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. 2022 Oct 1;12:16477. doi: 10.1038/s41598-022-20684-9

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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FIB-bSEM of 7-week Acp4^+/+ and Acp4^R110C/R110C Mandibular Incisors. To compare the ultrastructure of early enamel formed in Acp4^+/+ and Acp4^R110C/R110C mice, sites are sampled (1) after dentin mineralizes and before enamel secretion begins (A, F; Level 0.92), (2) just after enamel secretion starts (B, G; level 1.11), (3) transition from formation of the initial enamel layer to development of Tomes processes (C, H; level 1.20), and (4) early inner enamel layer formation (D, I; Level 1.33). (E) A brighter version of image of (D). Low magnification (mag) images (A–E) (2500×) with mag bar in (B); high mag images (F–I) (35,000×) with mag bar in (G). In Acp4^+/+ mice, initial enamel mineral ribbons appear to elongate away from the tips and sides of mineralized collagen fibers at the DEJ (A, B). This process continues as the enamel prongs map out the Tomes processes (C) and continue on with formation of rod and interrod areas typical of the inner enamel layer (D). In Acp4^R110C/R110C mice, initial enamel ribbons start to form sporadically (F, G), but ameloblasts are unable to elongate them to create an initial layer, enamel prongs or Tomes processes (G, H), thereby negating any further normal development of an enamel layer. Noticeably, those initial ribbons are not associated with the ameloblast cell membrane. The mineral crystals formed in the absence of normal ACP4 appear thin, short in length and closely compressed close to each other (I). Am, ameloblasts; d, dentin; e, enamel; Od, odontoblasts; pd: predentin.