Cardiac Myocyte Diversity and a Fibroblast Network in the Junctional Region of the Zebrafish Heart Revealed by Transmission and Serial Block-Face Scanning Electron Microscopy
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KAUST DepartmentImaging and Characterization Core Lab
Advanced Nanofabrication, Imaging and Characterization Core Lab
Permanent link to this recordhttp://hdl.handle.net/10754/325323
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AbstractThe zebrafish has emerged as an important model of heart development and regeneration. While the structural characteristics of the developing and adult zebrafish ventricle have been previously studied, little attention has been paid to the nature of the interface between the compact and spongy myocardium. Here we describe how these two distinct layers are structurally and functionally integrated. We demonstrate by transmission electron microscopy that this interface is complex and composed primarily of a junctional region occupied by collagen, as well as a population of fibroblasts that form a highly complex network. We also describe a continuum of uniquely flattened transitional cardiac myocytes that form a circumferential plate upon which the radially-oriented luminal trabeculae are anchored. In addition, we have uncovered within the transitional ring a subpopulation of markedly electron dense cardiac myocytes. At discrete intervals the transitional cardiac myocytes form contact bridges across the junctional space that are stabilized through localized desmosomes and fascia adherentes junctions with adjacent compact cardiac myocytes. Finally using serial block-face scanning electron microscopy, segmentation and volume reconstruction, we confirm the three-dimensional nature of the junctional region as well as the presence of the sheet-like fibroblast network. These ultrastructural studies demonstrate the previously unrecognized complexity with which the compact and spongy layers are structurally integrated, and provide a new basis for understanding development and regeneration in the zebrafish heart. © 2013 Lafontant et al.
CitationLafontant PJ, Behzad AR, Brown E, Landry P, Hu N, et al. (2013) Cardiac Myocyte Diversity and a Fibroblast Network in the Junctional Region of the Zebrafish Heart Revealed by Transmission and Serial Block-Face Scanning Electron Microscopy. PLoS ONE 8: e72388. doi:10.1371/journal.pone.0072388.
PublisherPublic Library of Science (PLoS)
PubMed Central IDPMC3751930
- The intercellular organization of the two muscular systems in the adult salmonid heart, the compact and the spongy myocardium.
- Authors: Pieperhoff S, Bennett W, Farrell AP
- Issue date: 2009 Nov
- Serial block face scanning electron microscopy for the study of cardiac muscle ultrastructure at nanoscale resolutions.
- Authors: Pinali C, Kitmitto A
- Issue date: 2014 Nov
- High resolution scanning electron microscopy of the intracellular surface of intercalated disks in human heart.
- Authors: Tandler B, Riva L, Loy F, Conti G, Isola R
- Issue date: 2006 Dec
- Cytoarchitecture and intercalated disks of the working myocardium and the conduction system in the mammalian heart.
- Authors: Shimada T, Kawazato H, Yasuda A, Ono N, Sueda K
- Issue date: 2004 Oct
- Ultrastructure of the sarcoplasmic reticulum in cardiac myocytes from Pacific bluefin tuna.
- Authors: Di Maio A, Block BA
- Issue date: 2008 Oct
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