Recovery of Tetrahymena thermophila from degranulation. Recovery of Tetrahymena thermophila from degranulation

The object was to identify genes that are upregulated during the recovery of cells from stimulation of granule exocytosis, when new granules are being synthesized. The granules are also known as mucocysts. To distinguish between genes upregulated due to demands of granule synthesis, vs genes upregulated in response to the exocytic stimulus per se, we exploited a mutant cell line, MN173, that does not release its granules on stimulation. Overall design: Total RNA was isolated from cells prior to stimulation of exocytosis, and then 60 minutes post stimulation. In parallel, cells from the MN173 mutant line were treated and processed equivalently.

本研究旨在鉴定细胞经颗粒胞吐刺激后的恢复阶段中上调的基因，此时细胞正合成新的颗粒。此类颗粒亦被称为粘液囊泡（mucocysts）。为区分两类上调基因：一类是因颗粒合成需求而上调的基因，另一类是直接响应胞吐刺激本身而上调的基因，我们采用了一株无法在刺激条件下释放颗粒的突变细胞系MN173。实验整体设计：分别于胞吐刺激前、刺激后60分钟提取细胞总RNA；同时，对MN173突变细胞系开展完全一致的处理与样本制备流程。