Characterisation of TET function during indirect flight muscle development

Enzymes of the Ten Eleven Translocation (TET) family play a key role in the regulation of gene expression in many species by oxidizing 5-methylcytosine (5mC), a prominent epigenetic mark, into 5-hydroxymethylcytosine (5hmC). Yet, TET proteins also have non-canonical modes of action beyond 5mC oxidation, notably in Drosophila, whose genomes is devoid of 5mC. Here, we assessed TET function during the development of the adult indirect flight muscles which are derived from a population of progenitors located on the larval wing imaginal discs. Overall design: All experiments were performed with biological replicates using dissected third instar larval wing discs or dissected pupal indirect flight muscles. w1118 (wt) or TET-GFP knock-in flies were used as control lines. Drosophila homozygous for Tet null or Tet catalytic dead alleles were used as test lines.

TET家族（Ten Eleven Translocation）的酶，通过将一类重要的表观遗传标记——5-甲基胞嘧啶（5-methylcytosine, 5mC）氧化为5-羟甲基胞嘧啶（5-hydroxymethylcytosine, 5hmC），在诸多物种的基因表达调控中发挥关键作用。然而，TET蛋白还存在超越5mC氧化的非经典作用模式，这一点在基因组中不含5mC的果蝇（Drosophila）中尤为显著。本研究针对由幼虫翅成虫盘上的祖细胞群体分化而来的成体间接飞行肌发育过程中的TET功能进行了评估。

实验设计：所有实验均采用生物学重复，实验材料为解剖获取的三龄幼虫翅成虫盘或蛹期间接飞行肌。本研究以w1118（野生型，wt）或TET-GFP敲入果蝇作为对照品系，以携带Tet纯合缺失等位基因或Tet催化失活等位基因的纯合果蝇作为实验组品系。