このエントリーをはてなブックマークに追加
ID 32960
FullText URL
Author
Ris, Hans
Murshid, Sakhr A.
Yamamoto, Teruko Takano
Lim, Soo-Siang
Abstract

The inaccessibility of osteocytes due to their embedment in the calcified bone matrix in vivo has precluded direct demonstration that osteocytes use gap junctions as a means of intercellular communication. In this article, we report successfully isolating primary cultures of osteocytes from chick calvaria, and, using anti-connexin 43 immunocytochemistry, demonstrate gap junction distribution to be comparable to that found in vivo. Next, we demonstrate the functionality of the gap junctions by (1) dye coupling studies that showed the spread of microinjected Lucifer Yellow from osteoblast to osteocyte and between adjacent osteocytes and (2) analysis of fluorescence replacement after photobleaching (FRAP), in which photobleaching of cells loaded with a membrane-permeable dye resulted in rapid recovery of fluorescence into the photobleached osteocyte, within 5 min postbleaching. This FRAP effect did not occur when cells were treated with a gap junction blocker (18 alpha-glycyrrhetinic acid), but replacement of fluorescence into the photobleached cell resumed when it was removed. These studies demonstrate that gap junctions are responsible for intercellular communication between adjacent osteocytes and between osteoblasts and osteocytes. This role is consistent with the ability of osteocytes to respond to and transmit signals over long distances while embedded in a calcified matrix.

Keywords
osteocytes
osteoblasts
gap junctions
intercellular communication
dye coupling
FRAP
Note
Published with permission from the copyright holder.
This is as published in Microscopy and Microanalysis , 2007 APR Vol.13 Issue.2 pp.108-117
Publisher URL: http://dx.doi.org/10.1017/S143192760707016X
Direct access to Thomson Web of Science record
Copyright © 2007 by Microscopy Society of America
Published Date
2008-09-21
Publication Title
Microscopy and Microanalysis
Volume
volume13
Issue
issue2
Start Page
108
End Page
117
Content Type
Journal Article
language
英語
Refereed
True
DOI
Web of Sience KeyUT
Submission Path
materials_science/2