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GABAergic excitation after febrile seizures induces ectopic granule cells and adult epilepsy

Nature Medicine volume 18, pages 1271–1278 (2012)Cite this article

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Abstract

Temporal lobe epilepsy (TLE) is accompanied by an abnormal location of granule cells in the dentate gyrus. Using a rat model of complex febrile seizures, which are thought to be a precipitating insult of TLE later in life, we report that aberrant migration of neonatal-generated granule cells results in granule cell ectopia that persists into adulthood. Febrile seizures induced an upregulation of GABAA receptors (GABAA-Rs) in neonatally generated granule cells, and hyperactivation of excitatory GABAA-Rs caused a reversal in the direction of granule cell migration. This abnormal migration was prevented by RNAi-mediated knockdown of the Na+K+2Cl− co-transporter (NKCC1), which regulates the excitatory action of GABA. NKCC1 inhibition with bumetanide after febrile seizures rescued the granule cell ectopia, susceptibility to limbic seizures and development of epilepsy. Thus, this work identifies a previously unknown pathogenic role of excitatory GABAA-R signaling and highlights NKCC1 as a potential therapeutic target for preventing granule cell ectopia and the development of epilepsy after febrile seizures.

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Figure 1: Febrile seizures induce ectopic granule cells.
Figure 2: GABAA-R signaling modulates granule cell migration.
Figure 3: GABAA-R hyperactivity underlies aberrant granule cell migration.
Figure 4: NKCC1 inhibition rescues granule cell migration deficits and ectopia.
Figure 5: Bumetanide rescues granule cell ectopia, seizure susceptibility and development of epilepsy.
Figure 6: Bumetanide blocks the emergence of ectopic granule cells and epileptic ripples in adulthood.

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Acknowledgements

This work was supported by a Grant-in-Aid for Science Research on Young Scientists (B) (19790048) and on Innovative Areas 'Mesoscopic Neurocircuitry' (22115003) from The Ministry of Education, Culture, Sports, Science and Technology of Japan; the Research Foundation for Pharmaceutical Sciences; and the Funding Program for Next Generation World-Leading Researchers (LS023).

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  1. Kentaro Tao and Takuya Sasaki: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, Japan

    Ryuta Koyama, Kentaro Tao, Takuya Sasaki, Junya Ichikawa, Daisuke Miyamoto, Rieko Muramatsu, Norio Matsuki & Yuji Ikegaya

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Contributions

R.K. designed the project, conducted and analyzed the experiments, and wrote the manuscript. K.T. did in vivo electrophysiology, long-term video monitoring and helped with preparation of retroviruses. T.S. did in vitro electrophysiology and time-lapse imaging in explant cultures. J.I. did injection of retroviruses. D.M. helped with in vivo electrophysiology. R.M. helped with perfusion of mice. N.M. and Y.I. are senior authors and were responsible for project planning. All authors analyzed and discussed the results and commented on the manuscript.

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Correspondence to Ryuta Koyama.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Methods and Supplementary Figures 1–9 (PDF 1062 kb)

Supplementary Video 1

Radial migration of a GFP+ GC (Supplementary Fig. 4a). The images were acquired every 1 h for 24 h during the 3 to 4 DIV. (MOV 349 kb)

Supplementary Video 2

Somal translocation of a GFP+ GC (Supplementary Fig. 4b). The images were acquired every 1 h for 24 h during the 3 to 4 DIV. (MOV 343 kb)

Supplementary Video 3

Aberrant migration of a GC from a HT rat in Figure 3c. The images were acquired every 1 h for 6 h beginning at 3 DIV. (MOV 92 kb)

Supplementary Video 4

A migrating GC from the explant culture prepared from a NT rat (Fig. 4b, left). The images were acquired at 1 frame per 2 min for 60 min. Muscimol was focally applied to the leading growth cone for 1 min at the time point of 0 min. (MOV 545 kb)

Supplementary Video 5

A migrating GC from the explant culture prepared from a HT rat (Fig. 4b, right). The images were acquired at 1 frame per 2 min for 60 min. Muscimol was focally applied to the leading growth cone for 1 min at the time point of 0 min. (MOV 261 kb)

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Koyama, R., Tao, K., Sasaki, T. et al. GABAergic excitation after febrile seizures induces ectopic granule cells and adult epilepsy. Nat Med 18, 1271–1278 (2012). https://doi.org/10.1038/nm.2850

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