title: Increasing participation of sclerostin in postnatal bone development, revealed by three-dimensional immunofluorescence morphometry.
abstract: Confocal immunofluorescence tiling imaging revealed the spatio-temporal distributions of osterix and sclerostin in femurs from 3-day-old, 2-week-old and 4-week-old rats to be reciprocally exclusive at the tissue level. Further quantitative three-dimensional immuno fluorescence morphometry demonstrated the increasing distribution of sclerostin in the osteocytic lacuno-canalicular system specifically in diaphysis, which paralleled the cooperative participation and depletion of osterix and β-catenin in adjacent periosteum cells. Treating MC3T3-E1 cells with BIO (a GSK3 inhibitor) induced the stabilization of β-catenin and nuclear translocation of osterix, and negatively regulated osteocalcin/BGLAP and Dmp1. These results collectively demonstrate that the increasing distribution of sclerostin in diaphyseal cortical bone appears to be involved in the attenuation of osterix and β-catenin in adjacent periosteum cells, thus possibly contributing to osteoblast maturation and reducing the osteoblast formation at this bone site. Our confocal microscopy-based imaging analyses provide a comprehensive and detailed view of the spatio-temporal distribution of sclerostin, β-catenin and osterix at the tissue to subcellular level in a coherent manner, and uncovered their spatio-temporal cooperation in postnatal bone development, thus providing evidence that they link skeletogenic growth and functional bone development.
authors: Section of Oral Pathology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Japan.
authors: Takashi
authors: T
authors: Watanabe
authors: Yoshihiro
authors: Y
authors: Tamamura
authors: Akiyoshi
authors: A
authors: Hoshino
authors: Yuji
authors: Y
authors: Makino
authors: Hiroshi
authors: H
authors: Kamioka
authors: Teruo
authors: T
authors: Amagasa
authors: Akira
authors: A
authors: Yamaguchi
authors: Tadahiro
authors: T
authors: Iimura
mesh_terms: D000375:Aging / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001846:Bone Development / Q000187:drug effects; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D002452:Cell Count; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D018483:Diaphyses / Q000166:cytology / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D005269:Femur / Q000166:cytology / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D005455:Fluorescent Antibody Technique / Q000379:methods*; D005819:Genetic Markers; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007211:Indoles / Q000494:pharmacology; D008297:Male; D051379:Mice; D018613:Microscopy, Confocal; D010011:Osteocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010091:Oximes / Q000494:pharmacology; D055550:Protein Stability / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D000076182:Sp7 Transcription Factor; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000187:drug effects; D051176:beta Catenin / Q000378:metabolism
chemical_list: C483321:6-bromoindirubin-3'-oxime; D019485:Bone Morphogenetic Proteins; D050659:Core Binding Factor Alpha 1 Subunit; D005819:Genetic Markers; D007211:Indoles; D010091:Oximes; C493716:Runx2 protein, mouse; C486031:Sost protein, rat; D000076182:Sp7 Transcription Factor; C479466:Sp7 protein, mouse; C495181:Sp7 protein, rat; D014157:Transcription Factors; D051176:beta Catenin