Abstract
PNU-282987, a selective alpha7 nicotinic acetylcholine receptor agonist, has previously been shown to have both neurogenic and broad regenerative effects in the adult murine retina. The objective of this study was to assay the molecular mechanism by which PNU-282987 promotes the production of Muller-derived progenitor cells through signaling via the resident retinal pigment epithelium. These Muller-derived progenitor cells generate a myriad of differentiated neurons throughout the retina that have previously been characterized by morphology. Herein, we demonstrate that topical application of PNU-282987 stimulates production of functional neurons as measured by electroretinograms. Further, we examine the mechanism of how this phenomenon occurs through activation of this atypical receptor using a transcriptomic approach isolated retinal pigment epithelium activated by PNU-282987 and in whole retina. We provide evidence that PNU-282987 causes a bi-modal signaling event in which early activation primes the retina with an inflammatory response and developmental signaling cues, followed by an inhibition of gliotic mechanisms and a decrease in the immune response, ending with upregulation of genes associated with specific retinal neuron generation. Taken together, these data provide evidence that PNU-282987 activates the retinal pigment epithelium to signal to Muller glia to produce Muller-derived progenitor cells, which can differentiate into new, functional neurons in adult mice. These data not only increase our understanding of how adult mammalian retinal regeneration can occur, but also provide therapeutic promise for treating functional vision loss.
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Data availability
RNAseq dataset generated has been deposited to NCBI GEO GSE156758.
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Acknowledgements
A special thanks to J. Duncan for his expert consultation and access to qRT-PCR equipment as well as to M. Stanchfield for assistance with cell culture maintenance and to E. Levine for use of the transgenic mice he developed.
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This study was funded by NIH #EY027970 to C. Linn.
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SEW designed and performed all experiments, generated data and figures, and wrote the manuscript. JBS performed the ERG experiments. MKW assisted in the design of the genome editing scheme and with bioinformatics. DL performed the intravitreal injections and aided in the writing of the manuscript. DCO contributed to the study design and writing the manuscript. CTT generated whole retina RNAseq dataset. CL acted as project manager, edited the final manuscript and obtained the funding for this project.
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Webster, S.E., Spitsbergen, J.B., Linn, D.M. et al. Transcriptome Changes in Retinal Pigment Epithelium Post-PNU-282987 Treatment Associated with Adult Retinal Neurogenesis in Mice. J Mol Neurosci 72, 1990–2010 (2022). https://doi.org/10.1007/s12031-022-02049-z
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DOI: https://doi.org/10.1007/s12031-022-02049-z