The ectodermal loss of ARHGAP29 alters epithelial morphology and organization and disrupts murine palatal development

Adelizzi, E.; Rhea, L.; Mitvalsky, C.; Pek, S.; Doolittle, B.; Dunnwald, M.

doi:10.1101/2025.03.11.642653

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bioRxiv · 10.1101/2025.03.11.642653

The ectodermal loss of ARHGAP29 alters epithelial morphology and organization and disrupts murine palatal development

Adelizzi, E.·Rhea, L.·Mitvalsky, C.·Pek, S.·Doolittle, B.·Dunnwald, M.

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Abstract

Orofacial clefts, including cleft palate (CP), are among the most common types of birth defects. CP specifically, results from a failure of palatal shelf fusion during development. Previous studies have shown that mutations in RhoA GTPase Activating Protein 29 (ARHGAP29) are linked to CP, yet the role and tissue-specific requirements for ARHGAP29 during palatogenesis remain unknown. Here, we use tissue-specific deletion of Arhgap29 in mice to provide the first direct evidence that ARHGAP29 is essential for proper palatal elevation and fusion. We demonstrate that ectodermal conditional loss of Arhgap29 induces a significant delay in the fusion of palatal shelves at embryonic (E) day 14.5 and an incomplete yet significantly penetrant cleft palate at E18.5 - neither of which are observed when Arhgap29 is lost later in development using K14-Cre. Phenotypic analyses of palatal shelves at E14.5 reveal a disorganized and thicker epithelium at the tip of the shelves. Loss of Arhgap29 increases palate epithelial cell area and upregulates alpha-smooth muscle actin and phospho-myosin regulatory light chain implicating cell morphology and contractility as drivers of CP. Summary statementThis study in mice is the first direct evidence that ARHGAP29 is essential for proper palatal elevation and fusion. Loss of Arhgap29 alters oral epithelial morphology and upregulates contractility proteins.

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Adelizzi, E., Rhea, L., Mitvalsky, C., Pek, S., Doolittle, B., Dunnwald, M.. 2025-03-13. The ectodermal loss of ARHGAP29 alters epithelial morphology and organization and disrupts murine palatal development. https://doi.org/10.1101/2025.03.11.642653

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