Abstract
The neural retina is a critical component of the visual system, which provides the majority of sensory input in humans. Various retinal degenerative diseases can result in the permanent loss of retinal neurons, especially the light-sensing photoreceptors and the centrally projecting retinal ganglion cells (RGCs). The replenishment of lost RGCs and the repair of optic nerve damage are particularly challenging, as both RGC specification and their subsequent axonal growth and projection involve complex and precise regulation. To explore the developmental potential of pluripotent stem cell-derived neural progenitors, we have established mouse iPS cells that allow cell lineage tracing of progenitors that have expressed Atoh7/Math5, a bHLH transcription factor required for RGC production. These Atoh7 lineage reporter iPS cells encode Cre to replace one copy of the endogenous Atoh7 gene and a Cre-dependent YFP reporter in the ROSA locus. In addition, they express pluripotent markers and are capable of generating teratomas in vivo. Under anterior neural induction and neurogenic conditions in vitro, the Atoh7-Cre/ROSA-YFP iPS cells differentiate into neurons that co-express various RGC markers and YFP, indicating that these neurons are derived from Atoh7-expressing progenitors. Consistent with previous in vivo cell lineage studies, the Atoh7-Cre/ROSA-YFP iPS cells also give rise to a subset of Crx-positive photoreceptor precursors. Furthermore, inhibition of Notch signaling in the iPSC cultures results in a significant increase of YFP-positive RGCs and photoreceptor precursors. Together, these results show that Atoh7-Cre/ROSA-YFP iPS cells can be used to monitor the development and survival of RGCs and photoreceptors from pluripotent stem cells.
Highlights
The neural retina is a component of the central nervous system and plays an essential role in the acquisition and processing of visual information
Generation of Atoh7-Cre/ROSA-YFP induced pluripotent stem (iPS) cells Since Atoh7 is transiently expressed during retinogenesis and is no longer expressed by differentiated retinal ganglion cells (RGCs), we sought to establish iPS cells that will allow us to monitor Atoh7 expression as well as trace the Atoh7 lineage, including mature RGCs
Repairing vision impairment associated with axonal damage and/or the loss of RGCs is challenging, as RGCs emerge during early retinogenesis and connect to retinal interneurons and extend centrally projecting axons along complex trajectory routes to establish functional visual circuitry
Summary
The neural retina is a component of the central nervous system and plays an essential role in the acquisition and processing of visual information. The mature retina consists of distinct neuronal cell types derived from a common pool of neural progenitor cells during development [1,2,3]. De novo production of distinct retinal neurons, especially photoreceptor cells and RGCs, for the purposes of repairing damaged retinas as well as enabling disease mechanism studies remains a high priority. The production of RGCs from the retinal primordium is stringently controlled by cell-intrinsic transcription factors and influenced by cell-extrinsic signals. RGC production is negatively regulated by Notch signaling and a number of RGC-derived secreted factors, including GDF11, Sonic Hedgehog (Shh), and VEGF [24,25,26,27,28,29,30]. Genetic disruption of Notch, GDF11, or Shh signaling increases Atoh expression and enhances RGC and cone photoreceptor genesis [26,28,29,31,32,33]
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