Elucidating the mechanisms of chromosome length sensing by the synaptonemal complex

阐明联会复合体感知染色体长度的机制

• 批准号: 22K19272
• 负责人: Carlton Peter
• 金额: $ 4.16万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Research (Exploratory)
• 财政年份: 2022
• 资助国家: 日本
• 起止时间: 2022-06-30 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22K19272/
• 关键词: 減数分裂; 線虫; 染色体ダイナミクス; シナプトネマ複合体; in vivoイメージング; ライブイメージング

In this fiscal year we have improved our imaging techniques by making several adjustments to the in vivo conditions. We have succeeded in following the dynamic movement of mMaple3-labeled SYP-3 protein within and between meiotic chromosomes of C. elegans, while simultaneously visualizing the position of crossover designation sites by using eGFP-labeled COSA-1 protein. While both COSA-1 and non-activated mMaple3-SYP-3 are fluorescent in the green channel, we have found conditions where the COSA-1 foci that mark crossovers are unambiguously visible against the background of the linear signal characterizing SYP-3 staining. Thus, we were able to overcome the limitation of having only two fluorescent channels to measure three desired signals (non-activated mMaple3-SYP-3, activated mMaple3-SYP-3, and COSA-1). Further, we have developed an updated set of image analysis tools (https://github.com/carltonlab/carltonlab_imagej_tools) and simulation software (https://github.com/carltonlab/scsim) that we use to generate and test predictions of our diffusion and accumulation model.The graduate student leading this project has presented our work-in-progress at the Meiosis Gordon Conference in FY2022, in accordance with the research plan, and received valuable feedback from the research community.

在本财年，我们通过对体内条件进行了几次调整，改进了我们的成像技术。我们成功地跟踪了mMaple3标记的SYP-3蛋白在线虫减数分裂染色体内和减数分裂染色体之间的动态移动，同时利用EGFP标记的COSA-1蛋白显示了交叉指定位点的位置。虽然COSA-1和未激活的mMaple3-SYP-3在绿色通道中都是荧光的，但我们发现了标记交叉的COSA-1焦点在表征SYP-3染色的线性信号的背景下明确可见的情况。因此，我们能够克服仅有两个荧光通道来测量三个所需信号(非激活的mMaple3-SYP-3、激活的mMaple3-SYP-3和COSA-1)的限制。此外，我们已经开发了一套更新的图像分析工具(https://github.com/carltonlab/carltonlab_imagej_tools)和模拟软件(https://github.com/carltonlab/scsim))，我们使用它们来生成和测试我们的扩散和积累模型的预测。领导这个项目的研究生已经根据研究计划在2022财年的减数分裂戈登会议上展示了我们的工作进展情况，并从研究界收到了宝贵的反馈。

项目成果

Asymmetric partitioning of synaptonemal complex phosphorylation in fusion chromosomes of C. elegans

线虫融合染色体联会复合体磷酸化的不对称分配

• 发表时间: 2022
• 作者: RODRIGUEZ;Carlos
• 通讯作者: Carlos

Simulations of synaptonemal complex partitioning

联会复杂划分的模拟