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10.08.2022 | Original Article

Deletion of Tfam in Prx1-Cre expressing limb mesenchyme results in spontaneous bone fractures

verfasst von: Hiroki Yoshioka, Shingo Komura, Norishige Kuramitsu, Atsushi Goto, Tomoka Hasegawa, Norio Amizuka, Takuya Ishimoto, Ryosuke Ozasa, Takayoshi Nakano, Yuuki Imai, Haruhiko Akiyama

Erschienen in: Journal of Bone and Mineral Metabolism | Ausgabe 5/2022

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Abstract

Introduction

Osteoblasts require substantial amounts of energy to synthesize the bone matrix and coordinate skeleton mineralization. This study analyzed the effects of mitochondrial dysfunction on bone formation, nano-organization of collagen and apatite, and the resultant mechanical function in mouse limbs.

Materials and methods

Limb mesenchyme-specific Tfam knockout (Tfam^f/f;Prx1-Cre: Tfam-cKO) mice were analyzed morphologically and histologically, and gene expressions in the limb bones were assessed by in situ hybridization, qPCR, and RNA sequencing (RNA-seq). Moreover, we analyzed the mitochondrial function of osteoblasts in Tfam-cKO mice using mitochondrial membrane potential assay and transmission electron microscopy (TEM). We investigated the pathogenesis of spontaneous bone fractures using immunohistochemical analysis, TEM, birefringence analyzer, microbeam X-ray diffractometer and nanoindentation.

Results

Forelimbs in Tfam-cKO mice were significantly shortened from birth, and spontaneous fractures occurred after birth, resulting in severe limb deformities. Histological and RNA-seq analyses showed that bone hypoplasia with a decrease in matrix mineralization was apparent, and the expression of type I collagen and osteocalcin was decreased in osteoblasts of Tfam-cKO mice, although Runx2 expression was unchanged. Decreased type I collagen deposition and mineralization in the matrix of limb bones in Tfam-cKO mice were associated with marked mitochondrial dysfunction. Tfam-cKO mice bone showed a significantly lower Young’s modulus and hardness due to poor apatite orientation which is resulted from decreased osteocalcin expression.

Conclusion

Mice with limb mesenchyme-specific Tfam deletions exhibited spontaneous limb bone fractures, resulting in severe limb deformities. Bone fragility was caused by poor apatite orientation owing to impaired osteoblast differentiation and maturation.

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Titel: Deletion of Tfam in Prx1-Cre expressing limb mesenchyme results in spontaneous bone fractures
verfasst von: Hiroki Yoshioka
Shingo Komura
Norishige Kuramitsu
Atsushi Goto
Tomoka Hasegawa
Norio Amizuka
Takuya Ishimoto
Ryosuke Ozasa
Takayoshi Nakano
Yuuki Imai
Haruhiko Akiyama
Publikationsdatum: 10.08.2022
Verlag: Springer Nature Singapore
Erschienen in: Journal of Bone and Mineral Metabolism / Ausgabe 5/2022
Print ISSN: 0914-8779
Elektronische ISSN: 1435-5604
DOI: https://doi.org/10.1007/s00774-022-01354-2

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Deletion of Tfam in Prx1-Cre expressing limb mesenchyme results in spontaneous bone fractures