Intercellular junctions and the application of microscopical techniques: the cardiac gap junction as a case model

Intercellular junctions are fundamental to the interactions between cells. By means of these junctions, the activities of the individual cells that make up tissues are co‐ordinated, enabling each tissue system to function as an integrated whole. In this review, the work of the authors on one specific type of junction—the cardiac gap junction—is presented as a case model to illustrate how the application of a range of microscopical methods, as part of a multidisciplinary approach, can help extend our understanding of cell junctions and their functions. In the heart, gap junctions form the low‐resistance pathways for rapid impulse conduction and propagation, enabling synchronous stimulation of myocyte contraction. Gap junctions also form pathways for direct intercellular communication, a function of particular importance for morphogenetic signalling during development. The work discussed demonstrates some of the applications of techniques in electron microscopy, immunocytochemistry and confocal scanning laser microscopy to the understanding of the structural basis of the function of gap junctions in the normal adult heart, the developing heart and the diseased heart.

[1]  C. Green,et al.  Evidence for a distinct gap-junctional phenotype in ventricular conduction tissues of the developing and mature avian heart. , 1993, Circulation research.

[2]  K. Willecke,et al.  Molecular cloning and functional expression of mouse connexin40, a second gap junction gene preferentially expressed in lung , 1992, The Journal of cell biology.

[3]  J. Saffitz,et al.  Cardiac myocyte interconnections at gap junctions Role in normal and abnormal electrical conduction. , 1992, Trends in cardiovascular medicine.

[4]  G. Fishman Connexins and the heart. , 1992, Trends in cardiovascular medicine.

[5]  M. Yeager,et al.  Membrane topology and quaternary structure of cardiac gap junction ion channels. , 1992, Journal of molecular biology.

[6]  J. Saffitz,et al.  Cardiac myocytes express multiple gap junction proteins. , 1992, Circulation research.

[7]  J. Revel,et al.  Spatial and temporal patterns of distribution of the gap junction protein connexin43 during mouse gastrulation and organogenesis. , 1992, Development.

[8]  C. Green,et al.  Altered patterns of gap junction distribution in ischemic heart disease. An immunohistochemical study of human myocardium using laser scanning confocal microscopy. , 1991, The American journal of pathology.

[9]  A. Magee,et al.  Transmembrane molecular assemblies regulated by the greater cadherin family. , 1991, Current opinion in cell biology.

[10]  K. Willecke,et al.  The diversity of connexin genes encoding gap junctional proteins. , 1991, European journal of cell biology.

[11]  N. Unwin,et al.  Isolation and purification of gap junction channels , 1991, The Journal of cell biology.

[12]  R. Lal,et al.  Atomic force microscopy and dissection of gap junctions , 1991, Science.

[13]  N. Gilula,et al.  Spatiotemporal expression of three gap junction gene products involved in fetomaternal communication during rat pregnancy. , 1991, Development.

[14]  N. Severs The structure and function of cardiac intercellular junctions , 1991 .

[15]  G. Wheeler,et al.  Desmosomal glycoproteins II and III. Cadherin-like junctional molecules generated by alternative splicing. , 1991, The Journal of biological chemistry.

[16]  W. Nayler,et al.  What constitutes the calcium paradox? , 1991, Journal of molecular and cellular cardiology.

[17]  T. Ruigrok,et al.  The intracellular Na+ concentration prior to Ca2+ repletion has no bearing on the occurrence of the calcium paradox as originally defined. , 1991, Journal of molecular and cellular cardiology.

[18]  M. Suleiman,et al.  The calcium paradox: a role for [Na]i, a cellular or tissue basis, a property unique to the Langendorff perfused heart? A bundle of contradictions! , 1991, Journal of molecular and cellular cardiology.

[19]  W H Lamers,et al.  Spatial distribution of connexin43, the major cardiac gap junction protein, in the developing and adult rat heart. , 1991, Circulation research.

[20]  C. Green,et al.  Gap junction distribution in adult mammalian myocardium revealed by an anti-peptide antibody and laser scanning confocal microscopy. , 1991, Journal of cell science.

[21]  J E Saffitz,et al.  Remodeling of ventricular conduction pathways in healed canine infarct border zones. , 1991, The Journal of clinical investigation.

[22]  E. Hertzberg,et al.  Gap junctions: New tools, new answers, new questions , 1991, Neuron.

[23]  H. Jongsma,et al.  The Cardiac Connection , 1991 .

[24]  J. Revel,et al.  Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes. , 1990, Journal of cell science.

[25]  C. Green,et al.  Cardiac myocyte gap junctions: evidence for a major connexon protein with an apparent relative molecular mass of 70,000. , 1990, Journal of cell science.

[26]  C. Green,et al.  Antibodies to cardiac gap-junctional protein: Biochemical and morphological applications , 1990 .

[27]  D C Spray,et al.  Structure-activity relations of the cardiac gap junction channel. , 1990, The American journal of physiology.

[28]  C. Green,et al.  Cardiac gap junctions in rat ventricle: localization using site-directed antibodies and laser scanning confocal microscopy. , 1990, Cardioscience.

[29]  N. Severs The cardiac gap junction and intercalated disc. , 1990, International journal of cardiology.

[30]  M. Takeichi,et al.  Cadherins: a molecular family important in selective cell-cell adhesion. , 1990, Annual review of biochemistry.

[31]  P. Jones,et al.  Identification of a talin binding site in the cytoskeletal protein vinculin , 1989, The Journal of cell biology.

[32]  J. Saffitz,et al.  Quantitative Analysis of Intercellular Connections by Immunohistochemistry of the Cardiac Gap Junction Protein Connexin43 , 1989, Circulation research.

[33]  N. Severs Gap junction shape and orientation at the cardiac intercalated disk. , 1989, Circulation research.

[34]  D. M. Shotton,et al.  Confocal scanning optical microscopy and its applications for biological specimens , 1989 .

[35]  C. Green,et al.  Fate of Gap Junctions in Isolated Adult Mammalian Cardiomyocytes , 1989, Circulation research.

[36]  R. Lal,et al.  The 43-kD polypeptide of heart gap junctions: immunolocalization, topology, and functional domains , 1989, The Journal of cell biology.

[37]  J E Saffitz,et al.  Distribution and Three‐Dimensional Structure of Intercellular Junctions in Canine Myocardium , 1989, Circulation research.

[38]  D. Paul,et al.  Antisera directed against connexin43 peptides react with a 43-kD protein localized to gap junctions in myocardium and other tissues , 1989, The Journal of cell biology.

[39]  R. Houghten,et al.  Topology of the 32‐kd liver gap junction protein determined by site‐directed antibody localizations. , 1988, The EMBO journal.

[40]  C. Green,et al.  Gap junctions from rabbit heart and dissociated myocytes — Development of detergent—Free isolation methods☆ , 1988 .

[41]  Benjamin Geiger,et al.  Cingulin, a new peripheral component of tight junctions , 1988, Nature.

[42]  C. Green Evidence mounts for the role of gap junctions during development , 1988, BioEssays : news and reviews in molecular, cellular and developmental biology.

[43]  B. Gumbiner,et al.  Structure, biochemistry, and assembly of epithelial tight junctions. , 1987, The American journal of physiology.

[44]  D. Paul,et al.  Connexin43: a protein from rat heart homologous to a gap junction protein from liver , 1987, The Journal of cell biology.

[45]  J. Frank,et al.  Membrane structure in ultrarapidly frozen, unpretreated, freeze-fractured myocardium. , 1987, Circulation research.

[46]  M. Fordham,et al.  An evaluation of confocal versus conventional imaging of biological structures by fluorescence light microscopy , 1987, The Journal of cell biology.

[47]  C. Green,et al.  Topological analysis of the major protein in isolated intact rat liver gap junctions and gap junction-derived single membrane structures. , 1987, The Journal of biological chemistry.

[48]  J D Pitts,et al.  The Gap Junction , 1986, Journal of Cell Science.

[49]  B. Menco A survey of ultra‐rapid cryofixation methods with particular emphasis on applications to freeze‐fracturing, freeze‐etching, and freeze‐substitution , 1986 .

[50]  U. Sleytr,et al.  Low Temperature Methods in Biological Electron Microscopy , 1985 .

[51]  B. Geiger,et al.  Molecular heterogeneity of adherens junctions , 1985, The Journal of cell biology.

[52]  W. Nayler,et al.  Contracture and the calcium paradox. , 1985, Journal of molecular and cellular cardiology.

[53]  C. Manjunath,et al.  Cell biology and protein composition of cardiac gap junctions. , 1985, The American journal of physiology.

[54]  W. J. Larsen,et al.  The dynamic life histories of intercellular membrane junctions , 1985 .

[55]  N. Severs,et al.  Ultrastructure of the sarcolemma and intercalated disc in isolated rat myocytes. , 1985, Basic research in cardiology.

[56]  N. Sperelakis,et al.  Intercalated discs of mammalian heart: a review of structure and function. , 1985, Tissue & cell.

[57]  V. Koteliansky,et al.  The role of actin-binding proteins vinculin, filamin, and fibronectin in intracellular and intercellular linkages in cardiac muscle. , 1985, Advances in myocardiology.

[58]  N. Severs Intercellular junctions and the cardiac intercalated disk. , 1985, Advances in myocardiology.

[59]  C. Green,et al.  Gap junction connexon configuration in rapidly frozen myocardium and isolated intercalated disks , 1984, The Journal of cell biology.

[60]  G. Goings,et al.  Cytoplasmic surface and intramembrane components of rat heart gap junctional proteins. , 1984, The American journal of physiology.

[61]  P. N. Unwin,et al.  Two configurations of a channel-forming membrane protein , 1984, Nature.

[62]  N. Severs,et al.  Isolated calcium-tolerant myocytes and the calcium paradox: an ultrastructural comparison. , 1983, European heart journal.

[63]  P. Pinto da Silva,et al.  In vitro, rapid assembly of gap junctions is induced by cytoskeleton disruptors , 1983, The Journal of cell biology.

[64]  T. Karrison,et al.  Freeze-fractured cardiac gap junctions: structural analysis by three methods. , 1983, The American journal of physiology.

[65]  C. Green,et al.  A simplified method for the rapid isolation of cardiac intercalated discs. , 1983, Tissue & cell.

[66]  N. Severs,et al.  Correlation of ultrastructure and function in calcium-tolerant myocytes isolated from the adult rat heart. , 1982, Journal of ultrastructure research.

[67]  J. Escaig New instruments which facilitate rapid freezing at 83 K and 6 K , 1982 .

[68]  U. Sleytr,et al.  Understanding the artefact problem in freeze‐fracture replication: a review , 1982, Journal of microscopy.

[69]  H. Y. Elder,et al.  Optimum conditions for cryoquenching of small tissue blocks in liquid coolants , 1982, Journal of microscopy.

[70]  T. Reese,et al.  Evidence for the lipidic nature of tight junction strands , 1982, Nature.

[71]  B. Kachar,et al.  On tight-junction structure , 1982, Cell.

[72]  E. Page,et al.  Gap junctional structure in intact and cut sheep cardiac Purkinje fibers: a freeze-fracture study of Ca2+-induced resealing. , 1981, Journal of ultrastructure research.

[73]  T. Pexieder Mechanisms of Cardiac Morphogenesis and Teratogenesis , 1981 .

[74]  G. Zampighi,et al.  Structure of the junction between communicating cells , 1980, Nature.

[75]  D. Shotton Quick-freezing — the new frontier in freeze-fracture , 1980, Nature.

[76]  C. Peracchia Structural correlates of gap junction permeation. , 1980, International review of cytology.

[77]  J. Rash,et al.  Freeze Fracture: Methods, Artifacts, and Interpretations , 1979 .

[78]  K. Baldwin Cardiac gap junction configuration after an uncoupling treatment as a function of time , 1979, The Journal of cell biology.

[79]  D. Gros,et al.  Formation and growth of gap junctions in mouse myocardium during ontogenesis: quantitative data and their implications on the development of intercellular communication. , 1979, Journal of molecular and cellular cardiology.

[80]  M. Dennis,et al.  Synaptic vesicle exocytosis captured by quick freezing and correlated with quantal transmitter release , 1979, The Journal of cell biology.

[81]  C. Halverson,et al.  Ultrastructural modifications of nexuses (gap junctions) during early myocardial ischemia. , 1978, Journal of molecular and cellular cardiology.

[82]  L. Makowski,et al.  Gap junction structures: Analysis of the x-ray diffraction data , 1977, The Journal of cell biology.

[83]  G. P. Dempsey,et al.  A copper block method for freezing non‐cryoprotected tissue to produce ice‐crystal‐free regions for electron microscopy , 1976, Journal of microscopy.

[84]  J. Revel,et al.  HEXAGONAL ARRAY OF SUBUNITS IN INTERCELLULAR JUNCTIONS OF THE MOUSE HEART AND LIVER , 1967, The Journal of cell biology.

[85]  W. C. Hülsmann,et al.  Paradoxical Influence of Calcium Ions on the Permeability of the Cell Membranes of the Isolated Rat Heart , 1966, Nature.

[86]  L. Barr,et al.  Propagation of Action Potentials and the Structure of the Nexus in Cardiac Muscle , 1965, The Journal of general physiology.