In most vertebrates, the adult retina is unable to regenerate lost neurons, which results in loss of vision. In contrast, the zebrafish retina possesses the ability to regenerate any class of retinal neurons that are lost due to a variety of insults. For example, constant intense light causes rod and cone photoreceptor death. Photoreceptor regeneration originates from the Muller glia that reenter the cell cycle and divide to produce neuronal progenitors, which continue to proliferate and then differentiate into the regenerated neurons. The human retina also possesses Muller glia, but fails to regenerate any retinal cell class. We are interested in identifying the processes that regulate retinal regeneration of photoreceptors in the light-damaged zebrafish retina. In this proposal, we will investigate the molecular mechanisms that generate and maintain the neuronal progenitor cell population in an undifferentiated and proliferating state during regeneration, which will identify approaches to induce a full regeneration response in the damaged mammalian retina. This could provide a strategy to restore vision to individuals who suffer from genetic forms of blindness, such as retinitis pigmentosa or macular degeneration. We will explore the roles of the Pax6a, Pax6b, and Olig2 proteins during proliferation of neuronal progenitors in the light-damaged zebrafish retina. We will use a technique that we developed to electroporate antisense morpholinos into the regenerating retina. This technique gives us the powerful ability to conditionally block the translation of specific proteins during regeneration of the light-damaged retina. We will test the hypothesis that the Pax6b protein is required for the initiation of neuronal progenitor cell proliferation, but is not required for the subsequent expression of progenitor cell transcription factors, and ultimately, photoreceptor cell opsins (S. A. 1). We will then determine if Pax6a is required for the continued proliferation of the neuronal progenitors and the transcription of the olig2 gene (S. A. 2). Finally, we will determine if Olig2 is also required for the continued proliferation of the neuronal progenitor cells and, if, in the absence of Olig2, the neuronal progenitors prematurely differentiate into photoreceptors at the expense of Muller glial cells (S. A. 3).
This proposal explores key genetic checkpoints that are required to regenerate retinal neurons in the adult eye. Uncovering the molecular processes that generate neuronal progenitor cells from the Muller glia and stimulating these progenitors to proliferate until they must differentiate into the missing neurons may reveal approaches to induce neuronal regeneration from the Muller glia in the damaged human retina. Ultimately, these results may lead to therapies for different forms of human blindness.
|Gorsuch, Ryne A; Lahne, Manuela; Yarka, Clare E et al. (2017) Sox2 regulates Müller glia reprogramming and proliferation in the regenerating zebrafish retina via Lin28 and Ascl1a. Exp Eye Res 161:174-192|
|Lahne, Manuela; Gorsuch, Ryne A; Nelson, Craig M et al. (2017) Culture of Adult Transgenic Zebrafish Retinal Explants for Live-cell Imaging by Multiphoton Microscopy. J Vis Exp :|
|Campbell, Leah J; Hyde, David R (2017) Opportunities for CRISPR/Cas9 Gene Editing in Retinal Regeneration Research. Front Cell Dev Biol 5:99|
|Lahne, Manuela; Li, Jingling; Marton, Rebecca M et al. (2015) Actin-Cytoskeleton- and Rock-Mediated INM Are Required for Photoreceptor Regeneration in the Adult Zebrafish Retina. J Neurosci 35:15612-34|
|Rajaram, Kamya; Harding, Rachel L; Bailey, Travis et al. (2014) Dynamic miRNA expression patterns during retinal regeneration in zebrafish: reduced dicer or miRNA expression suppresses proliferation of Müller glia-derived neuronal progenitor cells. Dev Dyn 243:1591-605|
|Rajaram, Kamya; Harding, Rachel L; Hyde, David R et al. (2014) miR-203 regulates progenitor cell proliferation during adult zebrafish retina regeneration. Dev Biol 392:393-403|
|Gorsuch, Ryne A; Hyde, David R (2014) Regulation of Müller glial dependent neuronal regeneration in the damaged adult zebrafish retina. Exp Eye Res 123:131-40|
|Sun, Lei; Li, Ping; Carr, Aprell L et al. (2014) Transcription of the SCL/TAL1 interrupting Locus (Stil) is required for cell proliferation in adult Zebrafish Retinas. J Biol Chem 289:6934-40|
|Bailey, Travis J; Hyde, David R (2014) The phenotype of the good effort mutant zebrafish is retinal degeneration by cell death and is linked to the chromosome assembly factor 1b gene. Adv Exp Med Biol 801:535-41|
|Xu, Jia-Xi; Si, Man; Zhang, Hui-Ran et al. (2014) Phosphoinositide kinases play key roles in norepinephrine- and angiotensin II-induced increase in phosphatidylinositol 4,5-bisphosphate and modulation of cardiac function. J Biol Chem 289:6941-8|
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