Ascl1b and Neurod1, instead of Neurog3, control pancreatic endocrine cell fate in zebrafish

Lydie C Flasse,David G Stern,Bernard Peers,Virginie Von Berg,Justine L Pirson,Marianne L Voz,Isabelle Manfroid

doi:10.1186/1741-7007-11-78

Abstract

BackgroundNEUROG3 is a key regulator of pancreatic endocrine cell differentiation in mouse, essential for the generation of all mature hormone producing cells. It is repressed by Notch signaling that prevents pancreatic cell differentiation by maintaining precursors in an undifferentiated state.ResultsWe show that, in zebrafish, neurog3 is not expressed in the pancreas and null neurog3 mutant embryos do not display any apparent endocrine defects. The control of endocrine cell fate is instead fulfilled by two basic helix-loop-helix factors, Ascl1b and Neurod1, that are both repressed by Notch signaling. ascl1b is transiently expressed in the mid-trunk endoderm just after gastrulation and is required for the generation of the first pancreatic endocrine precursor cells. Neurod1 is expressed afterwards in the pancreatic anlagen and pursues the endocrine cell differentiation program initiated by Ascl1b. Their complementary role in endocrine differentiation of the dorsal bud is demonstrated by the loss of all hormone-secreting cells following their simultaneous inactivation. This defect is due to a blockage of the initiation of endocrine cell differentiation.ConclusionsThis study demonstrates that NEUROG3 is not the unique pancreatic endocrine cell fate determinant in vertebrates. A general survey of endocrine cell fate determinants in the whole digestive system among vertebrates indicates that they all belong to the ARP/ASCL family but not necessarily to the Neurog3 subfamily. The identity of the ARP/ASCL factor involved depends not only on the organ but also on the species. One could, therefore, consider differentiating stem cells into insulin-producing cells without the involvement of NEUROG3 but via another ARP/ASCL factor.

Highlights

NEUROG3 is a key regulator of pancreatic endocrine cell differentiation in mouse, essential for the generation of all mature hormone producing cells
Loss or dysfunction of endocrine insulin-secreting β-cells leads to diabetes, a widespread disease affecting more than 370 million people worldwide
We found that neurod1 and ascl1b expression is strongly increased in mib pancreas (Figure 4), indicating that ascl1b and neurod1 expression is repressed by Notch signaling, further suggesting a role of these two factors in endocrine differentiation

Summary

Introduction

NEUROG3 is a key regulator of pancreatic endocrine cell differentiation in mouse, essential for the generation of all mature hormone producing cells. Much of our knowledge on pancreas organogenesis relies on mouse genetic studies, the use of zebrafish has significantly contributed to the deciphering of mechanisms involved in the earliest phases of pancreas development [7,8,9,10,11,12] In this fish, the endoderm forms two converging sheets of cells by the end of gastrulation (10 hours post fertilization, hpf ). Appear the somatostatin-secreting δ-cells (17 hpf ), the ghrelin εcells (18 hpf ) and the glucagon-producing α-cells (21 hpf ) This first wave of endocrine cells is followed by a second wave coming from the ventral bud that forms from 32 hpf onwards [15,16,17]. While at 2 days post fertilization (dpf ), the vast majority of the endocrine cells is generated from the dorsal bud, at 12 days, a majority seems to derive from the ventral bud [16]

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