Beaudier Pierre, Devès Guillaume, Plawinski Laurent, Dupuy Denis, Barberet Philippe, Seznec Hervé
University Bordeaux, CNRS, LP2I, UMR 5797, 33170 Gradignan, France.
University Bordeaux, INSERM, U1212, 33607 Pessac, France.
Biology (Basel). 2023 Oct 27;12(11):1372. doi: 10.3390/biology12111372.
We describe a methodology to manipulate () and irradiate the stem progenitor gonad region using three MeV protons at a specific developmental stage (L1). The consequences of the targeted irradiation were first investigated by considering the organogenesis of the vulva and gonad, two well-defined and characterized developmental systems in . In addition, we adapted high-throughput analysis protocols, using cell-sorting assays (COPAS) and whole transcriptome analysis, to the limited number of worms (>300) imposed by the selective irradiation approach. Here, the presented status report validated protocols to (i) deliver a controlled dose in specific regions of the worms; (ii) immobilize synchronized worm populations (>300); (iii) specifically target dedicated cells; (iv) study the radiation-induced developmental alterations and gene induction involved in cellular stress (heat shock protein) and cuticle injury responses that were found.
我们描述了一种方法,可在特定发育阶段(L1)使用3兆电子伏特质子对()进行操作并照射干细胞祖性腺区域。通过考虑外阴和性腺的器官发生,这是两种在中定义明确且特征显著的发育系统,首先研究了靶向照射的后果。此外,我们针对选择性照射方法所限制的有限数量的蠕虫(>300条),采用细胞分选测定(COPAS)和全转录组分析等高通量分析方案。在此,所呈现的状态报告验证了以下方案:(i)在蠕虫的特定区域递送可控剂量;(ii)固定同步化的蠕虫群体(>300条);(iii)特异性靶向特定细胞;(iv)研究辐射诱导的发育改变以及所发现的涉及细胞应激(热休克蛋白)和表皮损伤反应的基因诱导。