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Citrate Transporter Expression and Localization: The Slc13a5Flag Mouse Model.

Jan C-C Hu1, Tian Liang2, Hong Zhang1

  • 1Department of Biologic and Materials Sciences & Prosthodontics, University of Michigan School of Dentistry, 1011 N University Ave., Ann Arbor, MI 48109, USA.

International Journal of Molecular Sciences
|July 29, 2025
PubMed
Summary
This summary is machine-generated.

We developed Slc13a5Flag reporter mice to track the sodium-citrate cotransporter (NaCT) during tooth development. This functional reporter shows NaCT is present on ameloblasts, suggesting citrate enters enamel via paracellular routes or bidirectional transport.

Keywords:
amelogenesisenamel mineralizationprotein localization

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Area of Science:

  • Biochemistry
  • Genetics
  • Developmental Biology

Background:

  • The sodium-citrate cotransporter (NaCT), encoded by the SLC13A5 gene, is vital for citrate transport in amelogenesis.
  • Mutations in SLC13A5 are linked to early infantile epileptic encephalopathy 25 and amelogenesis imperfecta.
  • Citrate is highly concentrated and mineral-bound in developing enamel.

Purpose of the Study:

  • To develop a reporter mouse model for precise localization of NaCT during tooth development.
  • To investigate the mechanism of citrate entry into developing enamel.

Main Methods:

  • Generation of Slc13a5Flag reporter mice expressing a C-terminally Flag-tagged NaCT.
  • Validation of the reporter mouse model using Sanger sequencing and phenotypic analysis.
  • Extensive characterization of tooth development via microscopy, in situ hybridization, and immunohistochemistry.

Main Results:

  • Slc13a5Flag reporter mice exhibited normal development and tooth formation.
  • NaCT-Flag was localized to the outer membranes of secretory and maturation-stage ameloblasts, with strong signals on the Tomes process.
  • The papillary layer and odontoblast outer membranes also showed NaCT-Flag staining, while ANK expression was absent in ameloblasts.

Conclusions:

  • The developed Slc13a5Flag reporter mice provide a reliable tool for studying NaCT localization and function.
  • NaCT's presence on ameloblast membranes suggests citrate enters enamel either paracellularly or via bidirectional NaCT transport.
  • These findings advance our understanding of citrate homeostasis in amelogenesis and related disorders.