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Three-dimensional reconstruction of the intercalated disc including the intercellular junctions by applying volume scanning electron microscopy

(2018) HISTOCHEMISTRY AND CELL BIOLOGY. 149(5). p.479-490
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Abstract
The intercalated disc (ID) contains different kinds of intercellular junctions: gap junctions (GJs), desmosomes and areae compositae, essential for adhesion and communication between adjacent cardiomyocytes. The junctions can be identified based on their morphology when imaged using transmission electron microscopy (TEM), however, only with very limited information in the z-dimension. The application of volume EM techniques can give insight into the three-dimensional (3-D) organization of complex biological structures. In this study, we generated 3-D datasets using serial block-face scanning electron microscopy (SBF-SEM) and focused ion beam SEM (FIB-SEM), the latter resulting in datasets with 5 nm isotropic voxels. We visualized cardiomyocytes in murine ventricular heart tissue and, for the first time, we could three-dimensionally reconstruct the ID including desmosomes and GJs with 5 nm precision in a large volume. Results show in three dimensions a highly folded structure of the ID, with the presence of GJs and desmosomes in both plicae and interplicae regions. We observed close contact of GJs with mitochondria and a variable spatial distribution of the junctions. Based on measurements of the shape of the intercellular junctions in 3-D, it is seen that GJs and desmosomes vary in size, depending on the region within the ID. This demonstrates that volume EM is essential to visualize morphological changes and its potential to quantitatively determine structural changes between normal and pathological conditions, e.g., cardiomyopathies.
Keywords
Cardiomyocytes, Cardiac intercalated disc, Desmosome, Gap junction, Volume electron microscopy, ALPHA-T-CATENIN, HEART-MUSCLE CELLS, GAP-JUNCTIONS, ADHERING JUNCTIONS, AREA-COMPOSITA, DILATED CARDIOMYOPATHY, TRANSITIONAL JUNCTION, ADHERENS JUNCTION, BETA-CATENIN, CONNEXIN43

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Citation

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Chicago
Vanslembrouck, Bieke, Anna Kremer, Benjamin Pavie, Frans Van Roy, Saskia Lippens, and Jolanda van Hengel. 2018. “Three-dimensional Reconstruction of the Intercalated Disc Including the Intercellular Junctions by Applying Volume Scanning Electron Microscopy.” Histochemistry and Cell Biology 149 (5): 479–490.
APA
Vanslembrouck, B., Kremer, A., Pavie, B., Van Roy, F., Lippens, S., & van Hengel, J. (2018). Three-dimensional reconstruction of the intercalated disc including the intercellular junctions by applying volume scanning electron microscopy. HISTOCHEMISTRY AND CELL BIOLOGY, 149(5), 479–490.
Vancouver
1.
Vanslembrouck B, Kremer A, Pavie B, Van Roy F, Lippens S, van Hengel J. Three-dimensional reconstruction of the intercalated disc including the intercellular junctions by applying volume scanning electron microscopy. HISTOCHEMISTRY AND CELL BIOLOGY. 2018;149(5):479–90.
MLA
Vanslembrouck, Bieke, Anna Kremer, Benjamin Pavie, et al. “Three-dimensional Reconstruction of the Intercalated Disc Including the Intercellular Junctions by Applying Volume Scanning Electron Microscopy.” HISTOCHEMISTRY AND CELL BIOLOGY 149.5 (2018): 479–490. Print.
@article{8553333,
  abstract     = {The intercalated disc (ID) contains different kinds of intercellular junctions: gap junctions (GJs), desmosomes and areae compositae, essential for adhesion and communication between adjacent cardiomyocytes. The junctions can be identified based on their morphology when imaged using transmission electron microscopy (TEM), however, only with very limited information in the z-dimension. The application of volume EM techniques can give insight into the three-dimensional (3-D) organization of complex biological structures. In this study, we generated 3-D datasets using serial block-face scanning electron microscopy (SBF-SEM) and focused ion beam SEM (FIB-SEM), the latter resulting in datasets with 5 nm isotropic voxels. We visualized cardiomyocytes in murine ventricular heart tissue and, for the first time, we could three-dimensionally reconstruct the ID including desmosomes and GJs with 5 nm precision in a large volume. Results show in three dimensions a highly folded structure of the ID, with the presence of GJs and desmosomes in both plicae and interplicae regions. We observed close contact of GJs with mitochondria and a variable spatial distribution of the junctions. Based on measurements of the shape of the intercellular junctions in 3-D, it is seen that GJs and desmosomes vary in size, depending on the region within the ID. This demonstrates that volume EM is essential to visualize morphological changes and its potential to quantitatively determine structural changes between normal and pathological conditions, e.g., cardiomyopathies.},
  author       = {Vanslembrouck, Bieke and Kremer, Anna and Pavie, Benjamin and Van Roy, Frans and Lippens, Saskia and van Hengel, Jolanda},
  issn         = {0948-6143},
  journal      = {HISTOCHEMISTRY AND CELL BIOLOGY},
  language     = {eng},
  number       = {5},
  pages        = {479--490},
  title        = {Three-dimensional reconstruction of the intercalated disc including the intercellular junctions by applying volume scanning electron microscopy},
  url          = {http://dx.doi.org/10.1007/s00418-018-1657-x},
  volume       = {149},
  year         = {2018},
}

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