Abstract
MicroRNA-124a (miR-124a) is the most abundant microRNA expressed in the vertebrate CNS. Despite past investigations into the role of miR-124a, inconsistent results have left the in vivo function of miR-124a unclear. We examined the in vivo function of miR-124a by targeted disruption of Rncr3 (retinal non-coding RNA 3), the dominant source of miR-124a. Rncr3−/− mice exhibited abnormalities in the CNS, including small brain size, axonal mis-sprouting of dentate gyrus granule cells and retinal cone cell death. We found that Lhx2 is an in vivo target mRNA of miR-124a. We also observed that LHX2 downregulation by miR-124a is required for the prevention of apoptosis in the developing retina and proper axonal development of hippocampal neurons. These results suggest that miR-124a is essential for the maturation and survival of dentate gyrus neurons and retinal cones, as it represses Lhx2 translation.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Lagos-Quintana, M. et al. Identification of tissue-specific microRNAs from mouse. Curr. Biol. 12, 735–739 (2002).
Tabarés-Seisdedos, R. & Rubenstein, J.L. Chromosome 8p as a potential hub for developmental neuropsychiatric disorders: implications for schizophrenia, autism and cancer. Mol. Psychiatry 14, 563–589 (2009).
Lim, L.P. et al. Microarray analysis shows that some microRNAs downregulate large numbers of target mRNAs. Nature 433, 769–773 (2005).
Visvanathan, J., Lee, S., Lee, B., Lee, J.W. & Lee, S.K. The microRNA miR-124 antagonizes the anti-neural REST/SCP1 pathway during embryonic CNS development. Genes Dev. 21, 744–749 (2007).
Makeyev, E.V., Zhang, J., Carrasco, M.A. & Maniatis, T. The microRNA miR-124 promotes neuronal differentiation by triggering brain-specific alternative pre-mRNA splicing. Mol. Cell 27, 435–448 (2007).
Cheng, L.C., Pastrana, E., Tavazoie, M. & Doetsch, F. miR-124 regulates adult neurogenesis in the subventricular zone stem cell niche. Nat. Neurosci. 12, 399–408 (2009).
Cao, X., Pfaff, S.L. & Gage, F.H. A functional study of miR-124 in the developing neural tube. Genes Dev. 21, 531–536 (2007).
De Pietri Tonelli, D. et al. miRNAs are essential for survival and differentiation of newborn neurons but not for expansion of neural progenitors during early neurogenesis in the mouse embryonic neocortex. Development 135, 3911–3921 (2008).
Georgi, S.A. & Reh, T.A. Dicer is required for the transition from early to late progenitor state in the developing mouse retina. J. Neurosci. 30, 4048–4061 (2010).
Koike, C. et al. TRPM1 is a component of the retinal ON bipolar cell transduction channel in the mGluR6 cascade. Proc. Natl. Acad. Sci. USA 107, 332–337 (2010).
Blackshaw, S. et al. Genomic analysis of mouse retinal development. PLoS Biol. 2, e247 (2004).
He, S. et al. MicroRNA-encoding long non-coding RNAs. BMC Genomics 9, 236 (2008).
Hackler, L., Wan, J., Swaroop, A., Qian, J. & Zack, D.J. MicroRNA profile of the developing mouse retina. Invest. Ophthalmol. Vis. Sci. 51, 1823–1831 (2010).
Ohsawa, R. & Kageyama, R. Regulation of retinal cell fate specification by multiple transcription factors. Brain Res. 1192, 90–98 (2008).
Cepko, C.L., Austin, C.P., Yang, X., Alexiades, M. & Ezzeddine, D. Cell fate determination in the vertebrate retina. Proc. Natl. Acad. Sci. USA 93, 589–595 (1996).
Ng, L. et al. A thyroid hormone receptor that is required for the development of green cone photoreceptors. Nat. Genet. 27, 94–98 (2001).
Furukawa, T., Morrow, E.M. & Cepko, C.L. Crx, a novel otx-like homeobox gene, shows photoreceptor-specific expression and regulates photoreceptor differentiation. Cell 91, 531–541 (1997).
Chen, S. et al. Crx, a novel Otx-like paired-homeodomain protein, binds to and transactivates photoreceptor cell-specific genes. Neuron 19, 1017–1030 (1997).
Nishida, A. et al. Otx2 homeobox gene controls retinal photoreceptor cell fate and pineal gland development. Nat. Neurosci. 6, 1255–1263 (2003).
Silber, J. et al. miR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells. BMC Med. 6, 14 (2008).
Côté, F., Collard, J.F. & Julien, J.P. Progressive neuronopathy in transgenic mice expressing the human neurofilament heavy gene: a mouse model of amyotrophic lateral sclerosis. Cell 73, 35–46 (1993).
Okazaki, M.M., Evenson, D.A. & Nadler, J.V. Hippocampal mossy fiber sprouting and synapse formation after status epilepticus in rats: visualization after retrograde transport of biocytin. J. Comp. Neurol. 352, 515–534 (1995).
Chow, R.L. & Lang, R.A. Early eye development in vertebrates. Annu. Rev. Cell Dev. Biol. 17, 255–296 (2001).
Mangale, V.S. et al. Lhx2 selector activity specifies cortical identity and suppresses hippocampal organizer fate. Science 319, 304–309 (2008).
Furukawa, A., Koike, C., Lippincott, P., Cepko, C.L. & Furukawa, T. The mouse Crx 5′-upstream transgene sequence directs cell-specific and developmentally regulated expression in retinal photoreceptor cells. J. Neurosci. 22, 1640–1647 (2002).
Hoesche, C., Sauerwald, A., Veh, R.W., Krippl, B. & Kilimann, M.W. The 5′-flanking region of the rat synapsin I gene directs neuron-specific and developmentally regulated reporter gene expression in transgenic mice. J. Biol. Chem. 268, 26494–26502 (1993).
Qiu, R. et al. The role of miR-124a in early development of the Xenopus eye. Mech. Dev. 126, 804–816 (2009).
Conaco, C., Otto, S., Han, J.J. & Mandel, G. Reciprocal actions of REST and a microRNA promote neuronal identity. Proc. Natl. Acad. Sci. USA 103, 2422–2427 (2006).
Damiani, D. et al. Dicer inactivation leads to progressive functional and structural degeneration of the mouse retina. J. Neurosci. 28, 4878–4887 (2008).
Rajasethupathy, P. et al. Characterization of small RNAs in aplysia reveals a role for miR-124 in constraining synaptic plasticity through CREB. Neuron 63, 803–817 (2009).
Yoo, A.S., Staahl, B.T., Chen, L. & Crabtree, G.R. MicroRNA-mediated switching of chromatin-remodeling complexes in neural development. Nature 460, 642–646 (2009).
Friedman, R.C., Farh, K.K., Burge, C.B. & Bartel, D.P. Most mammalian mRNAs are conserved targets of microRNAs. Genome Res. 19, 92–105 (2009).
Lewis, B.P., Burge, C.B. & Bartel, D.P. Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120, 15–20 (2005).
Lewis, B.P., Shih, I.H., Jones-Rhoades, M.W., Bartel, D.P. & Burge, C.B. Prediction of mammalian microRNA targets. Cell 115, 787–798 (2003).
Doench, J.G. & Sharp, P.A. Specificity of microRNA target selection in translational repression. Genes Dev. 18, 504–511 (2004).
Liu, K. et al. MiR-124 regulates early neurogenesis in the optic vesicle and forebrain, targeting NeuroD1. Nucleic Acids Res. 39, 2869–2879 (2011).
Porter, F.D. et al. Lhx2, a LIM homeobox gene, is required for eye, forebrain and definitive erythrocyte development. Development 124, 2935–2944 (1997).
Wilson, S.I., Shafer, B., Lee, K.J. & Dodd, J. A molecular program for contralateral trajectory: Rig-1 control by LIM homeodomain transcription factors. Neuron 59, 413–424 (2008).
Holmes, G.L., Sarkisian, M., Ben-Ari, Y. & Chevassus-Au-Louis, N. Mossy fiber sprouting after recurrent seizures during early development in rats. J. Comp. Neurol. 404, 537–553 (1999).
Baulac, S. et al. A novel locus for generalized epilepsy with febrile seizures plus in French families. Arch. Neurol. 65, 943–951 (2008).
Glancy, M. et al. Transmitted duplication of 8p23.1–8p23.2 associated with speech delay, autism and learning difficulties. Eur. J. Hum. Genet. 17, 37–43 (2009).
Koyama, R. & Ikegaya, Y. Mossy fiber sprouting as a potential therapeutic target for epilepsy. Curr. Neurovasc. Res. 1, 3–10 (2004).
Weiler, I.J. et al. Fragile X mental retardation protein is translated near synapses in response to neurotransmitter activation. Proc. Natl. Acad. Sci. USA 94, 5395–5400 (1997).
Edbauer, D. et al. Regulation of synaptic structure and function by FMRP-associated microRNAs miR-125b and miR-132. Neuron 65, 373–384 (2010).
Xu, X.L., Li, Y., Wang, F. & Gao, F.B. The steady-state level of the nervous system–specific microRNA-124a is regulated by dFMR1 in Drosophila. J. Neurosci. 28, 11883–11889 (2008).
Siegel, G. et al. A functional screen implicates microRNA-138-dependent regulation of the depalmitoylation enzyme APT1 in dendritic spine morphogenesis. Nat. Cell Biol. 11, 705–716 (2009).
Sanuki, R., Omori, Y., Koike, C., Sato, S. & Furukawa, T. Panky, a novel photoreceptor-specific ankyrin repeat protein, is a transcriptional cofactor that suppresses CRX-regulated photoreceptor genes. FEBS Lett. 584, 753–758 (2010).
Gray, P.A. et al. Mouse brain organization revealed through direct genome-scale TF expression analysis. Science 306, 2255–2257 (2004).
Babb, T.L., Kupfer, W.R., Pretorius, J.K., Crandall, P.H. & Levesque, M.F. Synaptic reorganization by mossy fibers in human epileptic fascia dentata. Neuroscience 42, 351–363 (1991).
Onishi, A. et al. Pias3-dependent SUMOylation directs rod photoreceptor development. Neuron 61, 234–246 (2009).
Acknowledgements
We thank T. Maniatis for RIPmiR-124a-2, M. Kilimann for Synapsin 1 promoter, Y. Omori, K. Terada, M. Ueno, N. Nagata, K. Aritake, Y. Oishi, T. Hamasaki, and H. Abe for critical comments and technical advice, and A. Tani, M. Kadowaki, Y. Kawakami, A. Ishimaru, H. Tsujii, T. Saioka, K. Sone, H. Abe, and S. Kennedy for technical assistance. This work was supported by Japan Science and Technology Agency (JST), Core Research for Evolutional Science and Technology (CREST), Grant-in-Aid for Scientific Research (B), Grant-in-Aid for Young Scientists (B), a Grant for Molecular Brain Science from the Ministry of Education, Culture, Sports, Science and Technology, the Takeda Science Foundation, the Uehara Memorial Foundation, the Mochida Memorial Foundation, and the Naito Foundation.
Author information
Authors and Affiliations
Contributions
R.S. and T.F. designed the project. R.S., C.K., S.W., S.I. and T.F carried out the molecular and in situ hybridization experiments. R.S. and A.O. performed in vivo electroporation, virus infection and knockdown experiments in retinal and hippocampal neurons, and immunohistochemistry. S.U., T.K., M.K. and R.S. carried out the ERG experiments. R.S., Y.M. and T.F. produced the knockout and transgenic mice. R.S., R. Muramatsu and T.Y. carried out hippocampal tissue experiments. R.S., R. Matsui and D.W. produced lentivirus. R.S., Y.C. and Y.U. produced adeno-associated virus. R.S. and T.F. wrote the manuscript. T.F. supervised the project.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–12, Supplementary Tables 1 and 2, and Supplementary Statistical Analysis (PDF 2567 kb)
Rights and permissions
About this article
Cite this article
Sanuki, R., Onishi, A., Koike, C. et al. miR-124a is required for hippocampal axogenesis and retinal cone survival through Lhx2 suppression. Nat Neurosci 14, 1125–1134 (2011). https://doi.org/10.1038/nn.2897
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nn.2897
This article is cited by
-
MicroRNA‑124: an emerging therapeutic target in central nervous system disorders
Experimental Brain Research (2023)
-
Molecular Mechanisms Involved in the Regulation of Neurodevelopment by miR-124
Molecular Neurobiology (2023)
-
Single-Nucleotide Variants in microRNAs Sequences or in their Target Genes Might Influence the Risk of Epilepsy: A Review
Cellular and Molecular Neurobiology (2022)
-
MicroRNAs and Synaptic Plasticity: From Their Molecular Roles to Response to Therapy
Molecular Neurobiology (2022)
-
Therapeutic effects of mesenchymal stem cells-derived extracellular vesicles’ miRNAs on retinal regeneration: a review
Stem Cell Research & Therapy (2021)