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Serotonin modulates the response of embryonic thalamocortical axons to netrin-1

Nature Neuroscience volume 10, pages 588–597 (2007)Cite this article

Abstract

Modifying serotonin (5-HT) abundance in the embryonic mouse brain disrupts the precision of sensory maps formed by thalamocortical axons (TCAs), suggesting that 5-HT influences their growth. We investigated the mechanism by which 5-HT influences TCAs during development. 5-HT1B and 5-HT1D receptor expression in the fetal forebrain overlaps with that of the axon guidance receptors DCC and Unc5c. In coculture assays, axons originating from anterior and posterior halves of the embryonic day 14.5 dorsal thalamus responded differently to netrin-1, reflecting the patterns of DCC and Unc5c expression. 5-HT converts the attraction exerted by netrin-1 on posterior TCAs to repulsion. Pharmacological manipulation of 5-HT1B/1D receptors and intracellular cAMP showed the signaling cascade through which this modulation occurs. An in vivo correlate of altered TCA pathfinding was obtained by transient manipulation of 5-HT1B/1D receptor expression abundance in the dorsal thalamus by in utero electroporation. These data demonstrate that serotonergic signaling has a previously unrecognized role in the modulation of axonal responsiveness to a classic guidance cue.

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Figure 1: Bright-field micrographs illustrate the distribution of Htr1b, Htr1d, Dcc and Unc5c receptor transcripts and Gbx2 transcription factor transcripts in embryonic dorsal thalamus (DT).
Figure 2: Analysis of attractive and repulsive effects of netrin-1 and slit-2 on DT axons in vitro.
Figure 3: 5-HT1B/1D receptors mediate 5-HT modulation of DT axons response to netrin-1.
Figure 4: Modulation of the posterior DT axon response to netrin-1 is mediated by a 5-HT1B/1D–mediated change of intracellular cAMP.
Figure 5: In utero electroporation of E12.5 DT neurons to examine TCA pathway formation in vivo.
Figure 6: Modifying 5-HT1B/1D receptor expression in vivo alters the topography of a subpopulation of TCAs.
Figure 7: Ntn1 (netrin-1) transcript expression in the striatum/IC area at E14.5 and E16.5.
Figure 8: Modifying 5-HT1B/1D receptor expression in vivo alters TCA distribution in the internal capsule.

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Acknowledgements

We thank J. Wu and H.H. Wu for technical help and comments, and K.L. Eagleson and G. Stanwood for comments on the manuscript. The work was supported in part by US National Institute of Mental Health grant MH65299 and US National Institute of Child Health and Human Development P30HD15052 to P.L.

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Author notes

  1. Alexandre Bonnin and Masaaki Torii: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmacology, Vanderbilt University, 465 21st Ave. South, Nashville, 37232, Tennessee, USA

    Alexandre Bonnin, Masaaki Torii & Pat Levitt

  2. Vanderbilt Kennedy Center for Research on Human Development, Vanderbilt University, Box 40 Peabody, 230 Appleton Place, Nashville, 37203, Tennessee, USA

    Alexandre Bonnin, Masaaki Torii & Pat Levitt

  3. Department of Neurobiology and Kavli Institute of Neuroscience, Yale University Medical School, 333 Cedar St SHM C316, New Haven, 06510, Connecticut, USA

    Masaaki Torii & Pasko Rakic

  4. Department of Biostatistics, Vanderbilt University School of Medicine, S-2323 Medical Center North, Nashville, 37232, Tennessee, USA

    Lilly Wang

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Contributions

A.B. participated in the design of the project, the writing of the manuscript and conducted and analyzed the experiments. M.T. conducted and participated in the analyses of crucial in utero electroporation experiments and helped to write the manuscript. L.W. performed the statistical analyses. P.R. contributed to the in vivo experiments and helped to write the manuscript. P.L. contributed to the design of the experiments, interpretation of results and writing of the manuscript.

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Correspondence to Pat Levitt.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Putative 5-HT signaling in development. (PDF 496 kb)

Supplementary Fig. 2

Putative 5-HT signaling in development. (PDF 892 kb)

Supplementary Fig. 3

Quantification of netrin-G1 immunofluorescent intensity at E18.5 on the control and electroporated side of brains electroporated at E12.5. (PDF 170 kb)

Supplementary Fig. 4

Pathways of TCAs from anterior and posterior DT in horizontal sections at E14.5. (PDF 1312 kb)

Supplementary Fig. 5

Anterior-posterior positioning of TCAs in the IC is associated with their medial-lateral, rather than anterior-posterior, origin in the DT. (PDF 1262 kb)

Supplementary Fig. 6

Comparison between Gbx2 and Unc5c expression patterns in dorso-ventral series of horizontal sections through the E14.5 DT. (PDF 489 kb)

Supplementary Fig. 7

Schematic representation of TCA pathfinding. (PDF 63 kb)

Supplementary Methods (PDF 115 kb)

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Bonnin, A., Torii, M., Wang, L. et al. Serotonin modulates the response of embryonic thalamocortical axons to netrin-1. Nat Neurosci 10, 588–597 (2007). https://doi.org/10.1038/nn1896

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