ID | 32960 |
FullText URL | |
Author |
Ris, Hans
Murshid, Sakhr A.
Yamamoto, Teruko Takano
Lim, Soo-Siang
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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
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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
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Publication Title |
Microscopy and Microanalysis
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Volume | volume13
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Issue | issue2
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Start Page | 108
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End Page | 117
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Content Type |
Journal Article
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language |
English
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Refereed |
True
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DOI | |
Web of Science KeyUT | |
Submission Path | materials_science/2
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