The mouse levator auris longus muscle: an amenable model system to study the role of postsynaptic proteins to the maintenance and regeneration of the neuromuscular synapse

The neuromuscular junction (NMJ) is the peripheral synapse that controls the coordinated movement of many organisms. The NMJ is also an archetypical model to study synaptic morphology and function. As the NMJ is the primary target of neuromuscular diseases and traumatic injuries, the establishment o...

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Autores: Ojeda, Jorge, Bermedo-García, Francisca, Pérez, Viviana, Mella, Jessica, Hanna, Patricia, Herzberg, Daniel, Tejero, Rocío, López Manzaneda, Mario, Tabares, Lucía, Henríquez, Juan Pablo
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2020
País:España
Institución:Universidad de Sevilla (US)
Repositorio:idUS. Depósito de Investigación de la Universidad de Sevilla
OAI Identifier:oai:idus.us.es:11441/143930
Acceso en línea:https://hdl.handle.net/11441/143930
https://doi.org/10.3389/fncel.2020.00225
Access Level:acceso abierto
Palabra clave:Neuromuscular junction
Presynaptic
Postsynaptic
Regeneration
Electroporation
Skeletal muscle
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spelling The mouse levator auris longus muscle: an amenable model system to study the role of postsynaptic proteins to the maintenance and regeneration of the neuromuscular synapseOjeda, JorgeBermedo-García, FranciscaPérez, VivianaMella, JessicaHanna, PatriciaHerzberg, DanielTejero, RocíoLópez Manzaneda, MarioTabares, LucíaHenríquez, Juan PabloNeuromuscular junctionPresynapticPostsynapticRegenerationElectroporationSkeletal muscleThe neuromuscular junction (NMJ) is the peripheral synapse that controls the coordinated movement of many organisms. The NMJ is also an archetypical model to study synaptic morphology and function. As the NMJ is the primary target of neuromuscular diseases and traumatic injuries, the establishment of suitable models to study the contribution of specific postsynaptic muscle-derived proteins on NMJ maintenance and regeneration is a permanent need. Considering the unique experimental advantages of the levator auris longus (LAL) muscle, here we present a method allowing for efficient electroporation-mediated gene transfer and subsequent detailed studies of the morphology and function of the NMJ and muscle fibers. Also, we have standardized efficient facial nerve injury protocols to analyze LAL muscle NMJ degeneration and regeneration. Our results show that the expression of a control fluorescent protein does not alter either the muscle structural organization, the apposition of the pre- and post-synaptic domains, or the functional neurotransmission parameters of the LAL muscle NMJs; in turn, the overexpression of MuSK, a major regulator of postsynaptic assembly, induces the formation of ectopic acetylcholine receptor clusters. Our NMJ denervation experiments showed complete reinnervation of LAL muscle NMJs four weeks after facial nerve injury. Together, these experimental strategies in the LAL muscle constitute effective methods to combine protein expression with accurate analyses at the levels of structure, function, and regeneration of the NMJ.Fondo Nacional de Desarrollo Cientifico y Tecnologico (FONDECYT) 1170614Fondo Nacional de Desarrollo Cientifico y Tecnologico (FONDECYT) 1130321Fondo Nacional de Desarrollo Cientifico y Tecnologico (FONDECYT) 3190255Fondo Nacional de Desarrollo Cientifico y Tecnologico (FONDECYT) 3170464Ministerio de Economia, Industria y Competitividad, Gobierno de Espana/FEDER BFU201678934-PFrontiers Media S.A.Fisiología Médica y BiofísicaBIO209: Neurotransmisión y Sinaptopatologías2020info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttps://hdl.handle.net/11441/143930https://doi.org/10.3389/fncel.2020.00225reponame:idUS. Depósito de Investigación de la Universidad de Sevillainstname:Universidad de Sevilla (US)InglésFrontiers in Cellular Neuroscience, 14, 1-13.1170614; 1130321; 3190255; 3170464BFU201678934-Phttps://www.frontiersin.org/articles/10.3389/fncel.2020.00225/fullinfo:eu-repo/semantics/openAccessoai:idus.us.es:11441/1439302026-06-17T12:51:07Z
dc.title.none.fl_str_mv The mouse levator auris longus muscle: an amenable model system to study the role of postsynaptic proteins to the maintenance and regeneration of the neuromuscular synapse
title The mouse levator auris longus muscle: an amenable model system to study the role of postsynaptic proteins to the maintenance and regeneration of the neuromuscular synapse
spellingShingle The mouse levator auris longus muscle: an amenable model system to study the role of postsynaptic proteins to the maintenance and regeneration of the neuromuscular synapse
Ojeda, Jorge
Neuromuscular junction
Presynaptic
Postsynaptic
Regeneration
Electroporation
Skeletal muscle
title_short The mouse levator auris longus muscle: an amenable model system to study the role of postsynaptic proteins to the maintenance and regeneration of the neuromuscular synapse
title_full The mouse levator auris longus muscle: an amenable model system to study the role of postsynaptic proteins to the maintenance and regeneration of the neuromuscular synapse
title_fullStr The mouse levator auris longus muscle: an amenable model system to study the role of postsynaptic proteins to the maintenance and regeneration of the neuromuscular synapse
title_full_unstemmed The mouse levator auris longus muscle: an amenable model system to study the role of postsynaptic proteins to the maintenance and regeneration of the neuromuscular synapse
title_sort The mouse levator auris longus muscle: an amenable model system to study the role of postsynaptic proteins to the maintenance and regeneration of the neuromuscular synapse
dc.creator.none.fl_str_mv Ojeda, Jorge
Bermedo-García, Francisca
Pérez, Viviana
Mella, Jessica
Hanna, Patricia
Herzberg, Daniel
Tejero, Rocío
López Manzaneda, Mario
Tabares, Lucía
Henríquez, Juan Pablo
author Ojeda, Jorge
author_facet Ojeda, Jorge
Bermedo-García, Francisca
Pérez, Viviana
Mella, Jessica
Hanna, Patricia
Herzberg, Daniel
Tejero, Rocío
López Manzaneda, Mario
Tabares, Lucía
Henríquez, Juan Pablo
author_role author
author2 Bermedo-García, Francisca
Pérez, Viviana
Mella, Jessica
Hanna, Patricia
Herzberg, Daniel
Tejero, Rocío
López Manzaneda, Mario
Tabares, Lucía
Henríquez, Juan Pablo
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Fisiología Médica y Biofísica
BIO209: Neurotransmisión y Sinaptopatologías
dc.subject.none.fl_str_mv Neuromuscular junction
Presynaptic
Postsynaptic
Regeneration
Electroporation
Skeletal muscle
topic Neuromuscular junction
Presynaptic
Postsynaptic
Regeneration
Electroporation
Skeletal muscle
description The neuromuscular junction (NMJ) is the peripheral synapse that controls the coordinated movement of many organisms. The NMJ is also an archetypical model to study synaptic morphology and function. As the NMJ is the primary target of neuromuscular diseases and traumatic injuries, the establishment of suitable models to study the contribution of specific postsynaptic muscle-derived proteins on NMJ maintenance and regeneration is a permanent need. Considering the unique experimental advantages of the levator auris longus (LAL) muscle, here we present a method allowing for efficient electroporation-mediated gene transfer and subsequent detailed studies of the morphology and function of the NMJ and muscle fibers. Also, we have standardized efficient facial nerve injury protocols to analyze LAL muscle NMJ degeneration and regeneration. Our results show that the expression of a control fluorescent protein does not alter either the muscle structural organization, the apposition of the pre- and post-synaptic domains, or the functional neurotransmission parameters of the LAL muscle NMJs; in turn, the overexpression of MuSK, a major regulator of postsynaptic assembly, induces the formation of ectopic acetylcholine receptor clusters. Our NMJ denervation experiments showed complete reinnervation of LAL muscle NMJs four weeks after facial nerve injury. Together, these experimental strategies in the LAL muscle constitute effective methods to combine protein expression with accurate analyses at the levels of structure, function, and regeneration of the NMJ.
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/11441/143930
https://doi.org/10.3389/fncel.2020.00225
url https://hdl.handle.net/11441/143930
https://doi.org/10.3389/fncel.2020.00225
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Frontiers in Cellular Neuroscience, 14, 1-13.
1170614; 1130321; 3190255; 3170464
BFU201678934-P
https://www.frontiersin.org/articles/10.3389/fncel.2020.00225/full
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Frontiers Media S.A.
publisher.none.fl_str_mv Frontiers Media S.A.
dc.source.none.fl_str_mv reponame:idUS. Depósito de Investigación de la Universidad de Sevilla
instname:Universidad de Sevilla (US)
instname_str Universidad de Sevilla (US)
reponame_str idUS. Depósito de Investigación de la Universidad de Sevilla
collection idUS. Depósito de Investigación de la Universidad de Sevilla
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,300724

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