连续光照转暗培养联合药物处理实现衣藻细胞的高水平同步化

单细胞衣藻(Chlamydomonas)是光合作用和植物细胞周期等生物学过程研究的一个重要模式系统, 同步化培养是进行相关研究的必要手段。该研究探索了连续光照转暗培养联合细胞周期阻断剂实现莱茵衣藻(Chlamydomonas reinhardtii)细胞高水平同步化的新方法, 并利用流式细胞术对同步化程度进行了精确的分析。结果表明, 连续光照转暗培养或联合S期阻断剂可以使衣藻细胞同步化到G1期或G1/S期边界; 连续光照转暗培养联合M期阻断剂或者在“加入-释放”S期阻断剂后再加入M期阻断剂可以使衣藻细胞同步化到M期, 同步化水平可达80%。具体的同步化培养步骤要根据研究对象(特别是某些衣藻突变株系)的特性和研究目的确定。

Highly Synchronized Chlamydomonas reinhardtii Cells by Dark-shifting a Continuous-light Culture Combined with Drugs

Chlamydomonas reinhardtii, a unicellular eukaryotic green alga, has been a powerful model system for understandingseveral fundamental biological processes such as photosynthesis and plant cell-cycle regulation. An artificialcell-cycle synchronization system is necessary for these studies. Here, we investigated a new method for synchronizingChlamydomonas cell-wall-less cells, with which a high level of synchrony can be achieved. The degree of synchrony wasanalysed by flow cytometry. The cells are grown asynchronously in continuous light, then grown for 24 h in the dark(dark-shifting a continuous-light culture). At the end of the dark period, most cells (>80%) are arrested in the G1 phase. Ifnecessary, an S-phase blocking agent can be used to treat the dark-shifted cells. When the dark-shifted cells are treatedwith M-phase blocking agent alone or with the S-phase and M-phase blocking agent sequentially, a cell population withmore than 80% of cells at the M phase can be achieved. A specific synchronous procedure needs to be selected accordingto the properties of a certain Chlamydomonas strain (especially certain mutants) and the requirement of research.