Loss of ninein interferes with osteoclast formation and causes premature ossification
- Molecular, Cellular and Developmental Biology, Centre de Biologie Intégrative, UMR5077, CNRS & Université Paul Sabatier, France
- Institut de Pharmacologie et de Biologie Structurale, UMR5089, CNRS & Université Paul Sabatier, France
- CIRIMAT, UMR5085, CNRS & Université Paul Sabatier, France
- International Research Project CNRS “MAC-TB/HIV”, France
Figures
Figure 1
Ninein knock-out mice display reduced litter size upon a zygotic genotype.
(A) Litter size comparisons following crossings among control or ninein-deleted animals at birth. Blue-colored bars: live pups, orange-colored bars: dead pups. Crossing of males and females with …
Figure 2
Advanced ossification in ninein-deleted mice.
(A) Whole skeleton preparations of E18.5 embryos using Alcian blue staining for cartilage and Alizarin red staining for mineralized bone. Left: control (heterozygous), right: ninein del/del embryo. …
Figure 3 with 1 supplement
Advanced endochondral ossification in ninein-deleted mice.
(A) Whole skeleton preparation of E16.0 embryos, stained for mineralized bone using Alizarin red. Left, control heterozygous, right: ninein del/del embryo. Despite an overall similar staining, …
Figure 3—figure supplement 1
Early endochondral ossification is present at multiple sites of bone formation.
Skeleton staining of 21.5dpc pups using Alcian blue and Alizarin red. (A) Forelimbs and (B) hindlimbs from control (heterozygous) and ninein-deleted neonates. Note the advanced ossification in …
Figure 4
Early intramembranous ossification in ninein-deleted mice leads to premature suture closure.
(A, B) Coronal sections through E16.5 heads of control and ninein-deleted embryos (e, eye; f, frontal; is, interfrontal suture; md, mandible; pa, palatal; t, tongue). (A) Von Kossa staining revealed …
Figure 5
Reduced osteoclast density affects long bone internal structure at E18.5.
(A) Histology of E16.5 heterozygous Nintm1a mice revealed high expression of β-galactosidase (blue) in multinucleated cells (arrows). (B) These β-galactosidase-positive syncytia line close to …
Figure 6 with 1 supplement
Reduced cell fusion of osteoclast precursors during osteoclastogenesis.
(A) Focus on mandible development from sections of E14.5 control and ninein-deleted embryos. Examination of TRAP-positive cells within the mandible area, revealing the presence of mononucleated …
Figure 6—figure supplement 1
Early osteoclastogenesis from erythro-myeloid progenitors.
(A, B) Early bone development at E14.5, examined by von-Kossa-staining of longitudinal sections of whole control and ninein-deleted embryos. The absence of silver deposit in either group indicates …
Figure 7 with 1 supplement
Reduced cell fusion of osteoclast precursors from cultured bone marrow cells.
(A) TRAP staining of osteoclasts from adult mouse bone marrow at day 3 of culture, revealing size differences between controls and ninein del/del. (B) Fusion of precursors cells from bone marrow …
Figure 7—figure supplement 1
Time course of osteoclast fusion, and rates of proliferation, viability, and cell adhesion of precursor cells.
(A) The surface of TRAP-positive syncytia (≥2 nuclei) was determined on entire coverslips (Ø 12 mm), and categories of syncytia up to 5000 μm2 and above were established. Cell numbers for 3 …
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Figure 7—figure supplement 1—source data 1
Immunoblots of osteoblast precursors from control and ninein del/del mice, one day before fusion, probed with antibodies against poly-(ADP-ribose)-polymerase (PARP) and cleaved caspase 3, and probed with antibody against Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a loading control.
- https://cdn.elifesciences.org/articles/93457/elife-93457-fig7-figsupp1-data1-v2.pdf
Figure 8
Abnormal centriole clustering in osteoclast cultures from ninein-deleted animals.
(A) Double immunofluorescence of neonatal osteoclast progenitors for ninein (nin) and centrin (cen), (B) α and γ-tubulin, and (C) γ-tubulin and dynactin-subunit p150. Controls (upper images) and …
Additional files
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Supplementary file 1
Forelimb and hindlimb bone characteristics in control and ninein-deleted mice.
Table, indicating the lengths of various bones from embryos at E16.5 and E18.5, from control and ninein del/del embryos.
- https://cdn.elifesciences.org/articles/93457/elife-93457-supp1-v2.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/93457/elife-93457-mdarchecklist1-v2.pdf
