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.

在这个财政年度，我们通过对体内条件进行了几次调整，改善了成像技术。我们成功地遵循了秀丽隐杆线虫的减数分裂染色体内和之间的Mmaple3标记的SYP-3蛋白的动态运动，同时使用EGFP标记的COSA-COSA-1蛋白同时可视化交叉名称位点的位置。尽管COSA-1和未激活的MMAPLE3-SYP-3都是绿色通道中的荧光，但我们发现了在表征SYP-3染色的线性信号的背景下明确可见的COSA-1灶。因此，我们能够克服只有两个荧光通道来测量三个所需信号（未激活的MMAPLE3-SYP-3，激活的MMAPLE3-SYP-3和COSA-1）的局限性。此外，我们已经开发了一组更新的图像分析工具（https://github.com/carltonlab/carltonlab/carltonlab_imagej_tools）和仿真软件（https：//github.com/carltonlab/scsim），我们用来为我们的扩散和累积的模型来启示和测试我们的累积模型。根据研究计划，在2022财年举行的戈登会议，并收到了研究界的宝贵反馈。

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