Abstract
Basic helix-loop-helix genes, particularly proneural genes, are well-described triggers of cell differentiation, yet information on their dynamics is limited, notably in human development. Here, we focus on Neurogenin 3 (NEUROG3), which is crucial for pancreatic endocrine lineage initiation. By monitoring both NEUROG3 gene expression and protein in single cells using a knockin dual reporter in 2D and 3D models of human pancreas development, we show an approximately 2-fold slower expression of human NEUROG3 than that of the mouse. We observe heterogeneous peak levels of NEUROG3 expression and reveal through long-term live imaging that both low and high NEUROG3 peak levels can trigger differentiation into hormone-expressing cells. Based on fluorescence intensity, we statistically integrate single-cell transcriptome with dynamic behaviors of live cells and propose a data-mapping methodology applicable to other contexts. Using this methodology, we identify a role for KLK12 in motility at the onset of NEUROG3 expression.
| Originalsprog | Engelsk |
|---|---|
| Tidsskrift | Developmental Cell |
| Vol/bind | 58 |
| Udgave nummer | 21 |
| Sider (fra-til) | 2292-2308.e6 |
| Antal sider | 24 |
| ISSN | 1534-5807 |
| DOI | |
| Status | Udgivet - 2023 |
Bibliografisk note
Funding Information:The authors would like to thank DanStem Platforms of Genomics, Stem Cell Culture, Flow Cytometry (especially Paul van Dieken), and Imaging for their training, technical expertise, support, and the use of instruments. Additionally, we thank the Scientific Computing Facility (especially Noreen Walker, Gayathri Nadar, and Andre Gohr), Organoid and Stem Cell Facility, FACS Facility, and Light Microscopy Facility at MPI-CBG for their support and technical help. The Deep Sequencing Facility of the CMCB Technology Platform (Center for Regenerative Therapy in Dresden) provided invaluable service with library preparation and sequencing for the RNA sequencing experiments. Special thanks to Jifeng Liu and Allison Lewis for their support. The Novo Nordisk Foundation Center for Stem Cell Biology was supported by a Novo Nordisk Foundation grant number NNF17CC0027852 . B.S.B.-T. is supported by the Copenhagen Bioscience PhD program financed by the Novo Nordisk Fonden ( NNF16CC0020994 ).
Funding Information:
The authors would like to thank DanStem Platforms of Genomics, Stem Cell Culture, Flow Cytometry (especially Paul van Dieken), and Imaging for their training, technical expertise, support, and the use of instruments. Additionally, we thank the Scientific Computing Facility (especially Noreen Walker, Gayathri Nadar, and Andre Gohr), Organoid and Stem Cell Facility, FACS Facility, and Light Microscopy Facility at MPI-CBG for their support and technical help. The Deep Sequencing Facility of the CMCB Technology Platform (Center for Regenerative Therapy in Dresden) provided invaluable service with library preparation and sequencing for the RNA sequencing experiments. Special thanks to Jifeng Liu and Allison Lewis for their support. The Novo Nordisk Foundation Center for Stem Cell Biology was supported by a Novo Nordisk Foundation grant number NNF17CC0027852. B.S.B.-T. is supported by the Copenhagen Bioscience PhD program financed by the Novo Nordisk Fonden (NNF16CC0020994). Conceptualization, B.S.B.-T. Y.H.K. and A.G.-B.; methodology, B.S.B.-T. Y.H.K. L.H. B.H.L. F.L. C.Z. and A.G.-B.; investigation, B.S.B.-T. and J.V.D.; resources, A.G.-B.; writing – original draft, B.S.B.-T. Y.H.K. and A.G.-B.; writing – review & editing, B.S.B.-T. L.H. F.L. C.Z. Y.H.K. and A.G.-B.; visualization, B.S.B.-T.; supervision, Y.H.K. and A.G.-B. The authors declare no competing interests.
Publisher Copyright:
© 2023 The Author(s)