Immunohistochemical Detection of Motor Endplates in the Long-Term Denervated Muscle

Mu, Liancai; Chen, Jingming; Li, Jing; Nyirenda, Themba; Fowkes, Mary; Sobotka, Stanislaw

doi:10.1055/s-0038-1627463

Cited by 6 publications

(1 citation statement)

References 71 publications

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“…There are several approaches to obtain the structures of MEPs. Some researchers use traditional immunohistochemical techniques 12-14, but only structural information of extremely thin muscle tissues, such as the diaphragm and the intercostal muscle, can be obtained; other researchers use acetylcholinesterase whole-muscle staining to observe the structure of MEP on the surface of a skeletal muscle 4, 13, but this method cannot provide the distribution information inside the skeletal muscle. There is also a method that combines traditional tissue sectioning, immunolabeling and computer simulation to reconstruct the 3D distribution of MEPs 3, but the entire process of slicing, mounting, labeling and imaging requires a large number of manual operations, which are both time-consuming and labor- intensive, followed by an extremely complicated image registration and data loss 15.…”

Section: Introductionmentioning

confidence: 99%

Spatial Distribution of Motor Endplates and its Adaptive Change in Skeletal Muscle

Yin

Chen

et al. 2019

Theranostics

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Motor endplates (MEPs) are the important interfaces between peripheral nerves and muscle fibers. Investigation of the spatial distribution of MEPs could help us better understand neuromuscular functional activities and improve the diagnosis and therapy of related diseases. Methods: Fluorescent α-bungarotoxin was injected to label the motor endplates in whole-mount skeletal muscles, and tissue optical clearing combined with light-sheet microscopy was used to investigate the spatial distribution of MEPs and in-muscle nerve branches in different skeletal muscles in wild-type and transgenic fluorescent mice. Electrophysiology was used to determine the relationship between the spatial distribution of MEPs and muscle function. Results: The exact three-dimensional distribution of MEPs in whole skeletal muscles was first obtained. We found that the MEPs in the muscle were distributed in an organized pattern of lamella clusters, with no MEPs outside the lamella zone. Each MEP lamella was innervated by one independent in-muscle nerve branch and mediated an independent muscle subgroup contraction. Additionally, the MEPs changed along the lamella clusters after denervation and regained the initial pattern after reinnervation. The integrity and spatial distribution of MEPs could reflect the functional state of muscles. The signal absence of a certain MEP lamella could suggest a problem in certain part of the muscle. Conclusions: The MEP lamella clusters might be the basis of neuromuscular function, and the spatial distribution of MEPs could serve as a testbed for evaluating the functional status of muscle and the therapeutic targeting map related to MEPs.

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Section: Introductionmentioning

confidence: 99%

Spatial Distribution of Motor Endplates and its Adaptive Change in Skeletal Muscle

Yin

Chen

et al. 2019

Theranostics

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Early Changes in Porcine Larynges Following Injection of Motor‐Endplate Expressing Muscle Cells for the Treatment of Unilateral Vocal Fold Paralysis

et al. 2023

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ObjectivesNo curative injectable therapy exists for unilateral vocal fold paralysis. Herein, we explore the early implications of muscle‐derived motor‐endplate expressing cells (MEEs) for injectable vocal fold medialization after recurrent laryngeal nerve (RLN) injury.MethodsYucatan minipigs underwent right RLN transection (without repair) and muscle biopsies. Autologous muscle progenitor cells were isolated, cultured, differentiated, and induced to form MEEs. Three weeks after the injury, MEEs or saline were injected into the paralyzed right vocal fold. Outcomes including evoked laryngeal electromyography (LEMG), laryngeal adductor pressure, and acoustic vocalization data were analyzed up to 7 weeks post‐injury. Harvested porcine larynges were examined for volume, gene expression, and histology.ResultsMEE injections were tolerated well, with all pigs demonstrating continued weight gain. Blinded analysis of videolaryngoscopy post‐injection revealed infraglottic fullness, and no inflammatory changes. Four weeks after injection, LEMG revealed on average higher right distal RLN activity retention in MEE pigs. MEE‐injected pigs on average had vocalization durations, frequencies, and intensities higher than saline pigs. Post‐mortem, the MEE‐injected larynges revealed statistically greater volume on quantitative 3D ultrasound, and statistically increased expression of neurotrophic factors (BDNF, NGF, NTF3, NTF4, NTN1) on quantitative PCR.ConclusionsMinimally invasive MEE injection appears to establish an early molecular and microenvironmental framework to encourage innate RLN regeneration. Longer follow‐up is needed to determine if early findings will translate into functional contraction.Level of EvidenceNA Laryngoscope, 2023

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The Yucatan Minipig: A Reliable Model for Studying Unilateral Vocal Fold Paralysis

Kaefer,

Calcagno,

Feucht

et al. 2024

Journal of Voice

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Immunohistochemical Detection of Motor Endplates in the Long-Term Denervated Muscle

Abstract: Persistence of MEPs in the long-term denervated SM suggests that some surgeries targeting the MEPs such as NMEG-NMZ technique should be effective for delayed reinnervation. However, more work is needed to develop strategies for preservation of muscle mass and MEPs after denervation.

Cited by 6 publications

References 71 publications

Spatial Distribution of Motor Endplates and its Adaptive Change in Skeletal Muscle

Spatial Distribution of Motor Endplates and its Adaptive Change in Skeletal Muscle

Early Changes in Porcine Larynges Following Injection of Motor‐Endplate Expressing Muscle Cells for the Treatment of Unilateral Vocal Fold Paralysis

The Yucatan Minipig: A Reliable Model for Studying Unilateral Vocal Fold Paralysis

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