Comprehensive single cell mRNA profiling reveals a detailed roadmap for pancreatic endocrinogenesis
Comprehensive single cell mRNA profiling reveals a detailed roadmap for pancreatic endocrinogenesis
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DOI：
10.1242/dev.173849
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发表时间：
2019-06-01
期刊：
影响因子：
4.6
通讯作者：
Bakhti, Mostafa
中科院分区：
文献类型：
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作者：
Bastidas-Ponce, Aimee;Tritschler, Sophie;Bakhti, Mostafa
破译内分泌细胞的诱导，规范和谱系分配在体内的机制将提供有价值的见解如何产生的胰岛。目前，内分泌祖细胞在发育过程中如何分离成不同的内分泌亚型尚不清楚。在这里，我们产生了一种新的神经元3（Ngn 3）-金星融合（NVF）报告小鼠系，密切反映了瞬时内源性Ngn 3蛋白表达。为了确定内分泌发生的体内路线图，我们对36，351个胰腺上皮细胞和NVF+细胞在次级转化期间进行了单细胞RNA测序。这使得Ngn 3（低）内分泌祖细胞，Ngn 3（高）内分泌前体，Fev（+）内分泌谱系和激素（+）内分泌亚型得以区分和时间分辨，并在逐步谱系限制步骤中描绘分子程序。引人注目的是，我们鉴定了58个新的标记基因，它们在7260个分析的Ngn 3表达细胞中显示出与Ngn 3相同的瞬时表达动力学。这些基因在内分泌前体细胞中的差异表达与其细胞命运分配到不同的内分泌细胞类型有关。因此，精确调节的NVF报告基因的产生使我们能够暂时解析内分泌谱系发育，以提供体内内分泌发生的细粒度单细胞分子谱。
Deciphering mechanisms of endocrine cell induction, specification and lineage allocation in vivo will provide valuable insights into how the islets of Langerhans are generated. Currently, it is ill defined how endocrine progenitors segregate into different endocrine subtypes during development. Here, we generated a novel neurogenin 3 (Ngn3)-Venus fusion (NVF) reporter mouse line, that closely mirrors the transient endogenous Ngn3 protein expression. To define an in vivo roadmap of endocrinogenesis, we performed single cell RNA sequencing of 36,351 pancreatic epithelial and NVF+ cells during secondary transition. This allowed Ngn3(low) endocrine progenitors, Ngn3(high) endocrine precursors, Fev(+) endocrine lineage and hormone(+) endocrine subtypes to be distinguished and time-resolved, and molecular programs during the step-wise lineage restriction steps to be delineated. Strikingly, we identified 58 novel signature genes that show the same transient expression dynamics as Ngn3 in the 7260 profiled Ngn3-expressing cells. The differential expression of these genes in endocrine precursors associated with their cell-fate allocation towards distinct endocrine cell types. Thus, the generation of an accurately regulated NVF reporter allowed us to temporally resolve endocrine lineage development to provide a fine-grained single cell molecular profile of endocrinogenesis in vivo.