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The spontaneous mouse mutant low set ears (Lse) is caused by tandem duplication of Fgf3 and Fgf4

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Abstract

The external ear develops from an organized convergence of ventrally migrating neural crest cells into the first and second branchial arches. Defects in external ear position are often symptomatic of complex syndromes such as Apert, Treacher-Collins, and Crouzon Syndrome. The low set ears (Lse) spontaneous mouse mutant is characterized by the dominant inheritance of a ventrally shifted external ear position and an abnormal external auditory meatus (EAM). We identified the causative mutation as a 148 Kb tandem duplication on Chromosome 7, which includes the entire coding sequences of Fgf3 and Fgf4. Duplications of FGF3 and FGF4 occur in 11q duplication syndrome in humans and are associated with craniofacial anomalies, among other features. Intercrosses of Lse-affected mice revealed perinatal lethality in homozygotes, and Lse/Lse embryos display additional phenotypes including polydactyly, abnormal eye morphology, and cleft secondary palate. The duplication results in increased Fgf3 and Fgf4 expression in the branchial arches and additional discrete domains in the developing embryo. This ectopic overexpression resulted in functional FGF signaling, demonstrated by increased Spry2 and Etv5 expression in overlapping domains of the developing arches. Finally, a genetic interaction between Fgf3/4 overexpression and Twist1, a regulator of skull suture development, resulted in perinatal lethality, cleft palate, and polydactyly in compound heterozygotes. These data indicate a role for Fgf3 and Fgf4 in external ear and palate development and provide a novel mouse model for further interrogation of the biological consequences of human FGF3/4 duplication.

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Data availability

All data, including microCT image datasets, are available upon request.

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Acknowledgements

The authors thank the following for their assistance, expertise, information on strain history and/or helpful feedback on the manuscript: Bo Chang, Norm Hawes, Michelle Curtain, and Kevin Peterson. This work was supported by NIH Grants OD021325 (L.G.R. and D.E.B), EY015073 (L.R.D.), DE020052 (S.A.M. and L.R.D.)

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Author notes

  1. Alana Luzzio, Sarah Edie and Kristina Palmer have contributed equally to this work.

Authors and Affiliations

  1. The Jackson Laboratory, Bar Harbor, ME, USA

    Alana Luzzio, Sarah Edie, Kristina Palmer, L. Brianna Caddle, Rachel Urban, Leslie O. Goodwin, Ian C. Welsh, Laura G. Reinholdt, David E. Bergstrom, Leah Rae Donahue & Stephen A. Murray

  2. Departments of Oral & Craniofacial Sciences and Pediatrics, University of Missouri-Kansas City, Kansas City, MO, USA

    Timothy C. Cox

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  1. Alana Luzzio
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Contributions

SAM and LRD conceived and designed the experiments; AL, SE, KP, and LBC executed experiments, assembled figures, and analyzed results; IW helped design experiments, analyze data, and interpret results; RU and LOG generated and analyzed ddPCR results; LGR and DEB provided gene discovery support and analysis; TCC conducted microCT experiments and analyzed results; AL and SAM wrote the manuscript. All authors reviewed the manuscript and provided edits and feedback.

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Correspondence to Stephen A. Murray.

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The corresponding author, Stephen A. Murray, is an editor-in-chief of the journal. There are no other competing interests.

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Luzzio, A., Edie, S., Palmer, K. et al. The spontaneous mouse mutant low set ears (Lse) is caused by tandem duplication of Fgf3 and Fgf4. Mamm Genome 34, 453–463 (2023). https://doi.org/10.1007/s00335-023-09999-8

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