A new episcopic method for rapid 3-D reconstruction: applications in anatomy and embryology

Weninger, Wolfgang J.; Meng, Stefan; Streicher, Johannes; Müller, Gerd B.

doi:10.1007/s004290050144

Cited by 73 publications

(61 citation statements)

References 27 publications

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“…High-resolution episcopic images of mouse hearts at E14.5, E16.5, and E 18.5 were obtained as previously described (58). We used the episcopic procedures for sectioning and image capture using a previously described arrangement of microtome and magnification optic (59).…”

Section: Methodsmentioning

confidence: 99%

iASPP, a previously unidentified regulator of desmosomes, prevents arrhythmogenic right ventricular cardiomyopathy (ARVC)-induced sudden death

Notari

Sutendra

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Desmosomes are anchoring junctions that exist in cells that endure physical stress such as cardiac myocytes. The importance of desmosomes in maintaining the homeostasis of the myocardium is underscored by frequent mutations of desmosome components found in human patients and animal models. Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a phenotype caused by mutations in desmosomal components in ∼50% of patients, however, the causes in the remaining 50% of patients still remain unknown. A deficiency of inhibitor of apoptosis-stimulating protein of p53 (iASPP), an evolutionarily conserved inhibitor of p53, caused by spontaneous mutation recently has been associated with a lethal autosomal recessive cardiomyopathy in Poll Hereford calves and Wa3 mice. However, the molecular mechanisms that mediate this putative function of iASPP are completely unknown. Here, we show that iASPP is expressed at intercalated discs in human and mouse postmitotic cardiomyocytes. iASPP interacts with desmoplakin and desmin in cardiomyocytes to maintain the integrity of desmosomes and intermediate filament networks in vitro and in vivo. iASPP deficiency specifically induces right ventricular dilatation in mouse embryos at embryonic day 16.5. iASPPdeficient mice with exon 8 deletion (Ppp1r13l Δ8/Δ8 ) die of sudden cardiac death, displaying features of ARVC. Intercalated discs in cardiomyocytes from four of six human ARVC cases show reduced or loss of iASPP. ARVC-derived desmoplakin mutants DSP-1-V30M and DSP-1-S299R exhibit weaker binding to iASPP. These data demonstrate that by interacting with desmoplakin and desmin, iASPP is an important regulator of desmosomal function both in vitro and in vivo. This newly identified property of iASPP may provide new molecular insight into the pathogenesis of ARVC.ARVC | sudden death | desmosome | iASPP | cell-cell junctions

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Section: Methodsmentioning

confidence: 99%

iASPP, a previously unidentified regulator of desmosomes, prevents arrhythmogenic right ventricular cardiomyopathy (ARVC)-induced sudden death

Notari

Sutendra

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…This includes methods such as "fast 3D serial reconstruction" [39], "Epi-3D" [40], "episcopic fluorescence image capturing" [41], "surface imaging microscopy" [42,43], "high resolution episcopic microscopy (HREM)" [44], and serial blockface scanning electron microscopy (SBF-SEM) [45]. The term "surface imaging microscopy" was used twice.…”

Section: Episcopic 3d Imaging Methods -Definitionmentioning

confidence: 99%

“…"Epi-3D" [40] is designed and was applied for analyzing embryo morphology and the topology of neonate and adult organs respectively [49][50][51][52]. After fixation, the specimens become dehydrated and simultaneously pre-stained with lead acetate, before they are embedded in embedding media on wax basis.…”

Section: Episcopic 3d Imaging Methods -Technical Detailsmentioning

confidence: 99%

Episcopic 3D Imaging Methods: Tools for Researching Gene Function

2013

Advances in Genome Science: Changing Views on Living Organisms

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This work aims at describing episcopic 3D imaging methods and at discussing how these methods can contribute to researching the genetic mechanisms driving embryogenesis and tissue remodelling, and the genesis of pathologies. Several episcopic 3D imaging methods exist. The most advanced are capable of generating high-resolution volume data (voxel sizes from 0.5x0.5x1 m upwards) of small to large embryos of model organisms and tissue samples. Beside anatomy and tissue architecture, gene expression and gene product patterns can be three dimensionally analyzed in their precise anatomical and histological context with the aid of whole mount in situ hybridization or whole mount immunohistochemical staining techniques. Episcopic 3D imaging techniques were and are employed for analyzing the precise morphological phenotype of experimentally malformed, randomly produced, or genetically engineered embryos of biomedical model organisms. It has been shown that episcopic 3D imaging also fits for describing the spatial distribution of genes and gene products during embryogenesis, and that it can be used for analyzing tissue samples of adult model animals and humans. The latter offers the possibility to use episcopic 3D imaging techniques for researching the causality and treatment of pathologies or for staging cancer. Such applications, however, are not yet routine and currently only preliminary results are available. We conclude that, although episcopic 3D imaging is in its very beginnings, it represents an upcoming methodology, which in short terms will become an indispensable tool for researching the genetic regulation of embryo development as well as the genesis of malformations and diseases.

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“…It is however sufficient for 3D analysis of the expression patterns of multiple, selectively labelled (trans-)genes in small parts of embryos [106,107], or the morphology of cells in the walls of the heart and blood vessels. (not all) [108][109][110][111][112]114] Although the table lists the most promising "in vivo techniques", none of them currently enables the generation of useful volume data of the heart and great blood vessels of unborn mice. Note that each of the listed post mortem techniques has its specific advantages and disadvantages.…”

Section: D Confocal Imagingmentioning

confidence: 99%

Three-Dimensional (3D) Visualisation of the Cardiovascular System of Mouse Embryos and Fetus

Weninger¹,

Geyer²

2009

TOCARIJ

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Abstract:The mouse is the most appropriate biomedical model organism for researching the mechanistic function of genetic factors in normal embryogenesis and in the genesis of cardiovascular malformations. Three-dimensional (3D) visualisation of the developing organs of wild type and genetically modified mouse embryos is essential for such research. This paper aims at discussing imaging methods that permit the generation of digital volume data sets, which fit for the virtual 3D visualisation and 3D analysis of the cardiovascular system of mouse embryos and mouse fetus. To cover the spectrum of imaging methods comprehensively, we will start with a short overview about early 3D-reconstruction techniques. This will be followed by a brief discussion why in vivo imaging techniques, such as ultrasound (US) or optical coherence tomography (OCT) do not fit for constructing volume data sets that permit virtual 3D visualisation of the cardiovascular system of mouse embryos/fetus. We will then briefly introduce techniques, which permit 3D analysis of the cardiovascular system but do not fit for creating digital volume data (corrosion casts, scanning electron microscopy). Finally, we will focus on describing the advantages and disadvantages, the spectrum of application and the limitations of important modern digital volume data generation methods, such as micro-computed tomography ( CT), micro-magnetic resonance imaging ( MRI), optical projection tomography (OPT), confocal microscopy, histological section based volume data generation methods, and 3D episcopic imaging methods.

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A new episcopic method for rapid 3-D reconstruction: applications in anatomy and embryology

Cited by 73 publications

References 27 publications

iASPP, a previously unidentified regulator of desmosomes, prevents arrhythmogenic right ventricular cardiomyopathy (ARVC)-induced sudden death

iASPP, a previously unidentified regulator of desmosomes, prevents arrhythmogenic right ventricular cardiomyopathy (ARVC)-induced sudden death

Episcopic 3D Imaging Methods: Tools for Researching Gene Function

Three-Dimensional (3D) Visualisation of the Cardiovascular System of Mouse Embryos and Fetus

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