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A role for mDia, a Rho-regulated actin nucleator, in tangential migration of interneuron precursors

Nature Neuroscience volume 15, pages 373–380 (2012)Cite this article

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Abstract

In brain development, distinct types of migration, radial migration and tangential migration, are shown by excitatory and inhibitory neurons, respectively. Whether these two types of migration operate by similar cellular mechanisms remains unclear. We examined neuronal migration in mice deficient in mDia1 (also known as Diap1) and mDia3 (also known as Diap2), which encode the Rho-regulated actin nucleators mammalian diaphanous homolog 1 (mDia1) and mDia3. mDia deficiency impaired tangential migration of cortical and olfactory inhibitory interneurons, whereas radial migration and consequent layer formation of cortical excitatory neurons were unaffected. mDia-deficient neuroblasts exhibited reduced separation of the centrosome from the nucleus and retarded nuclear translocation. Concomitantly, anterograde F-actin movement and F-actin condensation at the rear, which occur during centrosomal and nuclear movement of wild-type cells, respectively, were impaired in mDia-deficient neuroblasts. Blockade of Rho-associated protein kinase (ROCK), which regulates myosin II, also impaired nuclear translocation. These results suggest that Rho signaling via mDia and ROCK critically regulates nuclear translocation through F-actin dynamics in tangential migration, whereas this mechanism is dispensable in radial migration.

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Figure 1: Radial migration of excitatory cortical precursors is not impaired in mDia DKO mice.
Figure 2: Tangential migration of cortical interneurons is impaired in mDia DKO mice.
Figure 3: Migration of SVZ neuroblasts to the granule cell layer of the olfactory bulb is interrupted in mDia DKO mice.
Figure 4: mDia deficiency causes impaired migration from MGE and SVZ explants in vitro.
Figure 5: Fluorescent imaging analyses of cell morphology and movement of SVZ neuroblasts.
Figure 6: mDia DKO neuroblasts show reduced movement of the centrosome and the cell body.
Figure 7: mDia-dependent F-actin dynamics and localization of mDia in migrating SVZ neuroblasts.
Figure 8: Roles of ROCK and myosin II in migration of SVZ neuroblasts.

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Acknowledgements

We thank M. Okabe for providing pCX-EGFP, F. Matsuzaki for providing pCAG-EGFP-C1 and pCAG-PACT-mKO1, Y. Yanagawa for providing GAD65 and GAD67 antisense probes, Y. Deguchi for providing mDia3 riboprobes, A. Mizutani for animal care, K. Tohyama for supporting genotyping and plasmid construction, T. Arai and A. Washimi for secretarial help, and K. Nonomura for technical assistance. R.S. thanks A. Kakizuka for providing the opportunity to work in Kyoto University Graduate School of Medicine. R.S. was supported by the Japan Society for the Promotion of Science Research Fellowship. This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan and a Core Research for Evolutional Science and Technology (CREST) grant from Japan Science and Technology Agency.

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  1. Keisuke Watanabe & Hirohide Takebayashi

    Present address: Present address: Division of Neurobiology and Anatomy, Graduate School of Medical and Dental Sciences, Niigata University, Niigata, Japan.,

Authors and Affiliations

  1. Department of Pharmacology, Kyoto University Graduate School of Medicine, Kyoto, Japan

    Ryota Shinohara, Dean Thumkeo, Hiroshi Kamijo, Toshimasa Ishizaki, Tomoyuki Furuyashiki & Shuh Narumiya

  2. Department of Developmental and Regenerative Biology, Institute of Molecular Medicine, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan

    Naoko Kaneko & Kazunobu Sawamoto

  3. Department of Morphological Neural Science, Graduate School of Medical Science, Kumamoto University, Kumamoto, Japan

    Keisuke Watanabe & Hirohide Takebayashi

  4. Laboratory for Animal Resources and Genetic Engineering, RIKEN Center for Developmental Biology, Kobe, Japan

    Hiroshi Kiyonari

  5. Core Research for Evolutional Science and Technology, Tokyo, Japan

    Tomoyuki Furuyashiki & Shuh Narumiya

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Contributions

R.S., D.T., T.F. and S.N. designed the study. R.S. performed most of the experiments. H. Kamijo, R.S., H. Kiyonari, T.I. and S.N. generated mDia3 knockout and mDia DKO mice. D.T. and R.S. carried out in utero electroporation for live imaging. R.S. collected and analyzed the imaging data. N.K. and K.S. performed the whole-mount immunostaining. K.W. and H.T. provided essential advices in setting up and performing in utero electroporation, and conducted in situ hybridization. S.N. and T.F. supervised the project. R.S., T.F. and S.N. wrote the manuscript. All authors discussed the results and concurred on the contents of this manuscript.

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Correspondence to Shuh Narumiya.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–15 (PDF 7632 kb)

Supplementary Video 1

Time-lapse, phase-contrast images of neuroblast migration from a wild-type SVZ explant in Matrigel. Images were acquired at 5 min intervals for 8 h. (MOV 2141 kb)

Supplementary Video 2

Time-lapse, phase-contrast images of neuroblast migration from an mDia-DKO SVZ explant in Matrigel. Images were acquired at 5 min intervals for 8 h. (MOV 1343 kb)

Supplementary Video 3

Time-lapse fluorescent images of migrating SVZ neuroblasts expressing EGFP from a wild-type explant in Matrigel. Images were acquired at 3 min intervals for 45 min. (MOV 340 kb)

Supplementary Video 4

Time-lapse fluorescent images of migrating SVZ neuroblasts expressing EGFP from an mDia-DKO explant in Matrigel. Images were acquired at 3 min intervals for 45 min. (MOV 940 kb)

Supplementary Video 5

Time-lapse fluorescent images of a wild-type SVZ neuroblast during the nuclear translocation in Matrigel. Images were acquired at 3 min intervals for 15 min. Signals of EGFP and PACT-mKO1 are shown in green and purple, respectively. (MOV 566 kb)

Supplementary Video 6

Time-lapse fluorescent images of an mDia-DKO SVZ neuroblast during the nuclear translocation in Matrigel. Images were acquired at 3 min intervals for 33 min. Signals of EGFP and PACT-mKO1 are shown in green and purple, respectively. (MOV 783 kb)

Supplementary Video 7

Time-lapse fluorescent images of F-actin dynamics during the nuclear translocation in two representative wild-type SVZ neuroblasts. Images were acquired at 3 min intervals for 21 or 18 min for respective neuroblasts. Signals of Lifeact-EGFP and PACT-mKO1 are shown in green and purple, respectively. (MOV 1459 kb)

Supplementary Video 8

Time-lapse fluorescent images of F-actin dynamics during the nuclear translocation in two representative mDia-DKO SVZ neuroblasts. Images were acquired at 3 min intervals for 15 or 33 min for respective neuroblasts. Signals of Lifeact-EGFP and PACT-mKO1 are shown in green and purple, respectively. (MOV 1285 kb)

Supplementary Video 9

Time-lapse fluorescent images of a migrating SVZ neuroblast without Y-27632, a ROCK inhibitor, in Matrigel. Images were acquired at 3 min intervals for 36 min. Signals of EGFP and PACT-mKO1 are shown in green and purple, respectively. (MOV 1154 kb)

Supplementary Video 10

Time-lapse fluorescent images of a migrating SVZ neuroblast treated with Y-27632, a ROCK inhibitor, in Matrigel. Images were acquired at 3 min intervals for 30 min. Signals of EGFP and PACT-mKO1 are shown in green and purple, respectively. (MOV 2617 kb)

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Shinohara, R., Thumkeo, D., Kamijo, H. et al. A role for mDia, a Rho-regulated actin nucleator, in tangential migration of interneuron precursors. Nat Neurosci 15, 373–380 (2012). https://doi.org/10.1038/nn.3020

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