Williams, Marion Patrick Ivey and Rigon, Matteo and Straka, Tatjana and Hörner, Sarah Janice and Thiel, Manfred and Gretz, Norbert and Hafner, Mathias and Reischl, Markus and Rudolf, Rüdiger (2019) A Novel Optical Tissue Clearing Protocol for Mouse Skeletal Muscle to Visualize Endplates in Their Tissue Context. Frontiers in Cellular Neuroscience, 13. ISSN 1662-5102
pubmed-zip/versions/1/package-entries/fncel-13-00049/fncel-13-00049.pdf - Published Version
Download (16MB)
Abstract
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.
Item Type: | Article |
---|---|
Subjects: | STM Digital Press > Medical Science |
Depositing User: | Unnamed user with email support@stmdigipress.com |
Date Deposited: | 29 May 2023 05:52 |
Last Modified: | 02 Sep 2024 12:40 |
URI: | http://publications.articalerewriter.com/id/eprint/951 |