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Journal of Electron Microscopy Advance Access published online on June 23, 2009
Journal of Electron Microscopy, doi:10.1093/jmicro/dfp032
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

FGF23 is mainly synthesized by osteocytes in the regularly distributed osteocytic lacunar canalicular system established after physiological bone remodeling

Sobhan Ubaidus1,2, Minqi Li1, Sara Sultana1,2, Paulo Henrique Luiz de Freitas1,2, Kimimitsu Oda1,3, Takeyasu Maeda1,4, Ritsuo Takagi2 and Norio Amizuka1,5*

1 Center for Transdisciplinary Research
2 Division of Oral and Maxillofacial Surgery
3 Division of Biochemistry
4 Division of Oral Anatomy, Graduate School of Medical and Dental Sciences, Niigata University, Niigata 951-8514, Japan
5 Division of Oral Health Science, Hokkaido University Graduate School of Dental Medicine, Sapporo 060-8586, Japan

* To whom correspondence should be addressed. E-mail: amizuka{at}den.hokudai.ac.jp

This study aimed to evaluate whether the immunolocalization of fibroblast growth factor (FGF) 23 and dentin matrix protein 1 (DMP1) is associated with the spatial regularity of the osteocyte lacunar canalicular system(s) (OLCS). Femora of 12-weeks-old male ICR mice were fixed with 4% paraformaldehyde, decalcified with a 10% EDTA solution and then embedded in paraffin. We have devised a triple staining procedure that combines silver impregnation, alkaline phosphatase (ALPase) immunohistochemistry and tartrate-resistant acid phosphatase (TRAPase) enzyme histochemistry on a single paraffin section. This procedure permitted the visualization of ALPase-positive plump osteoblasts and several TRAPase-positive osteoclasts on those bone matrices featuring irregularly arranged OLCS, and of ALPase-positive bone lining cells on the bone matrix displaying the well-arranged OLCS. As observations proceeded from the metaphysis toward the diaphysis, the endosteal cortical bone displayed narrower bands of calcein labeling, accompanied by increased regularity of the OLCS. This implies that the speed of bone deposition during bone remodeling would affect the regularity of the OLCS. While DMP1 was evenly localized in all regions of the cortical bones, FGF23 was more abundantly localized in osteocytes of cortical bones with regularly arranged OLCS. In cortical bones, the endosteal area featuring regular OLCS exhibited more intense FGF23 immunoreaction when compared to the periosteal region, which tended to display irregular OLCS. In summary, FGF23 appears to be synthesized principally by osteocytes in the regularly distributed OLCS that have been established after bone remodeling.

Keywords     FGF23, osteocyte, DMP1, bone remodeling, immunohistochemistry

Received      3 March 2009, accepted 22 May 2009


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