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Bone phenotype of P2X4 receptor knockout mice: implication of a P2X7 receptor mutation?

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Abstract

Transgenic and knockout animal models are widely used to investigate the role of receptors, signaling pathways, and other peptides and proteins. Varying results are often published on the same model from different groups, and much effort has been put into understanding the underlying causes of these sometimes conflicting results. Recently, it has been shown that a P2X4R knockout model carries a so-called passenger mutation in the P2X7R gene, potentially affecting the interpretation of results from studies using this animal model. We therefore report this case to raise awareness about the potential pitfalls using genetically modified animal models, especially within P2 receptor research. Although purinergic signaling has been recognized as an important contributor to the regulation of bone remodeling, the process that maintains the bone quality during life, little is known about the role of the P2X4 receptor (P2X4R) in regulation of bone remodeling in health and disease. To address this, we analyzed the bone phenotype of P2rx4tm1Rass (C57BL/6J) knockout mice and corresponding wildtype using microCT and biomechanical testing. Overall, we found that the P2X4R knockout mice displayed improved bone microstructure and stronger bones in an age- and gender-dependent manner. While cortical BMD, trabecular BMD, and bone volume were higher in the 6-month-old females and 3-month-old males, this was not the case for the 3-month-old females and the 6-month-old males. Bone strength was only affected in the females. Moreover, we found that P2X4R KO mice carried the P2X7 receptor 451P wildtype allele, whereas the wildtype mice carried the 451L mutant allele. In conclusion, this study suggests that P2X4R could play a role in bone remodeling, but more importantly, it underlines the potential pitfalls when using knockout models and highlights the importance of interpreting results with great caution. Further studies are needed to verify any specific effects of P2X4R on bone metabolism.

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Funding

The research was performed for internal funds from the Capitol Region of Denmark.

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Authors and Affiliations

  1. Department of Clinical Biochemistry, Rigshospitalet, Copenhagen, Denmark

    Maria Ellegaard, Tanja Hegner & Niklas Rye Jørgensen

  2. Institute of Clinical Research, University of Southern Denmark, Odense, Denmark

    Ming Ding

  3. Department of Orthopaedic Surgery and Traumatology, Odense University Hospital, Odense, Denmark

    Ming Ding

  4. IGF, Univ Montpellier, CNRS, INSERM, Montpellier, France

    Lauriane Ulmann

  5. LabEx ICST, Montpellier, France

    Lauriane Ulmann

  6. Institute of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

    Niklas Rye Jørgensen

Authors
  1. Maria Ellegaard
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  2. Tanja Hegner
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  3. Ming Ding
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  4. Lauriane Ulmann
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  5. Niklas Rye Jørgensen
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Contributions

Creating and providing animal model: LU. Planning of study: MEL, TH, and NRJ. Performing experiments: MEL and TH. Data analysis and interpretation: MEL, MD, and NRJ. Drafting manuscript: MEL. Critically reviewing, revising, and approving manuscript: MEL, TH, MD, LU, and NRJ.

Corresponding author

Correspondence to Niklas Rye Jørgensen.

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Conflicts of interest

Ellegaard M declares that she has no conflict of interest. Hegner T declares that she has no conflict of interest. Ding M declares that he has no conflict of interest. Ulmann L declares that she has no conflict of interest. Jørgensen NR declares that he has no conflict of interest.

Ethical approval

This article does not contain any studies with human participants performed by any of the authors. Animal studies were approved by the Danish Animal Experiments Inspectorate (License number 2017-15-0202-00086).

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Ellegaard, M., Hegner, T., Ding, M. et al. Bone phenotype of P2X4 receptor knockout mice: implication of a P2X7 receptor mutation?. Purinergic Signalling 17, 241–246 (2021). https://doi.org/10.1007/s11302-021-09784-9

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  • DOI: https://doi.org/10.1007/s11302-021-09784-9

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