# Ameloblastin Amphipathic Helix Motif mediates Ameloblast Polarization and Prismatic Enamel Formation via a RhoA Signaling Pathway

## TL;DR

This study shows how a specific protein motif in ameloblastin helps form tooth enamel structure through cell polarization and RhoA signaling.

## Contribution

The study identifies a novel signaling role of the amphipathic helix motif in ameloblastin for enamel prismatic structure formation.

## Key findings

- Deleting hydrophobic residues in the Ambn AH motif leads to hypo-mineralized enamel without prismatic structure.
- Mutant ameloblasts show loss of cell polarity and mis-localization of polarity markers like Pard3 and Claudin-1.
- AmbnΔL76-P86 mutants exhibit nuclear localization of β-catenin and p-Smad2/3 with reduced RhoA signaling.

## Abstract

Ameloblastin (Ambn) is a tooth-specific multifunctional protein crucial for enamel biomineralization and its prismatic structure. To examine the function of the evolutionarily conserved cell-binding Ambn amphipathic helix (AH) motif, we deleted the hydrophobic residues within Ambn AH motif in genetically engineered mice. Enamel in the homozygous (AmbnΔL76-P86) mutants had normal thickness but was hypo-mineralized and lacked prismatic structure. Micro-CT analysis using conventional neural network revealed loss in mineral density and a delay in the initiation of secretory stage of amelogenesis. Ameloblasts in the mutants were stunted and exhibited loss of cell polarity, as demonstrated by the mis-localization of Pard3, Claudin-1 and GM130 immunosignals. In the AmbnΔL76-P86 mutants, a loss of Ambn-ameloblast distal membrane interaction was observed, with nuclear localization of β-catenin and p-Smad2/3, and a decrease in RhoA immunolabeling intensity. Our study demonstrates a novel signaling role of Ambn AH motif in mediating cell polarization and forming enamel prismatic structure.