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Development. 2019 Jun 17;146(12). pii: dev173849. doi: 10.1242/dev.173849.

Comprehensive single cell mRNA profiling reveals a detailed roadmap for pancreatic endocrinogenesis.

Bastidas-Ponce A1,2,3,4, Tritschler S1,5,6, Dony L5,6,7, Scheibner K1,2,3,4, Tarquis-Medina M1,2,3,4, Salinno C1,2,3,4, Schirge S1,2,3, Burtscher I1,2,3, Böttcher A1,2,3, Theis FJ8,9, Lickert H10,2,3,4, Bakhti M10,2,3.

Author information

1
Institute of Diabetes and Regeneration Research, Helmholtz Zentrum München, D-85764 Neuherberg, Germany.
2
German Center for Diabetes Research (DZD), D-85764 Neuherberg, Germany.
3
Institute of Stem Cell Research, Helmholtz Zentrum München, D-85764 Neuherberg, Germany.
4
Technical University of Munich, School of Medicine, 81675 Munich, Germany.
5
Institute of Computational Biology, Helmholtz Zentrum München, D-85764 Neuherberg, Germany.
6
Technical University of Munich, School of Life Sciences Weihenstephan, 85354 Freising, Germany.
7
Max Planck Institute of Psychiatry, Kraepelinstr. 2-10, 80804 Munich, Germany.
8
Institute of Computational Biology, Helmholtz Zentrum München, D-85764 Neuherberg, Germany fabian.theis@helmholtz-muenchen.de heiko.lickert@helmholtz-muenchen.de mostafa.bakhti@helmholtz-muenchen.de.
9
Technical University of Munich, Department of Mathematics, 85748 Garching b. Munich, Germany.
10
Institute of Diabetes and Regeneration Research, Helmholtz Zentrum München, D-85764 Neuherberg, Germany fabian.theis@helmholtz-muenchen.de heiko.lickert@helmholtz-muenchen.de mostafa.bakhti@helmholtz-muenchen.de.

Abstract

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.

KEYWORDS:

Endocrine cell allocation; Endocrine progenitor-precursor; Endocrinogenesis; Mouse; Neurog3; Single cell RNA sequencing

PMID:
31160421
DOI:
10.1242/dev.173849

Conflict of interest statement

Competing interestsThe authors declare no competing or financial interests.

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