@article{89a53db76d3b488a88a6d4e969fb2491,
title = "FUCCI tracking shows cell-cycle-dependent Neurog3 variation in pancreatic progenitors",
abstract = "During pancreas organogenesis, Neurog3HI endocrine-committing cells are generated from a population of Sox9+ mitotic progenitors with only a low level of Neurog3 transcriptional activity (Neurog3TA.LO). Low-level Neurog3 protein, in Neurog3TA.LO cells, is required to maintain their mitotic endocrine-lineage-primed status. Herein, we describe a Neurog3-driven FUCCI cell-cycle reporter (Neurog3P2A.FUCCI) derived from a Neurog3 BAC transgenic reporter that functions as a loxed cassette acceptor (LCA). In cycling Sox9+ Neurog3TA.LO progenitors, the majority of cells in S-G2-M phases have undetectable levels of Neurog3 with increased expression of endocrine progenitor markers, while those in G1 have low Neurog3 levels with increased expression of endocrine differentiation markers. These findings support a model in which variations in Neurog3 protein levels are coordinated with cell-cycle phase progression in Neurog3TA.LO progenitors with entrance into G1 triggering a concerted effort, beyond increasing Neurog3 levels, to maintain an endocrine-lineage-primed state by initiating expression of the downstream endocrine differentiation program prior to endocrine-commitment.",
keywords = "endocrine-biased, lineage priming, progenitor",
author = "Bechard, {Matthew E.} and Bankaitis, {Eric D.} and Alessandro Ustione and Piston, {David W.} and Magnuson, {Mark A.} and Wright, {Christopher V.E.}",
note = "Funding Information: We thank Atsushi Miyawaki (RIKEN Brain Science Institute) for the mKO2-hCdt1(30/120)/pCSII-EF-MCS and mVenus-hGem(1/110)/pCSII-EF-MCS plasmids. We also thank Dr. Douglas Mortlock (Vanderbilt) for his advice on the copy number estimation assay. This work utilized the Cell Imaging Shared Resource and Transgenic/ES Cell Shared Resource core facilities of the Vanderbilt Diabetes Research and Training Center funded by NIDDK grant 020593. Flow cytometry was performed in the VUMC Flow Cytometry Shared Resource supported by the Vanderbilt-Ingram Cancer Center (P30 CA68485) and the Vanderbilt Digestive Disease Research Center (DK0558404). Generation of Neurog3RG2 and Neurog3P2A.FUCCI mice was supported in part by the Beta Cell Biology Consortium Mouse ES Cell Core funded by the NIDDK (U01DK072473). We thank Anna Means, Guoqiang Gu, and members of the Wright/Gu labs for discussions. This study was supported by the NIH/NIDDK (U01DK089570) and an American Heart Association fellowship to MB (13POST14240011). Funding Information: Cell Imaging Shared Resource and Transgenic/ES Cell Shared Resource core facilities of the Vanderbilt Diabetes Research and Training Center funded by NIDDK, Grant Number: 020593, Vanderbilt-Ingram Cancer Center, Grant Number: P30 CA68485, Vanderbilt Diges tive Disease Research Center, Grant Num ber: DK0558404, Beta Cell Biology Consortium Mouse ES Cell Core funded by the NIDDK, Grant Number: U01DK072473, NIH/NIDDK, Grant Num ber: U01DK089570, American Heart Asso ciation fellowship to MB, Grant Number: 13POST14240011. Publisher Copyright: {\textcopyright} 2017 Wiley Periodicals, Inc.",
year = "2017",
month = sep,
doi = "10.1002/dvg.23050",
language = "English",
volume = "55",
journal = "Genesis",
issn = "1526-954X",
number = "9",
}