Regional Cell-Specific Transcriptome Mapping Reveals Regulatory Complexity in the Adult Drosophila Midgut

Regional Cell-Specific Transcriptome Mapping Reveals Regulatory Complexity in the Adult Drosophila Midgut
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DOI:
10.1016/j.celrep.2015.06.009
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发表时间:
2015-07-14
期刊:
影响因子:
8.8
通讯作者:
Buchon, Nicolas
Buchon, Nicolas
中科院分区:
生物学1区
文献类型:
--
作者:
Dutta, Devanjali;Dobson, Adam J.;Buchon, Nicolas

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解读特定细胞类型对器官功能的贡献在实验上具有挑战性。果蝇中肠是一个动态的器官,具有五个形态和功能不同的区域(R1-R5),每个区域由多能肠干细胞(ISC)、肠母细胞(EB)、肠内分泌细胞(EE)、肠上皮细胞(EC)和内脏肌(VM)组成。为了表征该器官中的细胞特化和区域功能,我们生成了通过FACS从五个区域R1-R5中的每一个分离的所有五种细胞类型的RNA测序转录组。在这样做的过程中,我们确定细胞类型之间的转录差异,并记录每个细胞类型内的区域差异,定义进一步的专业化。我们验证了细胞特异性和区域Gal 4驱动程序;证明了转运蛋白Smvt和转录因子GATAe,Sna和Ptx 1在全球和区域ISC调控中的作用,并研究了感染后中肠细胞的转录反应。由此产生的转录组数据库(http:flygutseq.buchonlab.com)将促进区域化,稳态,免疫和细胞间相互作用的研究。
Deciphering contributions of specific cell types to organ function is experimentally challenging. The Drosophila midgut is a dynamic organ with five morphologically and functionally distinct regions (R1-R5), each composed of multipotent intestinal stem cells (ISCs), progenitor enteroblasts (EBs), enteroendocrine cells (EEs), enterocytes (ECs), and visceral muscle (VM). To characterize cellular specialization and regional function in this organ, we generated RNA-sequencing transcriptomes of all five cell types isolated by FACS from each of the five regions, R1-R5. In doing so, we identify transcriptional diversities among cell types and document regional differences within each cell type that define further specialization. We validate cell-specific and regional Gal4 drivers; demonstrate roles for transporter Smvt and transcription factors GATAe, Sna, and Ptx1 in global and regional ISC regulation, and study the transcriptional response of midgut cells upon infection. The resulting transcriptome database (http://flygutseq.buchonlab.com) will foster studies of regionalization, homeostasis, immunity, and cell-cell interactions.