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Insm1a-mediated gene repression is essential for the formation and differentiation of Müller glia-derived progenitors in the injured retina

Nature Cell Biology volume 14, pages 1013–1023 (2012)Cite this article

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Abstract

In zebrafish, retinal injury stimulates Müller glia (MG) reprograming, allowing them to generate multipotent progenitors that replace damaged cells and restore vision. Recent studies suggest that transcriptional repression may underlie these events. To identify transcriptional repressors, we compared the transcriptomes of MG and MG-derived progenitors and identified insm1a, a repressor exhibiting a biphasic pattern of expression that is essential for retina regeneration. Insm1a was found to suppress ascl1a and its own expression, and link injury-dependent ascl1a induction with the suppression of the Wnt inhibitor dickkopf (dkk), which is necessary for MG dedifferentiation. We also found that Insm1a was responsible for sculpting the zone of injury-responsive MG by suppressing h b-e g fa expression. Finally, we provide evidence that Insm1a stimulates progenitor cell-cycle exit by suppressing a genetic program driving progenitor proliferation. Our studies identify Insm1a as a key regulator of retina regeneration and provide a mechanistic understanding of how it contributes to multiple phases of this process.

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Figure 1: Insm1a is necessary for MG dedifferentiation in the injured retina.
Figure 2: An Ascl1a–Insm1a regulatory loop.
Figure 3: Insm1a regulates dkk promoter activity.
Figure 4: Insm1a regulates the zone of injury-responsive MG.
Figure 5: Insm1a restricts the zone of dedifferentiating MG by suppressing h b-e g fa gene expression.
Figure 6: At 4–6 dpi Insm1a inhibits a genetic program driving cell proliferation and stimulates a genetic program driving cell-cycle exit.
Figure 7: Insm1a stimulates p57kip2 expression by suppressing bcl11 gene expression.
Figure 8: Insm1a knockdown transiently suppresses progenitor differentiation.

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Acknowledgements

This research was supported by NEI grant RO1 EY 018132 from the NIH. We thank D. Hyde (University of Notre Dame, USA) for gfap:gfp transgenic fish; R. Moon (University of Washington, USA) for hsp70:dkk1b-gfp transgenic fish; the University of Michigan Flow Cytometry Core for cell sorting; A. Dombrowski (Wayne State, USA) for microarray screen; R. Thompson (University of Michigan, USA) for assistance with microarray data organization; M. Uhler and D. Turner (University of Michigan, USA) for providing the pEL and pCS2 vectors, respectively; J. Beals (University of Michigan, USA) for help with confocal microscopy; the Goldman laboratory for helpful comments and suggestions during the course of this research; and R. Karr for fish care.

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  1. Molecular and Behavioral Neuroscience Institute and Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA

    Rajesh Ramachandran, Xiao-Feng Zhao & Daniel Goldman

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  1. Rajesh Ramachandran
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  2. Xiao-Feng Zhao
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Contributions

D.G. and R.R. conceived the study and designed experiments. R.R. and X-F.Z. performed the experiments. R.R., X-F.Z. and D.G. analysed the data and wrote the manuscript.

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Correspondence to Daniel Goldman.

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The authors declare no competing financial interests.

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Ramachandran, R., Zhao, XF. & Goldman, D. Insm1a-mediated gene repression is essential for the formation and differentiation of Müller glia-derived progenitors in the injured retina. Nat Cell Biol 14, 1013–1023 (2012). https://doi.org/10.1038/ncb2586

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