Department of Biomedical Sciences, Florida State University, College of Medicine, Tallahassee, Florida, United States of America.
Program of Neuroscience, Florida State University, Tallahassee, Florida, United States of America.
PLoS Genet. 2021 Apr 26;17(4):e1009240. doi: 10.1371/journal.pgen.1009240. eCollection 2021 Apr.
Examining the role of chromatin modifications and gene expression in neurons is critical for understanding how the potential for behaviors are established and maintained. We investigate this question by examining Drosophila melanogaster fru P1 neurons that underlie reproductive behaviors in both sexes. We developed a method to purify cell-type-specific chromatin (Chromatag), using a tagged histone H2B variant that is expressed using the versatile Gal4/UAS gene expression system. Here, we use Chromatag to evaluate five chromatin modifications, at three life stages in both sexes. We find substantial changes in chromatin modification profiles across development and fewer differences between males and females. Additionally, we find chromatin modifications that persist in different sets of genes from pupal to adult stages, which may point to genes important for cell fate determination in fru P1 neurons. We generated cell-type-specific RNA-seq data sets, using translating ribosome affinity purification (TRAP). We identify actively translated genes in fru P1 neurons, revealing novel stage- and sex-differences in gene expression. We also find chromatin modification enrichment patterns that are associated with gene expression. Next, we use the chromatin modification data to identify cell-type-specific super-enhancer-containing genes. We show that genes with super-enhancers in fru P1 neurons differ across development and between the sexes. We validated that a set of genes are expressed in fru P1 neurons, which were chosen based on having a super-enhancer and TRAP-enriched expression in fru P1 neurons.
研究染色质修饰和基因表达在神经元中的作用对于理解行为潜力是如何建立和维持的至关重要。我们通过研究果蝇 melanogaster fru P1 神经元来探究这个问题,这些神经元是两性生殖行为的基础。我们开发了一种使用标记的组蛋白 H2B 变体来纯化细胞类型特异性染色质(Chromatag)的方法,该变体使用多功能 Gal4/UAS 基因表达系统表达。在这里,我们使用 Chromatag 在两性的三个生命阶段评估了五种染色质修饰。我们发现染色质修饰谱在发育过程中发生了很大变化,而雄性和雌性之间的差异较小。此外,我们发现了在蛹至成虫阶段在不同基因集中持续存在的染色质修饰,这可能指向 fru P1 神经元中对细胞命运决定很重要的基因。我们使用翻译核糖体亲和纯化(TRAP)生成了细胞类型特异性的 RNA-seq 数据集。我们鉴定了 fru P1 神经元中活跃翻译的基因,揭示了在基因表达方面的新的阶段和性别差异。我们还发现了与基因表达相关的染色质修饰富集模式。接下来,我们使用染色质修饰数据来鉴定细胞类型特异性的超级增强子包含基因。我们表明,在 fru P1 神经元中具有超级增强子的基因在发育过程中和两性之间存在差异。我们验证了一组基因在 fru P1 神经元中表达,这些基因是根据在 fru P1 神经元中具有超级增强子和 TRAP 富集表达而选择的。