Department of Radiation Medicine, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, China.
Key Laboratory of Heavy Ion Radiation Biology and Medicine of Chinese Academy of Sciences, Institute of Modern Physics, Lanzhou, China.
J Cell Physiol. 2019 Dec;234(12):22439-22449. doi: 10.1002/jcp.28808. Epub 2019 May 13.
The mitochondrial proteins involved in spermatogenic cells apoptosis in zebrafish after carbon ion radiation (CIR) were screened. The relative biological effectiveness (RBE) of CIR in zebrafish testes was investigated. Apoptosis of testicular cells was measured within 24 hr following 1 and 4 Gy CIR. Immunoblotting was used to assess the levels of mitochondrial apoptotic proteins in testes, and proliferative and apoptotic spermatogenic cells were detected by immunofluorescence after CIR. Label-free quantitative (LFQ) and parallel reaction monitoring-based target proteomics (PRM) were combined to screen and validate differential mitochondrial proteins in testes between 4 Gy and control groups at 24 hr after CIR. The RBE of CIR in zebrafish testes was 1.48 ± 0.04, and induction of apoptosis by CIR was higher than that of X-rays in testicular cells. Mitochondrial apoptotic pathways play a crucial role in spermatogenic cells apoptosis after CIR, with 60 differential mitochondrial proteins identified. Among 20 target proteins, 12 were significantly upregulated, 2 were significantly downregulated in the 4 Gy CIR group. The results of PRM were consistent with label-free analysis. This is the first study to screen the differential mitochondrial proteins and provide useful information to understand the underlying mechanisms of spermatogenic cell apoptosis in zebrafish following CIR.
筛选了斑马鱼精子细胞凋亡后碳离子辐射（CIR）涉及的线粒体蛋白，并研究了 CIR 在斑马鱼睾丸中的相对生物效应（RBE）。在接受 1 和 4 Gy CIR 后 24 小时内测量睾丸细胞的凋亡情况。免疫印迹用于评估睾丸中线粒体凋亡蛋白的水平，并用 CIR 后免疫荧光检测增殖和凋亡的精原细胞。无标记定量（LFQ）和基于平行反应监测的靶向蛋白质组学（PRM）相结合，筛选和验证 CIR 后 24 小时 4 Gy 组和对照组睾丸中差异的线粒体蛋白。CIR 在斑马鱼睾丸中的 RBE 为 1.48±0.04，CIR 诱导的睾丸细胞凋亡高于 X 射线。线粒体凋亡途径在 CIR 后精子细胞凋亡中起关键作用，共鉴定出 60 种差异线粒体蛋白。在 20 个靶蛋白中，有 12 个明显上调，有 2 个明显下调。PRM 的结果与无标记分析一致。这是首次筛选差异线粒体蛋白的研究，为了解 CIR 后斑马鱼精子细胞凋亡的潜在机制提供了有用信息。
