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A Novel Bioengineered Functional Motor Unit Platform to Study Neuromuscular Interaction

Background: In many neurodegenerative and muscular disorders, and loss of innervation in sarcopenia, improper reinnervation of muscle and dysfunction of the motor unit (MU) are key pathogenic features. In vivo studies of MUs are constrained due to difficulties isolating and extracting functional MUs...

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Autores principales: Saini, Jasdeep, Faroni, Alessandro, Reid, Adam J., Mamchaoui, Kamel, Mouly, Vincent, Butler-Browne, Gillian, Lightfoot, Adam P., McPhee, Jamie S., Degens, Hans, Al-Shanti, Nasser
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7599749/
https://www.ncbi.nlm.nih.gov/pubmed/33050427
http://dx.doi.org/10.3390/jcm9103238
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author Saini, Jasdeep
Faroni, Alessandro
Reid, Adam J.
Mamchaoui, Kamel
Mouly, Vincent
Butler-Browne, Gillian
Lightfoot, Adam P.
McPhee, Jamie S.
Degens, Hans
Al-Shanti, Nasser
author_facet Saini, Jasdeep
Faroni, Alessandro
Reid, Adam J.
Mamchaoui, Kamel
Mouly, Vincent
Butler-Browne, Gillian
Lightfoot, Adam P.
McPhee, Jamie S.
Degens, Hans
Al-Shanti, Nasser
author_sort Saini, Jasdeep
collection PubMed
description Background: In many neurodegenerative and muscular disorders, and loss of innervation in sarcopenia, improper reinnervation of muscle and dysfunction of the motor unit (MU) are key pathogenic features. In vivo studies of MUs are constrained due to difficulties isolating and extracting functional MUs, so there is a need for a simplified and reproducible system of engineered in vitro MUs. Objective: to develop and characterise a functional MU model in vitro, permitting the analysis of MU development and function. Methods: an immortalised human myoblast cell line was co-cultured with rat embryo spinal cord explants in a serum-free/growth fact media. MUs developed and the morphology of their components (neuromuscular junction (NMJ), myotubes and motor neurons) were characterised using immunocytochemistry, phase contrast and confocal microscopy. The function of the MU was evaluated through live observations and videography of spontaneous myotube contractions after challenge with cholinergic antagonists and glutamatergic agonists. Results: blocking acetylcholine receptors with α-bungarotoxin resulted in complete, cessation of myotube contractions, which was reversible with tubocurarine. Furthermore, myotube activity was significantly higher with the application of L-glutamic acid. All these observations indicate the formed MU are functional. Conclusion: a functional nerve-muscle co-culture model was established that has potential for drug screening and pathophysiological studies of neuromuscular interactions.
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spelling pubmed-75997492020-11-01 A Novel Bioengineered Functional Motor Unit Platform to Study Neuromuscular Interaction Saini, Jasdeep Faroni, Alessandro Reid, Adam J. Mamchaoui, Kamel Mouly, Vincent Butler-Browne, Gillian Lightfoot, Adam P. McPhee, Jamie S. Degens, Hans Al-Shanti, Nasser J Clin Med Article Background: In many neurodegenerative and muscular disorders, and loss of innervation in sarcopenia, improper reinnervation of muscle and dysfunction of the motor unit (MU) are key pathogenic features. In vivo studies of MUs are constrained due to difficulties isolating and extracting functional MUs, so there is a need for a simplified and reproducible system of engineered in vitro MUs. Objective: to develop and characterise a functional MU model in vitro, permitting the analysis of MU development and function. Methods: an immortalised human myoblast cell line was co-cultured with rat embryo spinal cord explants in a serum-free/growth fact media. MUs developed and the morphology of their components (neuromuscular junction (NMJ), myotubes and motor neurons) were characterised using immunocytochemistry, phase contrast and confocal microscopy. The function of the MU was evaluated through live observations and videography of spontaneous myotube contractions after challenge with cholinergic antagonists and glutamatergic agonists. Results: blocking acetylcholine receptors with α-bungarotoxin resulted in complete, cessation of myotube contractions, which was reversible with tubocurarine. Furthermore, myotube activity was significantly higher with the application of L-glutamic acid. All these observations indicate the formed MU are functional. Conclusion: a functional nerve-muscle co-culture model was established that has potential for drug screening and pathophysiological studies of neuromuscular interactions. MDPI 2020-10-10 /pmc/articles/PMC7599749/ /pubmed/33050427 http://dx.doi.org/10.3390/jcm9103238 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Saini, Jasdeep
Faroni, Alessandro
Reid, Adam J.
Mamchaoui, Kamel
Mouly, Vincent
Butler-Browne, Gillian
Lightfoot, Adam P.
McPhee, Jamie S.
Degens, Hans
Al-Shanti, Nasser
A Novel Bioengineered Functional Motor Unit Platform to Study Neuromuscular Interaction
title A Novel Bioengineered Functional Motor Unit Platform to Study Neuromuscular Interaction
title_full A Novel Bioengineered Functional Motor Unit Platform to Study Neuromuscular Interaction
title_fullStr A Novel Bioengineered Functional Motor Unit Platform to Study Neuromuscular Interaction
title_full_unstemmed A Novel Bioengineered Functional Motor Unit Platform to Study Neuromuscular Interaction
title_short A Novel Bioengineered Functional Motor Unit Platform to Study Neuromuscular Interaction
title_sort novel bioengineered functional motor unit platform to study neuromuscular interaction
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7599749/
https://www.ncbi.nlm.nih.gov/pubmed/33050427
http://dx.doi.org/10.3390/jcm9103238
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