Autophagy Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India.
Chronobiology and Behavioural Neurogenetics Laboratory, Neuroscience Unit, Bangalore, India.
Methods Mol Biol. 2024;2761:97-120. doi: 10.1007/978-1-0716-3662-6_9.
Neuronal synapse dysfunction is a key characteristic of several neurodegenerative disorders, such as Alzheimer's disease, spinocerebellar ataxias, and Huntington's disease. Modeling these disorders to study synaptic dysfunction requires a robust and reproducible method for assaying the subtle changes associated with synaptopathies in terms of structure and function of the synapses. Drosophila melanogaster neuromuscular junctions (NMJs) serve as good models to study such alterations. Further, modifications in the microenvironment of synapses can sometimes reflect in the behavior of the animal, which can also be assayed in a high-throughput manner. The methods outlined in this chapter highlight assays to study the behavioral changes associated with synaptic dysfunction in a spinocerebellar ataxia type 3 (SCA3) model. Further, molecular assessment of alterations in NMJ structure and function is also summarized, followed by effects of autophagy pathway upregulation in providing neuroprotection. These methods can be further extended and modified to study the therapeutic effects of drugs or small molecules in providing neuroprotection for any synaptopathy models.
神经元突触功能障碍是几种神经退行性疾病的一个关键特征,如阿尔茨海默病、脊髓小脑共济失调和亨廷顿病。为了研究突触功能障碍,需要一种稳健且可重复的方法来检测与突触病相关的细微变化,包括突触的结构和功能。果蝇肌肉突触(NMJs)可作为研究这些变化的良好模型。此外,突触微环境的改变有时会反映在动物的行为上,这也可以以高通量的方式进行检测。本章中概述的方法突出了用于研究脊髓小脑共济失调 3 型 (SCA3) 模型中与突触功能障碍相关的行为变化的检测方法。此外,还总结了 NMJ 结构和功能改变的分子评估,以及自噬途径上调在提供神经保护方面的作用。这些方法可以进一步扩展和修改,以研究药物或小分子在为任何突触病模型提供神经保护方面的治疗效果。