ARHGAP29 Mutation Is Associated with Abnormal Oral Epithelial Adhesions

B J Paul; J C Sharp; K Palmer; C H Pratt; S A Murray; M Dunnwald

doi:10.1177/0022034517726079

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ARHGAP29 Mutation Is Associated with Abnormal Oral Epithelial Adhesions

Journal article

Open access

Peer reviewed

ARHGAP29 Mutation Is Associated with Abnormal Oral Epithelial Adhesions

B J Paul, J C Sharp, K Palmer, C H Pratt, S A Murray and M Dunnwald

Journal of dental research, Vol.96(11), pp.1298-1305

10/2017

DOI: 10.1177/0022034517726079

PMCID: PMC5613885

PMID: 28817352

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https://europepmc.org/articles/pmc5613885View

Published (Version of record) Open Access

Abstract

Nonsyndromic cleft lip and/or palate (NSCL/P) is a prevalent birth defect of complex etiology. Previous studies identified mutations in ARHGAP29 associated with an increased risk for NSCL/P. To investigate the effects of ARHGAP29 in vivo, we generated a novel murine allele by inserting a point mutation identified in a patient with NSCL/P. This single-nucleotide variation of ARHGAP29 translates to an early nonsense mutation (K326X), presumably resulting in loss-of-function (LoF). Embryos from Arhgap29 intercrosses were harvested at various time points. No homozygous Arhgap29 animals were found in the 45 analyzed litters, assessed as early as embryonic day 8.5 (e8.5). Coronal sectioning of e13.5 and e14.5 heads revealed that 59% of Arhgap29 mice ( n = 37) exhibited improper epithelial contact between developing oral structures, while none were observed in wild types ( n = 10). In addition, Arhgap29 embryos exhibited a significantly higher percentage of maxillary epithelium in contact with mandibular epithelium. Immunofluorescent analyses of the periderm and oral adhesions revealed the presence of Arhgap29 in periderm cells. These cells were p63 negative, keratin 17 positive, and keratin 6 positive and present at sites of adhesion, although occasionally disorganized. Oral adhesions did not appear to impair palatogenesis, as all analyzed Arhgap29 embryos showed confluent palatal mesenchyme and epithelium at e18.5 ( n = 16), and no mice were found with a cleft at birth. Collectively, our data demonstrate that ARHGAP29 is required for embryonic survival and that heterozygosity for LoF variants of Arhgap29 increases the incidence and length of oral adhesions at a critical time point during orofacial development. In conclusion, we validate the LoF nature of the human K326X mutation in vivo and reveal a previously unknown effect of Arhgap29 in murine craniofacial development.

Phenotype

Genome-Wide Association Study

Exons

Cleft Lip - embryology

Humans

Cleft Palate - embryology

Cleft Palate - genetics

Codon, Nonsense

Embryonic Development

Mice, Inbred Strains

Cell Adhesion

Blotting, Western

Point Mutation

Cleft Lip - genetics

Animals

Codon

Alleles

Mice

Polymorphism, Single Nucleotide

GTPase-Activating Proteins - genetics

Details

Title: Subtitle: ARHGAP29 Mutation Is Associated with Abnormal Oral Epithelial Adhesions
Creators: B J Paul - 1 Department of Anatomy and Cell Biology, The University of Iowa, Iowa City, IA, USA
J C Sharp - 2 The Jackson Laboratory, Bar Harbor, ME, USA
K Palmer - 2 The Jackson Laboratory, Bar Harbor, ME, USA
C H Pratt - 2 The Jackson Laboratory, Bar Harbor, ME, USA
S A Murray - 2 The Jackson Laboratory, Bar Harbor, ME, USA
M Dunnwald - 1 Department of Anatomy and Cell Biology, The University of Iowa, Iowa City, IA, USA
Resource Type: Journal article
Publication Details: Journal of dental research, Vol.96(11), pp.1298-1305
DOI: 10.1177/0022034517726079
PMID: 28817352
PMCID: PMC5613885
NLM abbreviation: J Dent Res
ISSN: 0022-0345
eISSN: 1544-0591
Publisher: United States
Grant note: R37 DE008559 / NIDCR NIH HHS U01 DE020052 / NIDCR NIH HHS R01 AR067739 / NIAMS NIH HHS
Language: English
Date published: 10/2017
Academic Unit: Anatomy and Cell Biology; Craniofacial Anomalies Research Center
Record Identifier: 9984025362102771

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