Abstract
Polyclonal antisera to synthetic peptides matching sequences on the cytoplasmic regions of connexin-43, a gap junction protein first identified in rat heart, have been used to immunolabel gap junctions in the calvarial bone, maintained intact as in vivo, of 1- to 2-week-old rats. The specimens were examined in reflection and fluorescence modes by scanning laser confocal microscopy, and the numbers of gap junctions and their sizes estimated. The mean number of connexin-43 immunolabelled junctions per osteoblast (n = 65) was 15.3 (SD +/- 4.5). The mean length of 227 junctions, selected for the sharpness of the image of the fluorescent spot, was 0.67 micron (SD +/- 0.18; range 0.37-1.29 microns) and their mean area 0.26 micron2 (SD +/- 0.145; range 0.075-0.93 micron2); these probably fell within the upper half of the total size range. Gap junctions were detected between preosteoblasts, osteoblasts, osteocytes and chondrocytes, and between these juxtaposed cell types. In addition, connexin-43 immunolabelled junctions were found between some osteoclasts and overlying mononuclear cells at active sites of resorption.
MeSH Terms
Animals
Connexins
Intercellular Junctions/chemistry,ultrastructure
Membrane Proteins/analysis
Osteoblasts/chemistry,ultrastructure
Osteocytes/chemistry,ultrastructure
Rats
Rats, Sprague-Dawley
Skull/chemistry,ultrastructure
Chemicals
Connexins
Membrane Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Jones S J
Department of Anatomy and Developmental Biology, University College London, UK.
Gray C
Sakamaki H
Arora M
Boyde A
Gourdie R
Green C
References (26)
26 references, click to expand
-
Tight junctions between osteocyte processes.
Scand J Dent Res. 1973;81(4):339-41
PMID: 4518606
-
Cell-to-cell communication of osteoblasts.
J Dent Res. 1979 Apr;58(4):1415-23
PMID: 285112
-
Gap junctional communication during limb cartilage differentiation.
Dev Biol. 1991 Mar;144(1):47-53
PMID: 1995401
-
Membrane modifications in chick osteoclasts revealed by freeze-fracture replicas.
Am J Anat. 1990 Aug;188(4):381-92
PMID: 2392994
-
Osteoblasts and collagen orientation.
Cell Tissue Res. 1975 May 27;159(1):73-80
PMID: 1149092
-
Secretory territories and rate of matrix production of osteoblasts.
Calcif Tissue Res. 1974 May 28;14(4):309-15
PMID: 4843778
-
Prostaglandins change cell shape and increase intercellular gap junctions in osteoblasts cultured from rat fetal calvaria.
J Bone Miner Res. 1986 Jun;1(3):243-9
PMID: 3503541
-
Gap junction distribution in adult mammalian myocardium revealed by an anti-peptide antibody and laser scanning confocal microscopy.
J Cell Sci. 1991 May;99 ( Pt 1):41-55
PMID: 1661743
-
The bone lining cell: a distinct phenotype?
Calcif Tissue Int. 1987 Jul;41(1):1-5
PMID: 3113697
-
Colonization of various natural substrates by osteoblasts in vitro.
Scan Electron Microsc. 1979;(2):529-38
PMID: 524023
-
Bone lining cells: structure and function.
Scanning Microsc. 1989 Sep;3(3):953-60; discussion 960-1
PMID: 2694361
-
Scanning electron microscopic observations of rat tibia using the HCl-collagenase method.
Arch Histol Jpn. 1982 Sep;45(4):399-404
PMID: 6299226
-
A study of human root cementum surfaces as prepared for and examined in the scanning electron microscope.
Z Zellforsch Mikrosk Anat. 1972;130(3):318-37
PMID: 4560319
-
Direct cell-cell communication in the blood-forming system.
Tissue Cell. 1991;23(4):457-70
PMID: 1926137
-
Topological analysis of the major protein in isolated intact rat liver gap junctions and gap junction-derived single membrane structures.
J Biol Chem. 1987 Jun 5;262(16):7751-63
PMID: 3034905
-
Scrape-loading and dye transfer. A rapid and simple technique to study gap junctional intercellular communication.
Exp Cell Res. 1987 Feb;168(2):422-30
PMID: 2433137
-
Characterization of gap junctions between osteoblast-like cells in culture.
Calcif Tissue Int. 1992 Oct;51(4):285-90
PMID: 1330238
-
Cardiac myocyte gap junctions: evidence for a major connexon protein with an apparent relative molecular mass of 70,000.
J Cell Sci. 1990 Aug;96 ( Pt 4):591-604
PMID: 2178163
-
Occurrence of cell junctions and microfilaments in osteoblasts.
Cell Tissue Res. 1975 Jun 13;159(3):413-22
PMID: 1149107
-
Confocal fluorescence microscopy: some applications in bone cell biology.
J Microsc. 1990 May;158(Pt 2):249-59
PMID: 2115085
-
Constants in lamellar bone formation determined by osteoblast kinetics.
Bone. 1985;6(5):361-3
PMID: 4096871
-
The gap junction.
J Cell Sci. 1988 Jan;89 ( Pt 1):1-7
PMID: 2458367
-
Cardiac gap junctions in rat ventricle: localization using site-directed antibodies and laser scanning confocal microscopy.
Cardioscience. 1990 Mar;1(1):75-82
PMID: 1966373
-
Cortical bone repair. The relationship of the lacunar-canalicular system and intercellular gap junctions to the repair process.
J Bone Joint Surg Am. 1988 Aug;70(7):1067-81
PMID: 3042791
-
An ultrastructural study of bone cells: the occurrence of microtubules, microfilaments and tight junctions.
Calcif Tissue Res. 1974;14(1):15-29
PMID: 4820235
-
Morphological evidence of gap junctions between bone cells.
Calcif Tissue Int. 1981;33(5):509-12
PMID: 6797704