This is a preview of subscription content, access via your institution
Access Nature and 54 other Nature Portfolio journals
Get Nature+, our best-value online-access subscription
$29.99 / 30 days
cancel any time
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Rent or buy this article
Prices vary by article type
Prices may be subject to local taxes which are calculated during checkout
All scRNA-seq data have been deposited at the Gene Expression Omnibus (GSE184933).
Codes for processing scRNA-seq files are available at GitHub (https://github.com/dkim195/Ptbp1_Nature_MA).
Qian, H. et al. Reversing a model of Parkinson’s disease with in situ converted nigral neurons. Nature 582, 550–556 (2020).
Su, M. et al. Expression specificity of GFAP transgenes. Neurochem. Res. 29, 2075–2093 (2004).
Fujita, T. et al. Neuronal transgene expression in dominant-negative SNARE mice. J. Neurosci. 34, 16594–16604 (2014).
Wang, L.-L. et al. Revisiting astrocyte to neuron conversion with lineage tracing in vivo. Cell 184, 5465–5481 (2021).
Le, N., Appel, H., Pannullo, N., Hoang, T. & Blackshaw, S. Ectopic insert-dependent neuronal expression of GFAP promoter-driven AAV constructs in adult mouse retina. Front. Cell Dev. Biol. 10, 914386 (2022).
Srinivasan, R. et al. New transgenic mouse lines for selectively targeting astrocytes and studying calcium signals in astrocyte processes in situ and in vivo. Neuron 92, 1181–1195 (2016).
Shibayama, M. et al. Polypyrimidine tract-binding protein is essential for early mouse development and embryonic stem cell proliferation. FEBS J. 276, 6658–6668 (2009).
Mo, A. et al. Epigenomic signatures of neuronal diversity in the mammalian brain. Neuron 86, 1369–1384 (2015).
McNeill, J., Rudyk, C., Hildebrand, M. E. & Salmaso, N. Ion channels and electrophysiological properties of astrocytes: implications for emergent stimulation technologies. Front. Cell. Neurosci. 15, 644126 (2021).
Chen, W., Zheng, Q., Huang, Q., Ma, S. & Li, M. Repressing PTBP1 fails to convert reactive astrocytes to dopaminergic neurons in a 6-hydroxydopamine mouse model of Parkinson’s disease. eLife 11, e75636 (2022).
Wang, L.-L. & Zhang, C.-L. Therapeutic potential of PTBP1 inhibition, if any, is not attributed to glia-to-neuron conversion. Annu. Rev. Neurosci. https://doi.org/10.1146/annurev-neuro-092822-083410 (2022).
Yang, G. et al. Ptbp1 knockdown failed to induce astrocytes to neurons in vivo. Gene Ther. https://doi.org/10.1038/s41434-023-00382-5 (2023).
Zhou, H. et al. Glia-to-neuron conversion by CRISPR-CasRx alleviates symptoms of neurological disease in mice. Cell 181, 590–603 (2020).
We thank F. Zhou, A. Fischer, J. Ling, C. Qian and W. Yap for their comments on the manuscript; the staff at the Single Cell & Transcriptomics Core (Johns Hopkins) for sequencing of scRNA-seq libraries. Schematics in Figs. 1 and 2, Extended Data Fig. 3 and Supplementary Figs. 1–6 were created in part using BioRender. This work was supported by the NIH National Eye Institute grants R01EY020560 and U01EY027267 to S.B.; R24EY027283 to S.B.; and the Maryland Stem Cell Research Fund (2019-MSCRFF-5124) to D.W.K. S.B. is supported by a Stein Innovation Award from Research to Prevent Blindness.
S.B. is a shareholder in CDI Labs, receives research support from Genentech and is/was a consultant to Third Rock Ventures and Tenpoint Therapeutics.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Extended data figures and tables
Extended Data Fig. 1 Astrocyte-specific deletion of Ptbp1 does not lead to replenishing lost dopaminergic neurons.
a, Schematic of the experimental timeline for 6-OHDA-induced lesion of dopaminergic neurons, followed by tamoxifen-induced specific deletion of Ptbp1 and lineage tracing of astrocytes. b, Unilateral loss of Th-positive cells in the striatum and midbrain following 6-OHDA lesion. c, Representative immunostaining images of TH and GFP in the striatum and substantia nigra in an intact and lesioned area following 6-OHDA-induced lesion across genotypes. No TH/GFP-double-positive cells were observed in any of the brain regions across genotypes. Note that TH immunostaining signals are only present in non-cell bodies in the striatum (n > = 3 mice/genotype). Scale bars = 50 μm.
Extended Data Fig. 2 RNAScope showing non-astrocytic cells in the lateral ventricular/choroid plexus regions that express GFP and Otx2 mRNA.
a, Low magnification images of GFP mRNA and DAPI in Ptbp1 KO brain sections (n > = 3 mice/genotype). b, Low magnification images of Otx2 mRNA and DAPI in Ptbp1 KO brain section. c,d, High magnification images of GFP and Otx2 mRNA across genotypes. Scale bars = 50 μm.
Extended Data Fig. 3 ScRNA-Seq analysis of doublet-cells from FACS-purified GFP+ cells in mouse brain after astrocyte-specific Ptbp1 deletion.
Violin plots showing expression of neurogenic and cluster-enriched genes in all collected cells in the cortex (a), striatum (b) and substantia nigra (c) across genotypes. Note that Neurog2 and Neurod1 were not detected in the striatum and substantia nigra scRNA-Seq dataset. The drawings of coronal sections were created using BioRender (https://www.biorender.com/).
The legends for Supplementary Figs. 1–6 and Supplementary Tables 1–17, the Supplementary Methods and references.
Ptbp1 immunohistochemistry analysis in the adult mouse brain after astrocyte-specific deletion of Ptbp1. The drawings of coronal sections were created using BioRender (https://www.biorender.com/).
Immunohistochemistry analysis of the adult mouse brain using neuronal markers after astrocyte-specific deletion of Ptbp1. The drawings of coronal sections were created using BioRender (https://www.biorender.com/).
Immunohistochemistry analysis of the adult mouse brain using dopamine markers after astrocyte-specific deletion of Ptbp1. The drawings of coronal sections were created using BioRender (https://www.biorender.com/).
scRNA-seq analysis of subsets of astrocytes in the mouse brain after astrocyte-specific Ptbp1 deletion. The drawings of coronal sections were created using BioRender (https://www.biorender.com/).
scRNA-seq analysis of the mouse brain after astrocyte-specific Ptbp1 deletion. The drawings of coronal sections were created using BioRender (https://www.biorender.com/).
scRNA-seq analysis of FACS-purified GFP+ cells in the mouse brain after astrocyte-specific Ptbp1 deletion. The drawings of coronal sections were created using BioRender (https://www.biorender.com/).
About this article
Cite this article
Hoang, T., Kim, D.W., Appel, H. et al. Ptbp1 deletion does not induce astrocyte-to-neuron conversion. Nature 618, E1–E7 (2023). https://doi.org/10.1038/s41586-023-06066-9