In:
Microscopy and Microanalysis, Oxford University Press (OUP), Vol. 13, No. 2 ( 2007-04), p. 108-117
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 (18α-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.
Type of Medium:
Online Resource
ISSN:
1431-9276
,
1435-8115
DOI:
10.1017/S143192760707016X
Language:
English
Publisher:
Oxford University Press (OUP)
Publication Date:
2007
detail.hit.zdb_id:
1481716-0
SSG:
11
SSG:
12
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