Genetic loss of function of Ptbp1 does not induce glia-to-neuron conversion in retina

Thanh Hoang; Dong Won Kim; Haley Appel; Nicole A Pannullo; Patrick Leavey; Manabu Ozawa; Sika Zheng; Minzhong Yu; Neal S Peachey; Seth Blackshaw

doi:10.1016/j.celrep.2022.110849

Genetic loss of function of Ptbp1 does not induce glia-to-neuron conversion in retina

Cell Rep. 2022 Jun 14;39(11):110849. doi: 10.1016/j.celrep.2022.110849.

Authors

Affiliations

¹ Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: vhoang3@jhmi.edu.
² Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: dkim195@jhmi.edu.
³ Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
⁴ Center for Experimental Medicine and Systems Biology, University of Tokyo, Tokyo, Japan.
⁵ Division of Biomedical Sciences, University of California, Riverside, CA, USA.
⁶ Department of Ophthalmic Research, Cole Eye Institute, Cleveland, OH, USA; Department of Ophthalmology, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, OH, USA.
⁷ Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Kavli Neuroscience Discovery Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: sblack@jhmi.edu.

Abstract

Direct reprogramming of glia into neurons is a potentially promising approach for the replacement of neurons lost to injury or neurodegenerative disorders. Knockdown of the polypyrimidine tract-binding protein Ptbp1 has been recently reported to induce efficient conversion of retinal Mϋller glia into functional neurons. Here, we use a combination of genetic lineage tracing, single-cell RNA sequencing (scRNA-seq), and electroretinogram analysis to show that selective induction of either heterozygous or homozygous loss-of-function mutants of Ptbp1 in adult retinal Mϋller glia does not lead to any detectable level of neuronal conversion. Only a few changes in gene expression are observed in Mϋller glia following Ptbp1 deletion, and glial identity is maintained. These findings highlight the importance of using genetic manipulation and lineage-tracing methods in studying cell-type conversion.

Keywords: CP: Cell biology; Mϋller glia; PTBP1; Ptbp1; RNA splicing; cell reprogramming; glia-to-neuron conversion; regeneration; retina; retinal ganglion cell; transdifferentiation.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Heterogeneous-Nuclear Ribonucleoproteins* / genetics
Heterogeneous-Nuclear Ribonucleoproteins* / metabolism
Neuroglia / metabolism
Neurons / metabolism
Polypyrimidine Tract-Binding Protein* / genetics
Polypyrimidine Tract-Binding Protein* / metabolism
Retina / metabolism

Substances

Heterogeneous-Nuclear Ribonucleoproteins
Polypyrimidine Tract-Binding Protein

Abstract

Publication types

MeSH terms

Substances

Grants and funding