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Volume 13
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10.3390/cells13171408
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Article

Comparing Viral Vectors and Fate Mapping Approaches for Astrocyte-to-Neuron Reprogramming in the Injured Mouse Cerebral Cortex

by
Matteo Puglisi
1,2,3,
Chu Lan Lao
1,2,4,
Gulzar Wani
1,2,
Giacomo Masserdotti
1,2,
Riccardo Bocchi
1,2,† and
Magdalena Götz
1,2,4,*
1
Division of Physiological Genomics, Biomedical Center, Ludwig-Maximilians-Universität München, 82152 Planegg-Martinsried, Germany
2
Institute for Stem Cell Research, Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH), 85764 Nuremberg, Germany
3
Graduate School of Systemic Neuroscience, Biomedical Center, Ludwig-Maximilians-Universität München, 82152 Planegg-Martinsried, Germany
4
Munich Cluster for Systems Neurology (SyNergy), Biomedical Center, Ludwig-Maximilians-Universität München, 82152 Planegg-Martinsried, Germany
*
Author to whom correspondence should be addressed.
†
Current address: Department of Basic Neuroscience, University of Geneva, CH-1211 Geneva, Switzerland.
Cells 2024, 13(17), 1408; https://doi.org/10.3390/cells13171408
Submission received: 31 July 2024 / Revised: 16 August 2024 / Accepted: 20 August 2024 / Published: 23 August 2024
(This article belongs to the Special Issue Astrocyte Reprogramming and Brain Homeostasis)
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Abstract

Direct neuronal reprogramming is a promising approach to replace neurons lost due to disease via the conversion of endogenous glia reacting to brain injury into neurons. However, it is essential to demonstrate that the newly generated neurons originate from glial cells and/or show that they are not pre-existing endogenous neurons. Here, we use controls for both requirements while comparing two viral vector systems (Mo-MLVs and AAVs) for the expression of the same neurogenic factor, the phosphorylation-resistant form of Neurogenin2. Our results show that Mo-MLVs targeting proliferating glial cells after traumatic brain injury reliably convert astrocytes into neurons, as assessed by genetic fate mapping of astrocytes. Conversely, expressing the same neurogenic factor in a flexed AAV system results in artefactual labelling of endogenous neurons fatemapped by birthdating in development that are negative for the genetic fate mapping marker induced in astrocytes. These results are further corroborated by chronic live in vivo imaging. Taken together, the phosphorylation-resistant form of Neurogenin2 is more efficient in reprogramming reactive glia into neurons than its wildtype counterpart in vivo using retroviral vectors (Mo-MLVs) targeting proliferating glia. Conversely, AAV-mediated expression generates artefacts and is not sufficient to achieve fate conversion.
Keywords: astrocytes; neurons; direct reprogramming; fate mapping; birthdating; viral vectors; retrovirus; AAV; Neurogenin2 astrocytes; neurons; direct reprogramming; fate mapping; birthdating; viral vectors; retrovirus; AAV; Neurogenin2

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MDPI and ACS Style

Puglisi, M.; Lao, C.L.; Wani, G.; Masserdotti, G.; Bocchi, R.; Götz, M. Comparing Viral Vectors and Fate Mapping Approaches for Astrocyte-to-Neuron Reprogramming in the Injured Mouse Cerebral Cortex. Cells 2024, 13, 1408. https://doi.org/10.3390/cells13171408

AMA Style

Puglisi M, Lao CL, Wani G, Masserdotti G, Bocchi R, Götz M. Comparing Viral Vectors and Fate Mapping Approaches for Astrocyte-to-Neuron Reprogramming in the Injured Mouse Cerebral Cortex. Cells. 2024; 13(17):1408. https://doi.org/10.3390/cells13171408

Chicago/Turabian Style

Puglisi, Matteo, Chu Lan Lao, Gulzar Wani, Giacomo Masserdotti, Riccardo Bocchi, and Magdalena Götz. 2024. "Comparing Viral Vectors and Fate Mapping Approaches for Astrocyte-to-Neuron Reprogramming in the Injured Mouse Cerebral Cortex" Cells 13, no. 17: 1408. https://doi.org/10.3390/cells13171408

APA Style

Puglisi, M., Lao, C. L., Wani, G., Masserdotti, G., Bocchi, R., & Götz, M. (2024). Comparing Viral Vectors and Fate Mapping Approaches for Astrocyte-to-Neuron Reprogramming in the Injured Mouse Cerebral Cortex. Cells, 13(17), 1408. https://doi.org/10.3390/cells13171408

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