A photo-switchable assay system for dendrite degeneration and repair in .

机构信息

Department of Physiology, University of California, San Francisco, CA 94143.

Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143.

出版信息

Proc Natl Acad Sci U S A. 2022 Aug 23;119(34):e2204577119. doi: 10.1073/pnas.2204577119. Epub 2022 Aug 15.

Abstract

Neurodegeneration arising from aging, injury, or diseases has devastating health consequences. Whereas neuronal survival and axon degeneration have been studied extensively, much less is known about how neurodegeneration affects dendrites, in part due to the limited assay systems available. To develop an assay for dendrite degeneration and repair, we used photo-switchable caspase-3 (caspase-Light-Oxygen-Voltage-sensing [caspase-LOV]) in peripheral class 4 dendrite arborization (c4da) neurons to induce graded neurodegeneration by adjusting illumination duration during development and adulthood in . We found that both developing and mature c4da neurons were able to survive while sustaining mild neurodegeneration induced by moderate caspase-LOV activation. Further, we observed active dendrite addition and dendrite regeneration in developing and mature c4da neurons, respectively. Using this assay, we found that the mouse Wallerian degeneration slow (Wld) protein can protect c4da neurons from caspase-LOV-induced dendrite degeneration and cell death. Furthermore, our data show that Wld can reduce dendrite elimination without affecting dendrite addition. In summary, we successfully established a photo-switchable assay system in both developing and mature neurons and used Wld as a test case to study the mechanisms underlying dendrite regeneration and repair.

摘要

神经退行性变源于衰老、损伤或疾病,会对健康造成严重后果。尽管神经元存活和轴突退化已得到广泛研究,但对于神经退行性变如何影响树突,人们知之甚少,部分原因是可用的检测系统有限。为了开发一种用于树突退化和修复的检测方法,我们在周围类 4 树突分支(c4da)神经元中使用光可切换的半胱天冬酶-3(caspase-Light-Oxygen-Voltage-sensing [caspase-LOV]),通过在发育和成年期间调整光照持续时间来诱导分级神经退行性变。我们发现,发育中和成熟的 c4da 神经元在维持适度 caspase-LOV 激活引起的轻度神经退行性变的同时能够存活。此外,我们观察到发育中和成熟的 c4da 神经元中分别有活跃的树突添加和树突再生。使用该检测方法,我们发现小鼠 Wallerian 变性慢(Wld)蛋白可以保护 c4da 神经元免受 caspase-LOV 诱导的树突退化和细胞死亡。此外,我们的数据表明,Wld 可以减少树突消除而不影响树突添加。总之,我们成功地在发育中和成熟神经元中建立了光可切换的检测系统,并将 Wld 作为一个测试案例来研究树突再生和修复的机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/159a/9407391/5ecb73fd80ac/pnas.2204577119fig01.jpg

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