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In Vivo Direct Reprogramming of Resident Glial Cells into Interneurons by Intracerebral Injection of Viral Vectors

Pereira, Maria LU ; Birtele, Marcella LU and Rylander Ottosson, Daniella LU (2019) In Journal of visualized experiments : JoVE
Abstract

Converting resident glia in the brain into functional and subtype-specific neurons in vivo provides a step forward towards the development of alternative cell replacement therapies while also creating tools to study cell fate in situ. To date, it has been possible to obtain neurons via in vivo reprogramming, but the precise phenotype of these neurons or how they mature has not been analyzed in detail. In this protocol, we describe a more efficient conversion and cell-specific identification of the in vivo reprogrammed neurons, using an AAV-based viral vector system. We also provide a protocol for functional assessment of the reprogrammed cells' neuronal maturation. By injecting flip-excision (FLEX) vectors, containing the reprogramming... (More)

Converting resident glia in the brain into functional and subtype-specific neurons in vivo provides a step forward towards the development of alternative cell replacement therapies while also creating tools to study cell fate in situ. To date, it has been possible to obtain neurons via in vivo reprogramming, but the precise phenotype of these neurons or how they mature has not been analyzed in detail. In this protocol, we describe a more efficient conversion and cell-specific identification of the in vivo reprogrammed neurons, using an AAV-based viral vector system. We also provide a protocol for functional assessment of the reprogrammed cells' neuronal maturation. By injecting flip-excision (FLEX) vectors, containing the reprogramming and synapsin-driven reporter genes to specific cell types in the brain that serve as the target for cell reprogramming. This technique allows for the easy identification of newly reprogrammed neurons. Results show that the obtained reprogrammed neurons functionally mature over time, receive synaptic contacts and show electrophysiological properties of different types of interneurons. Using the transcription factors Ascl1, Lmx1a and Nurr1, the majority of the reprogrammed cells have properties of fast-spiking, parvalbumin-containing interneurons.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of visualized experiments : JoVE
issue
148
article number
e59465
publisher
JoVE
external identifiers
  • scopus:85068974023
  • pmid:31259901
ISSN
1940-087X
DOI
10.3791/59465
language
English
LU publication?
yes
id
e11a86f8-7b2a-449c-9ec7-aee6023c589d
alternative location
https://www.jove.com/video/59465/in-vivo-direct-reprogramming-resident-glial-cells-into-interneurons
date added to LUP
2019-07-09 10:51:36
date last changed
2020-01-13 02:12:51
@article{e11a86f8-7b2a-449c-9ec7-aee6023c589d,
  abstract     = {<p>Converting resident glia in the brain into functional and subtype-specific neurons in vivo provides a step forward towards the development of alternative cell replacement therapies while also creating tools to study cell fate in situ. To date, it has been possible to obtain neurons via in vivo reprogramming, but the precise phenotype of these neurons or how they mature has not been analyzed in detail. In this protocol, we describe a more efficient conversion and cell-specific identification of the in vivo reprogrammed neurons, using an AAV-based viral vector system. We also provide a protocol for functional assessment of the reprogrammed cells' neuronal maturation. By injecting flip-excision (FLEX) vectors, containing the reprogramming and synapsin-driven reporter genes to specific cell types in the brain that serve as the target for cell reprogramming. This technique allows for the easy identification of newly reprogrammed neurons. Results show that the obtained reprogrammed neurons functionally mature over time, receive synaptic contacts and show electrophysiological properties of different types of interneurons. Using the transcription factors Ascl1, Lmx1a and Nurr1, the majority of the reprogrammed cells have properties of fast-spiking, parvalbumin-containing interneurons.</p>},
  author       = {Pereira, Maria and Birtele, Marcella and Rylander Ottosson, Daniella},
  issn         = {1940-087X},
  language     = {eng},
  month        = {06},
  number       = {148},
  publisher    = {JoVE},
  series       = {Journal of visualized experiments : JoVE},
  title        = {In Vivo Direct Reprogramming of Resident Glial Cells into Interneurons by Intracerebral Injection of Viral Vectors},
  url          = {http://dx.doi.org/10.3791/59465},
  doi          = {10.3791/59465},
  year         = {2019},
}