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Characterization of the congenital diaphragmatic hernia model in C57BL/6J fetal mice: a step toward lineage tracing experiments

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Abstract

Purpose

Lineage tracing is key to study the fate of individual cells and their progeny especially in developmental biology. To conduct these studies, we aimed to establish a reproducible model of CDH in the most commonly used genetic background strain that is C57BL/6J mice.

Methods

CDH was induced in C57BL/6J dams by maternal administration of nitrofen + bisdiamine at E8.5. Fetuses from olive oil-gavaged mothers served as controls. Lungs from CDH and control fetuses were compared for (1) growth via radial airspace count (RAC), mean linear intercept (MLI) and gene expression for Fgf10, Nrp1, and Ctnnb1; (2) maturation (Pdpn, Spc, Ager, Abca3, Eln, Acta2, Pdgfra) via gene and protein expression; (3) vascularization via gene and protein expression (CD31, Vegfa, Vegfr1/2, Epas1, Enos). Statistics: unpaired t-test or Mann–Whitney test.

Results

Nitrofen + bisdiamine administration resulted in 36% left-sided CDH (31% mortality). CDH fetuses had hypoplastic lungs and impaired growth (lower RAC, higher MLI, lower Fgf10, Nrp1, Ctnnb1), maturation (decreased Pdpn, Ager, Eln gene expression), and vascularization (decreased Cd31, Vegfr1/2; Epas1 and Enos). Lower protein expression was confirmed for PDPN, ELN and CD31.

Conclusion

Modeling CDH in C57BL/6J mouse fetuses is effective in reproducing the classical CDH hallmarks. This model will be critical for lineage tracing experiments.

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Data availability

All data supporting the findings of this study are available within the paper.

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Acknowledgements

This project was supported by the Canadian Institutes of Health Research (CIHR) Project Grant (175300), SickKids Congenital Diaphragmatic Hernia Fund (R00DH00000) at the Hospital for Sick Children, Toronto, Canada and the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)—466815475.

Author information

Authors and Affiliations

  1. Developmental and Stem Cell Biology Program, Peter Gilgan Centre for Research and Learning, The Hospital for Sick Children, Toronto, M5G 0A4, Canada

    Fabian Doktor, Rebeca Lopes Figueira, Kasra Khalaj, Aizah Ijaz, Matisse Blundell, Lina Antounians & Augusto Zani

  2. Division of General and Thoracic Surgery, The Hospital for Sick Children, Toronto, M5G 1X8, Canada

    Fabian Doktor, Rebeca Lopes Figueira, Kasra Khalaj, Aizah Ijaz, Matisse Blundell, Lina Antounians & Augusto Zani

  3. Department of Pediatric Surgery, University of Leipzig, 04109, Leipzig, Germany

    Fabian Doktor & Martin Lacher

  4. Department of Surgery, University of Toronto, Toronto, M5T 1P5, Canada

    Augusto Zani

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  1. Fabian Doktor
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Contributions

Conceptualization: FD, LA, AZ. Methodology: FD, RLF, KK, AI, MB. Visualization: FD, RLF, KK, AI, MB. Project administration: LA, AZ. Funding acquisition: FD, ML, AZ. Supervision: LA, ML, AZ. Writing—original draft preparation: FD. Writing—review and editing: LA, AZ.

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Correspondence to Augusto Zani.

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Doktor, F., Figueira, R.L., Khalaj, K. et al. Characterization of the congenital diaphragmatic hernia model in C57BL/6J fetal mice: a step toward lineage tracing experiments. Pediatr Surg Int 39, 296 (2023). https://doi.org/10.1007/s00383-023-05583-y

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