A Novel Optical Tissue Clearing Protocol for Mouse Skeletal Muscle to Visualize Endplates in Their Tissue Context
Neuromuscular junctions (NMJs) mediate skeletal muscle contractions and play an important role in several neuromuscular disorders when their morphology and function are compromised. However, due to their small size and sparse distribution throughout the comparatively large, inherently opaque muscle...
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doaj-c33ebd6958c84e2b8b14bf79b5c2d9dd2020-11-24T21:17:01ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022019-02-011310.3389/fncel.2019.00049430452A Novel Optical Tissue Clearing Protocol for Mouse Skeletal Muscle to Visualize Endplates in Their Tissue ContextMarion Patrick Ivey Williams0Matteo Rigon1Tatjana Straka2Tatjana Straka3Sarah Janice Hörner4Sarah Janice Hörner5Manfred Thiel6Norbert Gretz7Norbert Gretz8Mathias Hafner9Mathias Hafner10Markus Reischl11Rüdiger Rudolf12Rüdiger Rudolf13Rüdiger Rudolf14Rüdiger Rudolf15Institute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, GermanyInstitute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, GermanyInstitute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, GermanyInterdisciplinary Center for Neurosciences, Heidelberg University, Heidelberg, GermanyInstitute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, GermanyInterdisciplinary Center for Neurosciences, Heidelberg University, Heidelberg, GermanyDepartment of Anesthesiology and Surgical Intensive Care Medicine, Medical Faculty Mannheim, Heidelberg University, Mannheim, GermanyMedical Faculty Mannheim, Medical Research Center, Heidelberg University, Mannheim, GermanyMedical Faculty Mannheim, Institute of Medical Technology, Mannheim University of Applied Sciences, Mannheim, GermanyInstitute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, GermanyMedical Faculty Mannheim, Institute of Medical Technology, Mannheim University of Applied Sciences, Mannheim, GermanyInstitute for Automation and Applied Informatics, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, GermanyInstitute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, GermanyInterdisciplinary Center for Neurosciences, Heidelberg University, Heidelberg, GermanyMedical Faculty Mannheim, Institute of Medical Technology, Mannheim University of Applied Sciences, Mannheim, GermanyInstitute of Toxicology and Genetics, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, GermanyNeuromuscular junctions (NMJs) mediate skeletal muscle contractions and play an important role in several neuromuscular disorders when their morphology and function are compromised. However, due to their small size and sparse distribution throughout the comparatively large, inherently opaque muscle tissue the analysis of NMJ morphology has been limited to teased fiber preparations, longitudinal muscle sections, and flat muscles. Consequently, whole mount analyses of NMJ morphology, numbers, their distribution, and assignment to a given muscle fiber have also been impossible to determine in muscle types that are frequently used in experimental paradigms. This impossibility is exacerbated by the lack of optical tissue clearing techniques that are compatible with clear and persistent NMJ stains. Here, we present MYOCLEAR, a novel and highly reproducible muscle tissue clearing protocol. Based on hydrogel-based tissue clearing methods, this protocol permits the labeling and detection of all NMJs in adult hindleg extensor digitorum longus muscles from wildtype and diseased mice. The method is also applicable to adult mouse diaphragm muscles and can be used for different staining agents, including toxins, lectins, antibodies, and nuclear dyes. It will be useful in understanding the distribution, morphological features, and muscle tissue context of NMJs in hindleg muscle whole mounts for biomedical and basic research.https://www.frontiersin.org/article/10.3389/fncel.2019.00049/fullacetylcholine receptorhydrogel embeddingNMJoptical tissue clearingskeletal muscle |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Marion Patrick Ivey Williams Matteo Rigon Tatjana Straka Tatjana Straka Sarah Janice Hörner Sarah Janice Hörner Manfred Thiel Norbert Gretz Norbert Gretz Mathias Hafner Mathias Hafner Markus Reischl Rüdiger Rudolf Rüdiger Rudolf Rüdiger Rudolf Rüdiger Rudolf |
spellingShingle |
Marion Patrick Ivey Williams Matteo Rigon Tatjana Straka Tatjana Straka Sarah Janice Hörner Sarah Janice Hörner Manfred Thiel Norbert Gretz Norbert Gretz Mathias Hafner Mathias Hafner Markus Reischl Rüdiger Rudolf Rüdiger Rudolf Rüdiger Rudolf Rüdiger Rudolf A Novel Optical Tissue Clearing Protocol for Mouse Skeletal Muscle to Visualize Endplates in Their Tissue Context Frontiers in Cellular Neuroscience acetylcholine receptor hydrogel embedding NMJ optical tissue clearing skeletal muscle |
author_facet |
Marion Patrick Ivey Williams Matteo Rigon Tatjana Straka Tatjana Straka Sarah Janice Hörner Sarah Janice Hörner Manfred Thiel Norbert Gretz Norbert Gretz Mathias Hafner Mathias Hafner Markus Reischl Rüdiger Rudolf Rüdiger Rudolf Rüdiger Rudolf Rüdiger Rudolf |
author_sort |
Marion Patrick Ivey Williams |
title |
A Novel Optical Tissue Clearing Protocol for Mouse Skeletal Muscle to Visualize Endplates in Their Tissue Context |
title_short |
A Novel Optical Tissue Clearing Protocol for Mouse Skeletal Muscle to Visualize Endplates in Their Tissue Context |
title_full |
A Novel Optical Tissue Clearing Protocol for Mouse Skeletal Muscle to Visualize Endplates in Their Tissue Context |
title_fullStr |
A Novel Optical Tissue Clearing Protocol for Mouse Skeletal Muscle to Visualize Endplates in Their Tissue Context |
title_full_unstemmed |
A Novel Optical Tissue Clearing Protocol for Mouse Skeletal Muscle to Visualize Endplates in Their Tissue Context |
title_sort |
novel optical tissue clearing protocol for mouse skeletal muscle to visualize endplates in their tissue context |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Cellular Neuroscience |
issn |
1662-5102 |
publishDate |
2019-02-01 |
description |
Neuromuscular junctions (NMJs) mediate skeletal muscle contractions and play an important role in several neuromuscular disorders when their morphology and function are compromised. However, due to their small size and sparse distribution throughout the comparatively large, inherently opaque muscle tissue the analysis of NMJ morphology has been limited to teased fiber preparations, longitudinal muscle sections, and flat muscles. Consequently, whole mount analyses of NMJ morphology, numbers, their distribution, and assignment to a given muscle fiber have also been impossible to determine in muscle types that are frequently used in experimental paradigms. This impossibility is exacerbated by the lack of optical tissue clearing techniques that are compatible with clear and persistent NMJ stains. Here, we present MYOCLEAR, a novel and highly reproducible muscle tissue clearing protocol. Based on hydrogel-based tissue clearing methods, this protocol permits the labeling and detection of all NMJs in adult hindleg extensor digitorum longus muscles from wildtype and diseased mice. The method is also applicable to adult mouse diaphragm muscles and can be used for different staining agents, including toxins, lectins, antibodies, and nuclear dyes. It will be useful in understanding the distribution, morphological features, and muscle tissue context of NMJs in hindleg muscle whole mounts for biomedical and basic research. |
topic |
acetylcholine receptor hydrogel embedding NMJ optical tissue clearing skeletal muscle |
url |
https://www.frontiersin.org/article/10.3389/fncel.2019.00049/full |
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