使用 TUNEL 检测法研究果蝇 翅 盘细胞凋亡的实验方案。
Protocol to study cell death using TUNEL assay in imaginal discs.
机构信息
Department of Biology, University of Dayton, Dayton, OH 45469, USA.
Premedical Program, University of Dayton, Dayton, OH 45469, USA.
出版信息
STAR Protoc. 2022 Jan 27;3(1):101140. doi: 10.1016/j.xpro.2022.101140. eCollection 2022 Mar 18.
Cell death maintains tissue homeostasis by eliminating dispensable cells. Misregulation of cell death is seen in diseases like cancer, neurodegeneration, etc. Therefore, cell death assays like TUNEL have become reliable tools, where fragmented DNA of dying cells gets fluorescently labeled and can be detected under microscope. We used TUNEL assay in third-instar larval eye-antennal imaginal discs to label and quantify cell death. This assay is sensitive to detect DNA fragmentation, an important event, during apoptosis in retinal neurons. For complete details on the use and execution of this profile, please refer to Wang et al. (1999), Tare et al. (2011), and Mehta et al. (2021).
细胞死亡通过清除多余的细胞来维持组织内稳态。在癌症、神经退行性疾病等疾病中，细胞死亡的调节出现异常。因此，像 TUNEL 这样的细胞死亡检测方法已成为可靠的工具，凋亡过程中死亡细胞的 DNA 片段会被荧光标记，然后在显微镜下进行检测。我们在三龄幼虫的眼-触角imaginal 盘上使用 TUNEL 检测法对细胞死亡进行标记和定量。该检测法对视网膜神经元凋亡过程中的 DNA 片段化这一重要事件非常敏感。有关此方案的详细使用和执行信息，请参考 Wang 等人（1999 年）、Tare 等人（2011 年）和 Mehta 等人（2021 年）的研究。
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