Cdon mutation and fetal alcohol converge on Nodal signaling in a mouse model of holoprosencephaly.

Mingi Hong,Robert S Krauss,Anna Christa,Annabel Christ,Thomas E Willnow

doi:10.7554/elife.60351

Mingi Hong, Robert S Krauss + Show 3 more

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https://doi.org/10.7554/elife.60351

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Abstract

Holoprosencephaly (HPE), a defect in midline patterning of the forebrain and midface, arises ~1 in 250 conceptions. It is associated with predisposing mutations in the Nodal and Hedgehog (HH) pathways, with penetrance and expressivity graded by genetic and environmental modifiers, via poorly understood mechanisms. CDON is a multifunctional co-receptor, including for the HH pathway. In mice, Cdon mutation synergizes with fetal alcohol exposure, producing HPE phenotypes closely resembling those seen in humans. We report here that, unexpectedly, Nodal signaling is a major point of synergistic interaction between Cdon mutation and fetal alcohol. Window-of-sensitivity, genetic, and in vitro findings are consistent with a model whereby brief exposure of Cdon mutant embryos to ethanol during gastrulation transiently and partially inhibits Nodal pathway activity, with consequent effects on midline patterning. These results illuminate mechanisms of gene-environment interaction in a multifactorial model of a common birth defect.

Highlights

Many common structural birth defects appear to arise from a complex and ill-defined combination of genetic and environmental factors (Krauss and Hong, 2016)
We demonstrate that the window of sensitivity to EtOH-induced HPE in 129S6 Cdon-/- mice is closed by embryonic day (E) 7.5, with the sensitivity period overlapping Nodal-mediated specification of the PCP from the anterior primitive streak (APS)
Expression of Lefty2, a direct Nodal pathway target gene, was reduced at E7.25 in EtOH-treated Cdon-/- embryos (Figure 2). These results showed that the synergy between Cdon mutation and in utero EtOH exposure occurred during Nodal-dependent specification of the PCP and that reduction of Nodal target gene expression required a combination of mutation and teratogen

Summary

Introduction

Many common structural birth defects appear to arise from a complex and ill-defined combination of genetic and environmental factors (Krauss and Hong, 2016). Consistent with the notion that a threshold of HH signaling activity is rate-limiting in midline patterning, genetic removal of one copy of the negative pathway regulator, Ptch, rescued 129S6 Cdon-/- mice from EtOH-induced HPE (Hong and Krauss, 2013). EtOH inhibits Activin/Nodal pathway signaling in vitro in mouse epiblast stem cells (mEpiSCs), which have transcriptional and functional properties that resemble APS cells (Kojima et al, 2014; Tsakiridis et al, 2014) Together these results argue that, unexpectedly, Cdon mutation and fetal alcohol synergize to induce HPE by interfering with Nodal signaling. These results illuminate mechanisms of gene-environment interaction in a high fidelity, multifactorial model of a common birth defect

Results

Discussion

Materials and methods

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