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.
(Less)
- author
- Pereira, Maria
LU
; Birtele, Marcella
LU
and Rylander Ottosson, Daniella
LU
- organization
- publishing date
- 2019-06-17
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of visualized experiments : JoVE
- issue
- 148
- article number
- e59465
- publisher
- JoVE
- external identifiers
-
- pmid:31259901
- scopus:85068974023
- 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
- 2024-02-15 17:13:21
@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}},
}