Glycogen synthase kinase 3 alpha/beta deletion induces precocious growth plate remodeling in mice

Abstract

Glycogen synthase kinase (GSK) 3 acts to negatively regulate multiple signaling pathways, including canonical Wnt signaling. The two mammalian GSK3 proteins (alpha and beta) are at least partially redundant. While Gsk3a KO mice are viable and display a metabolic phenotype, abnormal neuronal development, and accelerated aging, Gsk3b KO animals die late in embryogenesis or at birth. Selective Gsk3b KO in bone delays development of some bones, whereas cartilage-specific Gsk3b KO mice are normal except for elevated levels of GSK3A protein. However, the collective role of these two GSK3 proteins in cartilage was not evaluated. To address this, we generated tamoxifen-inducible, cartilage-specific Gsk3a/Gsk3b KO (described as “cDKO”) in juvenile mice and investigated their skeletal phenotypes. We found that cartilage-specific Gsk3a/Gsk3b deletion in young, skeletally immature mice causes precocious growth plate (GP) remodeling, culminating in shorter long bones and hence, growth retardation. These mice exhibit inefficient breathing patterns at later stages and fail to survive. The disrupted GP in cDKO mice showed progressive loss of cellular and proteoglycan components, and immunostaining for SOX9, while BGLAP (osteocalcin) and COL2A1 increased. In addition, we observed increased osteoclast recruitment and cell apoptosis. Surprisingly, changes in articular cartilage of cDKO mice were mild compared with the GP, signifying differential regulation of articular cartilage vs GP tissues. Taken together, these findings emphasize a crucial role of two GSK3 proteins in skeletal development, in particular in the maintenance and function of GP.

Key Messages

• Both GSK3 genes, together, are crucial regulators of growth plate remodeling.

• Cartilage-specific deletion of both GSK3 genes causes skeletal growth retardation.

• Deletion of both GSK3 genes decreases Sox9 levels and promotes chondrocyte apoptosis.

• Cartilage-specific GSK3 deletion in juvenile mice culminates in premature lethality.

• GSK3 deletion exhibits mild effects on articular cartilage compared to growth plate.

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Data Availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We thank the following funding sources for financial support. S.K.B. received a postdoctoral fellowship award from CONNECT! NSERC CREATE Program in Soft Connective Tissue Regeneration/Therapy. S.K.B. and C.P. were also supported by the Transdisciplinary Training Award from Collaborative Program in Musculoskeletal Health Research (CMHR) at Western’s Bone and Joint Institute. F.B holds the Canada Research Chair in Musculoskeletal Research and is the recipient of a Foundation Grant from the Canadian Institutes of Health Research (Grant #332438). We thank all members of our lab for valuable discussions and encouragement, in particular Julia Bowering for tissue sectioning services.

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Conceptualization and methodology, S.K.B. and F.B.; investigation, S.K.B., D.B., C.P.; analyses, S.K.B., C.P. and F.B.; manuscript preparation and review, S.K.B., D.B., J.R.W., and F.B. All authors approved submission of manuscript.

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Correspondence to Frank Beier.

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All experimental mice were handled in accordance with the guidelines from the Canadian Council on Animal Care, and experiments were approved by the Animal Use Subcommittee at The University of Western Ontario.

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Bali, S.K., Bryce, D., Prein, C. et al. Glycogen synthase kinase 3 alpha/beta deletion induces precocious growth plate remodeling in mice. J Mol Med (2021). https://doi.org/10.1007/s00109-021-02049-3

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Keywords

  • Growth plate
  • GSK3
  • Articular cartilage
  • Bone length
  • Skeletal development
  • Longitudinal growth