Tal Efrat, Shiloh Yosef
The David and Inez Myers Laboratory for Cancer Research, Department of Human Molecular Genetics and Biochemistry, Sackler School of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel.
Methods Mol Biol. 2017;1599:419-430. doi: 10.1007/978-1-4939-6955-5_30.
The ATM gene and its protein product, the ATM protein kinase, were identified as a result of attempts to understand the molecular basis of the genetic disorder, ataxia-telangiectasia (A-T). The cardinal symptom of A-T is neurodegeneration expressed primarily as progressive cerebellar atrophy. A major tool in the investigation of ATM functions in the cerebellum is cerebellar organotypic cultures, which allow cerebellar slices to live in culture for several weeks without losing their viability and organization. These cultures are amenable to various treatments and manipulations and provide a close look at Purkinje cells in their almost natural environment. We optimized the protocol for establishing and maintaining these cultures and provide here examples of readouts of the DNA damage response in cerebellar organotypic cultures treated with a DNA-damaging agent.
ATM基因及其蛋白质产物ATM蛋白激酶，是在试图了解遗传性疾病共济失调毛细血管扩张症（A-T）的分子基础的过程中被发现的。A-T的主要症状是神经退行性变，主要表现为进行性小脑萎缩。研究小脑ATM功能的一个主要工具是小脑器官型培养物，它能使小脑切片在培养中存活数周而不丧失其活力和组织结构。这些培养物适合进行各种处理和操作，并能让人们在几乎自然的环境中仔细观察浦肯野细胞。我们优化了建立和维持这些培养物的方案，并在此提供了用DNA损伤剂处理的小脑器官型培养物中DNA损伤反应读数的示例。
