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Balanced gene regulation by an embryonic brain ncRNA is critical for adult hippocampal GABA circuitry

Nature Neuroscience volume 12, pages 1020–1027 (2009)Cite this article

Abstract

Genomic studies demonstrate that, although the majority of the mammalian genome is transcribed, only about 2% of these transcripts are code for proteins. We investigated how the long, polyadenylated Evf2 noncoding RNA regulates transcription of the homeodomain transcription factors DLX5 and DLX6 in the developing mouse forebrain. We found that, in developing ventral forebrain, Evf2 recruited DLX and MECP2 transcription factors to important DNA regulatory elements in the Dlx5/6 intergenic region and controlled Dlx5, Dlx6 and Gad1 expression through trans and cis-acting mechanisms. Evf2 mouse mutants had reduced numbers of GABAergic interneurons in early postnatal hippocampus and dentate gyrus. Although the numbers of GABAergic interneurons and Gad1 RNA levels returned to normal in Evf2 mutant adult hippocampus, reduced synaptic inhibition occurred. These results suggest that noncoding RNA–dependent balanced gene regulation in embryonic brain is critical for proper formation of GABA-dependent neuronal circuitry in adult brain.

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Figure 1: Evf2TS/TS mice have increased Dlx5 and Dlx6 expression in the embryonic brain.
Figure 2: Loss of Evf2 affects DLX and MECP2 binding to Dlx5/6 intergenic enhancers in E13.5 MGE.
Figure 3: Evf2 does not affect DLX or MECP2 nuclear localization.
Figure 4: GABAergic interneuron loss in the P2 hippocampus and dentate gyrus of Evf2TS/TS mutant mice.
Figure 5: Evf2 trans-positively regulates Gad1 expression in E13.5 MGE, but not adult hippocampus.
Figure 6: GABAergic synaptic inhibition is reduced in CA1 layer of the adult hippocampus of Evf2TS/TS mutant mice.

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Acknowledgements

We thank K. Campbell for providing early passage W4 embryonic stem cells, and training and guidance in ES cell manipulation, M. Ekker for Dlx5/6 BAC, P. Lui and N. Copeland for PL253, PL452, bacterial strains and protocols for BAC recombineering, G. Taborn and P. Iannaccone for blastocyst injections, D. Eisentstat for antibody specific to DLX2, A. Joyner for the triple polyadenylation construct, J. Rubenstein for Gad1 probe, and Q. Ma for v-Glut1 probe. We thank K. Jones (Northwestern University) for establishing the conditions for embryonic tissue ChIP. This work was funded by National Institute of Child Health and Human Development grants RO1 HD044745 and R21 HD049875, the Illinois Regenerative Medicine Institute, and an Illinois Excellence in Academic Medicine grant to J.D.K.

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  1. Michael J W VanGompel & Julie C Savage

    Present address: Present address: Division of Reproductive Biology Research, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA (M.J.W.V.); Neurosciences Department, Case Western Reserve University, Cleveland, Ohio, USA (J.C.S.).,

Authors and Affiliations

  1. Developmental Biology and Department of Pediatrics, Children's Memorial Hospital and Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA

    Allison M Bond, Michael J W VanGompel, Mary F Clark, Julie C Savage & Jhumku D Kohtz

  2. Department of Physiology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA

    Evgeny A Sametsky & John F Disterhoft

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Contributions

A.M.B. performed the experiments in Figures 1,2,3,4,5. M.J.W.V. carried out BAC recombineering, ES cell homologous targeting and screening for generating Evf2 TS mice. J.C.S. performed the experiments shown in Figures 1, 3 and 5. M.F.C. carried out the experiments shown in Figures 1, 5 and Supplementary Figure 1. E.A.S. and J.F.D. contributed electrophysiology experiments and results for Figure 6. J.D.K. conceived of and directed experiments and wrote the manuscript.

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Correspondence to Jhumku D Kohtz.

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Bond, A., VanGompel, M., Sametsky, E. et al. Balanced gene regulation by an embryonic brain ncRNA is critical for adult hippocampal GABA circuitry. Nat Neurosci 12, 1020–1027 (2009). https://doi.org/10.1038/nn.2371

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