Journal of Cell Science, Vol 99, Issue 1 41-55, Copyright © 1991 by Company of Biologists

JOURNAL ARTICLES

Gap junction distribution in adult mammalian myocardium revealed by an anti-peptide antibody and laser scanning confocal microscopy

RG Gourdie, CR Green and NJ Severs
Department of Anatomy and Developmental Biology, University College London, UK.

A polyclonal antiserum, raised against a synthetic peptide matching part of the sequence of connexin43 (a rat cardiac gap-junctional protein), was used in combination with laser scanning confocal microscopy to investigate gap junction distribution in cardiac tissues from a range of mammalian species. Comparison of the localised punctate staining patterns obtained in ventricular tissue with the distribution of intercalated disks as viewed by conventional light microscopy and electron microscopy, and with the staining observed by standard light-microscope immunofluorescence using the same anti-serum, demonstrated highly specific labelling of clearly resolved individual gap junctions. Laser scanning confocal microscopy of ventricular myocardium showed the immunostained gap junctions to be confined to well-defined intercalated disks bisecting the long axis of the muscle fibre, whereas in the atrial myocardium, gap junctions were commonly distributed widely over the lateral surfaces of the myocyte body. Rat atrial gap junctions were significantly larger (as measured by the longest axial lengths of fluorescent spots), and showed a narrower spread of sizes, than their counterparts in the ventricle. Ventricular myocardium from six mammalian species including man gave similar immunostaining patterns, indicating conservation both of the epitope(s) detected by the antiserum, and of the general organisation of the cell-to-cell pathways for electrical propagation, in the mammalian heart. Optical section series obtained by laser scanning confocal microscopy permitted the quantification and mapping of the three-dimensional distribution of gap junctions in ventricular intercalated disks with high clarity over substantial specimen depths. A consistent feature of gap junction organisation within disks of ventricular myocardium in all species studied was the presence of a conspicuous ring of large gap junctions around the periphery of the disk. Immunostained gap junctions lying within the interior zone delineated by the peripheral junctions generally occurred at lower numerical densities and were significantly smaller. In all species, less than 3% of all immunolabelled gap junctions measured were greater than 2 microns in maximal length, though a small proportion (0.06%) exceeded 4 microns. The numerical density of immunolabelled gap junctions in the disk was similar between species; however, within species there was a significant decrease in numerical density with increasing disk size. The new features of intercalated disk structure revealed in this study may have an important part to play in the intercellular communication and electrical propagation properties of the mammalian heart.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				N. J. Severs The Carboxy Terminal Domain of Connexin43: From Molecular Regulation of the Gap Junction Channel to Supramolecular Organization of the Intercalated Disk Circ. Res., December 7, 2007; 101(12): 1213 - 1215. [Full Text] [PDF]

				K. W. Hewett, L. W. Norman, D. Sedmera, R. J. Barker, C. Justus, J. Zhang, S. W. Kubalak, and R. G. Gourdie Knockout of the neural and heart expressed gene HF-1b results in apical deficits of ventricular structure and activation Cardiovasc Res, August 15, 2005; 67(3): 548 - 560. [Abstract] [Full Text] [PDF]

				N. Zeevi-Levin, Y. D. Barac, Y. Reisner, I. Reiter, G. Yaniv, G. Meiry, Z. Abassi, S. Kostin, J. Schaper, M. R. Rosen, et al. Gap junctional remodeling by hypoxia in cultured neonatal rat ventricular myocytes Cardiovasc Res, April 1, 2005; 66(1): 64 - 73. [Abstract] [Full Text] [PDF]

				P. Saitongdee, D. L. Becker, P. Milner, G. E. Knight, and G. Burnstock Levels of Gap Junction Proteins in Coronary Arterioles and Aorta of Hamsters Exposed to the Cold and During Hibernation and Arousal J. Histochem. Cytochem., May 1, 2004; 52(5): 603 - 616. [Abstract] [Full Text] [PDF]

				N. J. Severs, S. R. Coppen, E. Dupont, H.-I Yeh, Y.-S. Ko, and T. Matsushita Gap junction alterations in human cardiac disease Cardiovasc Res, May 1, 2004; 62(2): 368 - 377. [Abstract] [Full Text] [PDF]

				S. Kostin, S. Dammer, S. Hein, W. P Klovekorn, E. P Bauer, and J. Schaper Connexin 43 expression and distribution in compensated and decompensated cardiac hypertrophy in patients with aortic stenosis Cardiovasc Res, May 1, 2004; 62(2): 426 - 436. [Abstract] [Full Text] [PDF]

				S. P. Thomas, J. P. Kucera, L. Bircher-Lehmann, Y. Rudy, J. E. Saffitz, and A. G. Kleber Impulse Propagation in Synthetic Strands of Neonatal Cardiac Myocytes With Genetically Reduced Levels of Connexin43 Circ. Res., June 13, 2003; 92(11): 1209 - 1216. [Abstract] [Full Text] [PDF]

				D. E. Gutstein, F.-y. Liu, M. B. Meyers, A. Choo, and G. I. Fishman The organization of adherens junctions and desmosomes at the cardiac intercalated disc is independent of gap junctions J. Cell Sci., March 1, 2003; 116(5): 875 - 885. [Abstract] [Full Text] [PDF]

				R. J. Barker and R. G. Gourdie JNK Bond Regulation: Why Do Mammalian Hearts Invest in Connexin43? Circ. Res., October 4, 2002; 91(7): 556 - 558. [Full Text] [PDF]

				R. J. Barker, R. L. Price, and R. G. Gourdie Increased Association of ZO-1 With Connexin43 During Remodeling of Cardiac Gap Junctions Circ. Res., February 22, 2002; 90(3): 317 - 324. [Abstract] [Full Text] [PDF]

				W. Huang, M. P Kingsbury, M. A Turner, J.L. Donnelly, N. A Flores, and D. J Sheridan Capillary filtration is reduced in lungs adapted to chronic heart failure: morphological and haemodynamic correlates Cardiovasc Res, January 1, 2001; 49(1): 207 - 217. [Abstract] [Full Text] [PDF]

				S. P. Thomas, L. Bircher-Lehmann, S. A. Thomas, J. Zhuang, J. E. Saffitz, and A. G. Kleber Synthetic Strands of Neonatal Mouse Cardiac Myocytes : Structural and Electrophysiological Properties Circ. Res., September 15, 2000; 87(6): 467 - 473. [Abstract] [Full Text] [PDF]

				M.R. Boyett, H. Honjo, and I. Kodama The sinoatrial node, a heterogeneous pacemaker structure Cardiovasc Res, September 1, 2000; 47(4): 658 - 687. [Abstract] [Full Text] [PDF]

				P. Saitongdee, P. Milner, D. L Becker, G. E Knight, and G. Burnstock Increased connexin43 gap junction protein in hamster cardiomyocytes during cold acclimatization and hibernation Cardiovasc Res, July 1, 2000; 47(1): 108 - 115. [Abstract] [Full Text] [PDF]

				M. S. Spach, J. F. Heidlage, P. C. Dolber, and R. C. Barr Electrophysiological Effects of Remodeling Cardiac Gap Junctions and Cell Size : Experimental and Model Studies of Normal Cardiac Growth Circ. Res., February 18, 2000; 86(3): 302 - 311. [Abstract] [Full Text] [PDF]

				M. S. Spach, J. F. Heidlage, P. C. Dolber, and R. C. Barr Extracellular Discontinuities in Cardiac Muscle : Evidence for Capillary Effects on the Action Potential Foot Circ. Res., November 30, 1998; 83(11): 1144 - 1164. [Abstract] [Full Text] [PDF]

				S. Verheule, M. J. A. van Kempen, P. H. J. A. t. Welscher, B. R. Kwak, and H. J. Jongsma Characterization of Gap Junction Channels in Adult Rabbit Atrial and Ventricular Myocardium Circ. Res., May 19, 1997; 80(5): 673 - 681. [Abstract] [Full Text]

				B. Nadarajah, A. M. Jones, W. H. Evans, and J. G. Parnavelas Differential Expression of Connexins during Neocortical Development and Neuronal Circuit Formation J. Neurosci., May 1, 1997; 17(9): 3096 - 3111. [Abstract] [Full Text] [PDF]

				N. S. Peters, J. Coromilas, N. J. Severs, and A. L. Wit Disturbed Connexin43 Gap Junction Distribution Correlates With the Location of Reentrant Circuits in the Epicardial Border Zone of Healing Canine Infarcts That Cause Ventricular Tachycardia Circulation, February 18, 1997; 95(4): 988 - 996. [Abstract] [Full Text]

				B. D. Angst, L. U.R. Khan, N. J. Severs, K. Whitely, S. Rothery, R. P. Thompson, A. I. Magee, and R. G. Gourdie Dissociated Spatial Patterning of Gap Junctions and Cell Adhesion Junctions During Postnatal Differentiation of Ventricular Myocardium Circ. Res., January 1, 1997; 80(1): 88 - 94. [Abstract] [Full Text]

				J. P. Blackburn, N. S. Peters, H.-I. Yeh, S. Rothery, C. R. Green, and N. J. Severs Upregulation of Connexin43 Gap Junctions During Early Stages of Human Coronary Atherosclerosis Arterioscler Thromb Vasc Biol, August 1, 1995; 15(8): 1219 - 1228. [Abstract] [Full Text]

				M. S. Spach and J. F. Heidlage The Stochastic Nature of Cardiac Propagation at a Microscopic Level : Electrical Description of Myocardial Architecture and Its Application to Conduction Circ. Res., March 1, 1995; 76(3): 366 - 380. [Abstract] [Full Text]

				R. Gourdie, T Mima, R. Thompson, and T Mikawa Terminal diversification of the myocyte lineage generates Purkinje fibers of the cardiac conduction system Development, January 5, 1995; 121(5): 1423 - 1431. [Abstract] [PDF]

				M Rosendaal, C. Green, A Rahman, and D Morgan Up-regulation of the connexin43+ gap junction network in haemopoietic tissue before the growth of stem cells J. Cell Sci., January 1, 1994; 107(1): 29 - 37. [Abstract] [PDF]

				R. Gourdie, N. Severs, C. Green, S Rothery, P Germroth, and R. Thompson The spatial distribution and relative abundance of gap-junctional connexin40 and connexin43 correlate to functional properties of components of the cardiac atrioventricular conduction system J. Cell Sci., January 8, 1993; 105(4): 985 - 991. [Abstract] [PDF]

				R. J. Barker, R. L. Price, and R. G. Gourdie Increased Association of ZO-1 With Connexin43 During Remodeling of Cardiac Gap Junctions Circ. Res., February 22, 2002; 90(3): 317 - 324. [Abstract] [Full Text] [PDF]

				S. Kostin and J. Schaper Tissue-Specific Patterns of Gap Junctions in Adult Rat Atrial and Ventricular Cardiomyocytes In Vivo and In Vitro Circ. Res., May 11, 2001; 88(9): 933 - 939. [Abstract] [Full Text] [PDF]