Like other areas of the nervous system, the retina is subject to many acquired and inherited neuronal degenerative diseases. Since the retina provides the input for all visual sensory information to the brain, the loss of cells results in viual impairment and potentially complete blindness. Many retinal degenerative diseases affect only a subset of the retinal cells, although, frequently in more advanced disease, loss and reorganization of the entire retina can occur. In mammals, there is very limited regeneration of the degenerated cells;however, in fish, new neurons of all types regenerate from M?ller glia following retinal damage and they are functionally integrated into the existing circuitry. Nevertheless, M?ller glia, the cellular source for regeneration, is present in all vertebrate retins. In the proposal we submitted three years ago, we hypothesized that regeneration from mammalian M?ller glia was limited because they fail to express a key proneural transcription factor, Ascl1, after injury. We proposed to test this hypothesis by virally-mediated expression of Ascl1 in mouse M?ller glia. In the two years of funding, we have tested the hypothesis, and found that viral expression of Ascl1 is sufficient to activate a neurogenic program in mouse M?ller glia, both in dissociated cultures and in the intact retina. The reprogrammed M?ller glia generates cells that resemble neurons in morphology, gene expression and their responses to neurotransmitters. In the next funding period, we propose to further optimize this reprogramming process, using other transcription factors and epigenetic modifiers, and then to test whether Ascl1-reprogrammed Muller glia can provide a source for regeneration in vivo in a newly developed line of transgenic mice.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
2R01EY021482-04
Application #
8690333
Study Section
Neurogenesis and Cell Fate Study Section (NCF)
Program Officer
Greenwell, Thomas
Project Start
2011-04-01
Project End
2018-03-31
Budget Start
2014-04-01
Budget End
2015-03-31
Support Year
4
Fiscal Year
2014
Total Cost
$386,250
Indirect Cost
$136,250
Name
University of Washington
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
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Wohl, Stefanie G; Reh, Thomas A (2016) The microRNA expression profile of mouse Müller glia in vivo and in vitro. Sci Rep 6:35423
Brzezinski, Joseph A; Reh, Thomas A (2015) Photoreceptor cell fate specification in vertebrates. Development 142:3263-73
Ueki, Yumi; Wilken, Matthew S; Cox, Kristen E et al. (2015) Transgenic expression of the proneural transcription factor Ascl1 in Müller glia stimulates retinal regeneration in young mice. Proc Natl Acad Sci U S A 112:13717-22
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Ueki, Yumi; Wilken, Matthew S; Cox, Kristen E et al. (2015) A transient wave of BMP signaling in the retina is necessary for Müller glial differentiation. Development 142:533-43
Vierstra, Jeff; Rynes, Eric; Sandstrom, Richard et al. (2014) Mouse regulatory DNA landscapes reveal global principles of cis-regulatory evolution. Science 346:1007-12
Cheng, Yong; Ma, Zhihai; Kim, Bong-Hyun et al. (2014) Principles of regulatory information conservation between mouse and human. Nature 515:371-5
Stergachis, Andrew B; Neph, Shane; Sandstrom, Richard et al. (2014) Conservation of trans-acting circuitry during mammalian regulatory evolution. Nature 515:365-70

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