Osteocytes regulate senescence of bone and bone marrow
- Department of Orthopaedics, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, China
- Centre for Orthopaedic Translational Research, Medical School, University of Western Australia, Australia
- Perron Institute for Neurological and Translational Science, Australia
- Ningbo Institute of Life and Health Industry, University of Chinese Academy of Sciences, China
- Institute of Microsurgery on Extremities, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, China
Figures
Figure 1 with 1 supplement
DTAhet mice display partial osteocyte ablation.
(A, B) Hematoxylin–eosin staining of WT and DTAhet mice femur at 4 weeks (A) and quantification of the ratio of empty lacunae (arrows) (B) (n = 8–12 per group), indicating reduced osteocyte number …
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Figure 1—source data 1
DTAhet mice display partial osteocyte ablation.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
Osteocyte ablation has no impact on embryonic skeletal development.
(A) Whole-mount skeleton staining of WT and DTAhet mice at E19.0 by Alizarin red/Alcian blue. (B, C) Hematoxylin–eosin staining of WT and DTAhet mice femur at 13 weeks (B) and quantification of the …
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Figure 1—figure supplement 1—source data 1
DTAhet mice display partial osteocyte ablation.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig1-figsupp1-data1-v2.xlsx
Figure 2 with 2 supplements
Osteocyte ablation induces severe osteoporosis and kyphosis.
(A–C) Representative micro-computed tomography (µCT) reconstructive images of male WT and DTAhet mice femur at 4 weeks (A) and trabecular microstructural parameters (BMD, bone mineral density; …
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Figure 2—source data 1
Osteocyte ablation induces severe osteoporosis in male mice.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
Osteocyte ablation induces severe osteoporosis and kyphosis.
(A–C) Representative micro-computed tomography (µCT) reconstructive images of female WT and DTAhet mice femur at 4 weeks (A) and trabecular microstructural parameters (BMD, bone mineral density; …
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Figure 2—figure supplement 1—source data 1
Osteocyte ablation induces severe osteoporosis in female mice.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig2-figsupp1-data1-v2.xlsx
Figure 2—figure supplement 2
Osteocyte ablation induces severe osteoporosis and kyphosis.
(A) Gait analysis of normal step sequence ratio of male WT and DTAhet mice at 4 weeks (n = 6 per group). (B–D) Representative gait images and foot pattern of male WT and DTAhet mice (B) at 4 weeks …
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Figure 2—figure supplement 2—source data 1
Osteocyte ablation induces abnormal steps of mice.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig2-figsupp2-data1-v2.xlsx
Figure 3 with 1 supplement
Osteocyte ablation leads to severe sarcopenia and shorter lifespan.
(A, B) Gross images (A) and weight (B) of male WT and DTAhet mice at 4 weeks (n = 5–8 per group). (C) The ratio of gastrocnemius muscle weight in WT and DTAhet mice at 4 weeks (n = 3 per group). (D, …
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Figure 3—source data 1
Osteocyte ablation leads to severe sarcopenia and shorter lifespan.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Osteocyte ablation leads to severe sarcopenia and shorter lifespan.
(A, B) The quantification of muscle fiber number of WT and DTAhet mice at 4 weeks (A) and 13 weeks (B) (n = 5–6 per group). (C) RT-qPCR verification of Dmp1 expression of cortical bone and muscle (n …
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Figure 3—figure supplement 1—source data 1
Osteocyte ablation leads to severe sarcopenia.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig3-figsupp1-data1-v2.xlsx
Figure 4 with 1 supplement
Ablation of osteocytes alters mesenchymal lineage commitment and promotes osteoclastogensis.
(A, B) Goldner trichrome staining of male WT and DTAhet mice femur at 4 weeks (A) and histomorphometry analysis of osteoblast numbers (N.Ob/BS) (arrows) and osteoid-covered surface (OS/BS) (B) (n = …
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Figure 4—source data 1
Ablation of osteocytes alters mesenchymal lineage commitment and promotes osteoclastogensis.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig4-data1-v2.xlsx
Figure 4—figure supplement 1
Ablation of osteocytes alters mesenchymal lineage commitment and promotes osteoclastogensis.
(A) Selected osteogenesis-related Gene Ontology (GO) analysis of downregulated genes by osteocyte ablation. (B) Heatmap of significantly differentiated genes (fold change > 2.0-fold, WT FPKM > 10, …
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Figure 4—figure supplement 1—source data 1
Ablation of osteocytes alters mesenchymal lineage commitment and promotes osteoclastogensis.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig4-figsupp1-data1-v2.xlsx
Figure 5 with 3 supplements
Alteration of hematopoietic lineage commitment by osteocyte ablation.
(A, B) The Uniform Manifold Approximation and Projection (UMAP) plot of cells isolated from the bone marrow of 4-week WT and DTAhet mice and inferred cluster identity (A) and number of mRNA per cell …
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Figure 5—source data 1
Alteration of hematopoietic lineage commitment by osteocyte ablation.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig5-data1-v2.xlsx
Figure 5—figure supplement 1
Flow cytometry gating strategy.
(A) Flow cytometry of gating hematopoietic stem cell (HSC) (Lin-Sca1+c-Kit+), long-term HSC (LT-HSC) (Lin-Sca1+c-Kit+Flk2-), short-term HSC (ST-HSC) (Lin-Sca1+c-Kit+Flk2+), common myeloid …
Figure 5—figure supplement 2
Alteration of hematopoietic lineage commitment by osteocyte ablation.
(A, B) Representative images of flow cytometry (A) and analysis of proportions of hematopoietic stem cells (HSC) (B) of 4-week WT and DTAhet mice (n = 3–4 per group). (C, D) Representative image of …
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Figure 5—figure supplement 2—source data 1
Alteration of hematopoietic lineage commitment by osteocyte ablation.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig5-figsupp2-data1-v2.xlsx
Figure 5—figure supplement 3
Increased granulopoiesis after osteocyte ablation.
(A) The Uniform Manifold Approximation and Projection (UMAP) plot of neutrophils of 4-week WT and DTAhet mice and inferred subcluster identity. (B) Dot plot showing the scaled expression of selected …
Figure 6 with 1 supplement
Senescence of osteoprogenitors and myeloid lineage cells leads to the accelerated skeletal aging.
(A) Comparisons of total bone marrow cells senescence-associated secretory phenotype (SASP) score between 4-week WT and DTAhet mice. (B) Latent time of RNA velocity analysis of WT and DTAhet mice …
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Figure 6—source data 1
Senescence of osteoprogenitors and myeloid lineage cells leads to the accelerated skeletal aging.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig6-data1-v2.xlsx
Figure 6—figure supplement 1
Senescence of osteoprogenitors and myeloid lineage cells leads to the accelerated skeletal aging.
(A) SA-βGal staining of frozen sections of femur of WT and DTAhet mice. Scale bar, 20 µm. (B) Bar plot of Gene Ontology (GO) analysis of mesenchymal stem cell (MSC) cluster. (C, D) RT-qPCR analysis …
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Figure 6—figure supplement 1—source data 1
Osteocyte ablation induced MSC lineage towards adipogenesis.
- https://cdn.elifesciences.org/articles/81480/elife-81480-fig6-figsupp1-data1-v2.xlsx
Figure 7
Schematic diagram of osteocyte ablation-induced skeletal senescence.
Ablation of osteocytes induced senescence-associated secretory phenotype (SASP) accumulation in bone marrow osteoprogenitors and myeloid lineage cells, which altered mesenchymal stem cell (MSC) and …
Videos
Video 1
Representative movie showing movement in WT mice at 4 weeks.
Video 2
Representative movie showing movement defects in DTAhet mice at 4 weeks.
Video 3
Representative movie showing movement in WT mice at 13 weeks.
Video 4
Representative movie showing movement defects in DTAhet mice at 13 weeks.
Video 5
Representative movie showing movement in WT mice at 37 weeks.
Video 6
Representative movie showing movement defects in DTAhet mice at 37 weeks.
Additional files
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Supplementary file 1
Selected skeleton-related Gene Ontology (GO analysis).
- https://cdn.elifesciences.org/articles/81480/elife-81480-supp1-v2.xlsx
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Supplementary file 2
Gene sets involving neutrophil function and senescence-associated secretory phenotype (SASP).
- https://cdn.elifesciences.org/articles/81480/elife-81480-supp2-v2.xlsx
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MDAR checklist
- https://cdn.elifesciences.org/articles/81480/elife-81480-mdarchecklist1-v2.docx
