Cargando…

Oligodendrocyte Precursor Cell Transplantation into Organotypic Cerebellar Shiverer Slices: A Model to Study Myelination and Myelin Maintenance

Current in vitro models to investigate the consequence of oligodendrocyte-specific loss-of-function mutations on myelination are primarily limited to co-culture experiments, which do not accurately recapitulate the complex in vivo environment. Here, we describe the development of an in vitro model o...

Descripción completa

Detalles Bibliográficos
Autores principales: Bin, Jenea M., Leong, Soo Yuen, Bull, Sarah-Jane, Antel, Jack P., Kennedy, Timothy E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3401123/
https://www.ncbi.nlm.nih.gov/pubmed/22911763
http://dx.doi.org/10.1371/journal.pone.0041237
_version_ 1782238569013706752
author Bin, Jenea M.
Leong, Soo Yuen
Bull, Sarah-Jane
Antel, Jack P.
Kennedy, Timothy E.
author_facet Bin, Jenea M.
Leong, Soo Yuen
Bull, Sarah-Jane
Antel, Jack P.
Kennedy, Timothy E.
author_sort Bin, Jenea M.
collection PubMed
description Current in vitro models to investigate the consequence of oligodendrocyte-specific loss-of-function mutations on myelination are primarily limited to co-culture experiments, which do not accurately recapitulate the complex in vivo environment. Here, we describe the development of an in vitro model of myelination and myelin maintenance in which oligodendrocyte precursor cells are transplanted into organotypic cerebellar slice cultures derived from dysmyelinated shiverer mice. Compared to neuron-oligodendrocyte co-cultures, organotypic slices more closely mimic the environment in vivo, while utilizing a genetic background that allows for straight-forward identification of myelin generated by transplanted cells. We show at the ultrastructural level that the myelin generated by wild-type transplanted oligodendrocytes is compact and terminates in cytoplasmic loops that form paranodal junctions with the axon. This myelination results in the appropriate sequestering of axonal proteins into specialized domains surrounding the nodes of Ranvier. We also demonstrate the applicability of this approach for xenograft transplantation of oligodendrocyte precursor cells derived from rat or human sources. This method provides a time-efficient and cost-effective adjunct to conditional knockout mouse lines or in vivo transplantation models to study oligodendrocyte-specific loss-of-function mutations. Furthermore, the approach can be readily used to assess the effect of pharmacological manipulations on myelin, providing a tool to better understand myelination and develop effective therapeutic strategies to treat myelin-related diseases.
format Online
Article
Text
id pubmed-3401123
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-34011232012-07-30 Oligodendrocyte Precursor Cell Transplantation into Organotypic Cerebellar Shiverer Slices: A Model to Study Myelination and Myelin Maintenance Bin, Jenea M. Leong, Soo Yuen Bull, Sarah-Jane Antel, Jack P. Kennedy, Timothy E. PLoS One Research Article Current in vitro models to investigate the consequence of oligodendrocyte-specific loss-of-function mutations on myelination are primarily limited to co-culture experiments, which do not accurately recapitulate the complex in vivo environment. Here, we describe the development of an in vitro model of myelination and myelin maintenance in which oligodendrocyte precursor cells are transplanted into organotypic cerebellar slice cultures derived from dysmyelinated shiverer mice. Compared to neuron-oligodendrocyte co-cultures, organotypic slices more closely mimic the environment in vivo, while utilizing a genetic background that allows for straight-forward identification of myelin generated by transplanted cells. We show at the ultrastructural level that the myelin generated by wild-type transplanted oligodendrocytes is compact and terminates in cytoplasmic loops that form paranodal junctions with the axon. This myelination results in the appropriate sequestering of axonal proteins into specialized domains surrounding the nodes of Ranvier. We also demonstrate the applicability of this approach for xenograft transplantation of oligodendrocyte precursor cells derived from rat or human sources. This method provides a time-efficient and cost-effective adjunct to conditional knockout mouse lines or in vivo transplantation models to study oligodendrocyte-specific loss-of-function mutations. Furthermore, the approach can be readily used to assess the effect of pharmacological manipulations on myelin, providing a tool to better understand myelination and develop effective therapeutic strategies to treat myelin-related diseases. Public Library of Science 2012-07-20 /pmc/articles/PMC3401123/ /pubmed/22911763 http://dx.doi.org/10.1371/journal.pone.0041237 Text en Bin et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Bin, Jenea M.
Leong, Soo Yuen
Bull, Sarah-Jane
Antel, Jack P.
Kennedy, Timothy E.
Oligodendrocyte Precursor Cell Transplantation into Organotypic Cerebellar Shiverer Slices: A Model to Study Myelination and Myelin Maintenance
title Oligodendrocyte Precursor Cell Transplantation into Organotypic Cerebellar Shiverer Slices: A Model to Study Myelination and Myelin Maintenance
title_full Oligodendrocyte Precursor Cell Transplantation into Organotypic Cerebellar Shiverer Slices: A Model to Study Myelination and Myelin Maintenance
title_fullStr Oligodendrocyte Precursor Cell Transplantation into Organotypic Cerebellar Shiverer Slices: A Model to Study Myelination and Myelin Maintenance
title_full_unstemmed Oligodendrocyte Precursor Cell Transplantation into Organotypic Cerebellar Shiverer Slices: A Model to Study Myelination and Myelin Maintenance
title_short Oligodendrocyte Precursor Cell Transplantation into Organotypic Cerebellar Shiverer Slices: A Model to Study Myelination and Myelin Maintenance
title_sort oligodendrocyte precursor cell transplantation into organotypic cerebellar shiverer slices: a model to study myelination and myelin maintenance
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3401123/
https://www.ncbi.nlm.nih.gov/pubmed/22911763
http://dx.doi.org/10.1371/journal.pone.0041237
work_keys_str_mv AT binjeneam oligodendrocyteprecursorcelltransplantationintoorganotypiccerebellarshivererslicesamodeltostudymyelinationandmyelinmaintenance
AT leongsooyuen oligodendrocyteprecursorcelltransplantationintoorganotypiccerebellarshivererslicesamodeltostudymyelinationandmyelinmaintenance
AT bullsarahjane oligodendrocyteprecursorcelltransplantationintoorganotypiccerebellarshivererslicesamodeltostudymyelinationandmyelinmaintenance
AT anteljackp oligodendrocyteprecursorcelltransplantationintoorganotypiccerebellarshivererslicesamodeltostudymyelinationandmyelinmaintenance
AT kennedytimothye oligodendrocyteprecursorcelltransplantationintoorganotypiccerebellarshivererslicesamodeltostudymyelinationandmyelinmaintenance