Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Dual-mode operation of neuronal networks involved in left–right alternation

Abstract

All forms of locomotion are repetitive motor activities that require coordinated bilateral activation of muscles. The executive elements of locomotor control are networks of spinal neurons that determine gait pattern through the sequential activation of motor-neuron pools on either side of the body axis1,2,3,4. However, little is known about the constraints that link left–right coordination to locomotor speed. Recent advances have indicated that both excitatory and inhibitory commissural neurons may be involved in left–right coordination5,6,7. But the neural underpinnings of this, and a possible causal link between these different groups of commissural neurons and left–right alternation, are lacking. Here we show, using intersectional mouse genetics, that ablation of a group of transcriptionally defined commissural neurons—the V0 population—leads to a quadrupedal hopping at all frequencies of locomotion. The selective ablation of inhibitory V0 neurons leads to a lack of left–right pattern at low frequencies, mixed coordination at medium frequencies, and alternation at high locomotor frequencies. When ablation is targeted to excitatory V0 neurons, left–right alternation is present at low frequencies, and hopping is restricted to medium and high locomotor frequencies. Therefore, the intrinsic logic of the central control of locomotion incorporates a modular organization, with two subgroups of V0 neurons required for the existence of left–right alternating modes at different speeds of locomotion. The two molecularly distinct sets of commissural neurons may constrain species-related naturally occurring frequency-dependent coordination and be involved in the evolution of different gaits.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Ablation of V0 neurons leads to a hopping gait.
Figure 2: V0-deleted mice hop at all locomotor frequencies.
Figure 3: V0D-deleted mice show a lack of alternation at low frequencies and alternation at high frequencies.
Figure 4: Deletion of excitatory V0V neurons causes hopping at high locomotor frequencies but preserves alternation at lower frequencies.

References

  1. Grillner, S. The motor infrastructure: from ion channels to neuronal networks. Nature Rev. Neurosci. 4, 573–586 (2003)

    CAS  Article  Google Scholar 

  2. Kiehn, O. Locomotor circuits in the mammalian spinal cord. Annu. Rev. Neurosci. 29, 279–306 (2006)

    CAS  Article  Google Scholar 

  3. Goulding, M. Circuits controlling vertebrate locomotion: moving in a new direction. Nature Rev. Neurosci. 10, 507–518 (2009)

    CAS  Article  Google Scholar 

  4. Grillner, S. & Jessell, T. M. Measured motion: searching for simplicity in spinal locomotor networks. Curr. Opin. Neurobiol. 19, 572–586 (2009)

    CAS  Article  Google Scholar 

  5. Quinlan, K. A. & Kiehn, O. Segmental, synaptic actions of commissural interneurons in the mouse spinal cord. J. Neurosci. 27, 6521–6530 (2007)

    CAS  Article  Google Scholar 

  6. Jankowska, E. Spinal interneuronal networks in the cat: elementary components. Brain Res. Rev. 57, 46–55 (2008)

    Article  Google Scholar 

  7. Butt, S. J. & Kiehn, O. Functional identification of interneurons responsible for left-right coordination of hindlimbs in mammals. Neuron 38, 953–963 (2003)

    CAS  Article  Google Scholar 

  8. Moran-Rivard, L. et al. Evx1 is a postmitotic determinant of v0 interneuron identity in the spinal cord. Neuron 29, 385–399 (2001)

    CAS  Article  Google Scholar 

  9. Pierani, A. et al. Control of interneuron fate in the developing spinal cord by the progenitor homeodomain protein Dbx1. Neuron 29, 367–384 (2001)

    CAS  Article  Google Scholar 

  10. Lanuza, G. M., Gosgnach, S., Pierani, A., Jessell, T. M. & Goulding, M. Genetic identification of spinal interneurons that coordinate left-right locomotor activity necessary for walking movements. Neuron 42, 375–386 (2004)

    CAS  Article  Google Scholar 

  11. Bielle, F. et al. Multiple origins of Cajal-Retzius cells at the borders of the developing pallium. Nature Neurosci. 8, 1002–1012 (2005)

    CAS  Article  Google Scholar 

  12. Bouvier, J. et al. Hindbrain interneurons and axon guidance signaling critical for breathing. Nature Neurosci. 13, 1066–1074 (2010)

    CAS  Article  Google Scholar 

  13. Berger, J. et al. E1-Ngn2/Cre is a new line for regional activation of cre recombinase in the developing CNS. Genesis 40, 195–199 (2004)

    CAS  Article  Google Scholar 

  14. Teissier, A. et al. A novel transient glutamatergic population migrating from the pallial–subpallial boundary contributes to neocortical development. J. Neurosci. 30, 10563–10574 (2010)

    CAS  Article  Google Scholar 

  15. Witschi, R. et al. Hoxb8-Cre mice: a tool for brain-sparing conditional gene deletion. Genesis 48, 596–602 (2010)

    CAS  Article  Google Scholar 

  16. Serradj, N. & Jamon, M. The adaptation of limb kinematics to increasing walking speeds in freely moving mice 129/Sv and C57BL/6. Behav. Brain Res. 201, 59–65 (2009)

    Article  Google Scholar 

  17. Borgius, L., Restrepo, C. E., Leao, R. N., Saleh, N. & Kiehn, O. A transgenic mouse line for molecular genetic analysis of excitatory glutamatergic neurons. Mol. Cell. Neurosci. 45, 245–257 (2010)

    CAS  Article  Google Scholar 

  18. Keller, C., Hansen, M. S., Coffin, C. M. & Capecchi, M. R. Pax3:Fkhr interferes with embryonic Pax3 and Pax7 function: implications for alveolar rhabdomyosarcoma cell of origin. Genes Dev. 18, 2608–2613 (2004)

    CAS  Article  Google Scholar 

  19. Kjaerulff, O. & Kiehn, O. Distribution of networks generating and coordinating locomotor activity in the neonatal rat spinal cord in vitro: a lesion study. J. Neurosci. 16, 5777–5794 (1996)

    CAS  Article  Google Scholar 

  20. Zagoraiou, L. et al. A cluster of cholinergic premotor interneurons modulates mouse locomotor activity. Neuron 64, 645–662 (2009)

    CAS  Article  Google Scholar 

  21. Zhang, Y. et al. V3 spinal neurons establish a robust and balanced locomotor rhythm during walking. Neuron 60, 84–96 (2008)

    CAS  Article  Google Scholar 

  22. Andersson, L. S. et al. Mutations in DMRT3 affect locomotion in horses and spinal circuit function in mice. Nature 488, 642–646 (2012)

    CAS  ADS  Article  Google Scholar 

  23. Roberts, A., Li, W. C., Soffe, S. R. & Wolf, E. Origin of excitatory drive to a spinal locomotor network. Brain Res. Rev. 57, 22–28 (2008)

    Article  Google Scholar 

  24. McLean, D. L. & Fetcho, J. R. Spinal interneurons differentiate sequentially from those driving the fastest swimming movements in larval zebrafish to those driving the slowest ones. J. Neurosci. 29, 13566–13577 (2009)

    CAS  Article  Google Scholar 

  25. Ausborn, J., Mahmood, R. & El Manira, A. Decoding the rules of recruitment of excitatory interneurons in the adult zebrafish locomotor network. Proc. Natl Acad. Sci. USA 109, E3631–E3639 (2012)

    CAS  ADS  Article  Google Scholar 

  26. Crone, S. A. et al. Genetic ablation of V2a ipsilateral interneurons disrupts left-right locomotor coordination in mammalian spinal cord. Neuron 60, 70–83 (2008)

    CAS  Article  Google Scholar 

  27. Crone, S. A., Zhong, G., Harris-Warrick, R. & Sharma, K. In mice lacking V2a interneurons, gait depends on speed of locomotion. J. Neurosci. 29, 7098–7109 (2009)

    CAS  Article  Google Scholar 

  28. Jordan, L. in Neurobiological Basis of Human Locomotion (ed. Shimamura, M. ) 3–21 (Japan Scientific Societies, 1991)

    Google Scholar 

  29. Suster, M. L. et al. A novel conserved evx1 enhancer links spinal interneuron morphology and cis-regulation from fish to mammals. Dev. Biol. 325, 422–433 (2009)

    CAS  Article  Google Scholar 

  30. Liu, B. et al. Selective expression of Bhlhb5 in subsets of early-born interneurons and late-born association neurons in the spinal cord. Dev. Dyn. 236, 829–835 (2007)

    CAS  Article  Google Scholar 

  31. Zeilhofer, H. U. et al. Glycinergic neurons expressing enhanced green fluorescent protein in bacterial artificial chromosome transgenic mice. J. Comp. Neurol. 482, 123–141 (2005)

    CAS  Article  Google Scholar 

  32. Restrepo, C. E. et al. Transmitter-phenotypes of commissural interneurons in the lumbar spinal cord of newborn mice. J. Comp. Neurol. 517, 177–192 (2009)

    CAS  Article  Google Scholar 

  33. Tamamaki, N. et al. Green fluorescent protein expression and colocalization with calretinin, parvalbumin, and somatostatin in the GAD67-GFP knock-in mouse. J. Comp. Neurol. 467, 60–79 (2003)

    CAS  Article  Google Scholar 

  34. Talpalar, A. E. & Kiehn, O. Glutamatergic mechanisms for speed control and network operation in the rodent locomotor CPG. Front Neural Circuits 4, 1–14 (2010)

    Google Scholar 

  35. Zar, J. H. Biostatistical Analysis (Prentice-Hall, 1974)

    Google Scholar 

  36. Mor, Y. & Lev-Tov, A. Analysis of rhythmic patterns produced by spinal neural networks. J. Neurophysiol. 98, 2807–2817 (2007)

    CAS  Article  Google Scholar 

  37. Mahan, R. P. Circular Statistical Methods: Applications in Spatial and Temporal Performance Analysis. Special Research Report #16. (United States Army Research Institute for the Behavioral and Social Sciences, ARI Press, 1991)

  38. Skaggs, K., Martin, D. M. & Novitch, B. G. Regulation of spinal interneuron development by the Olig-related protein Bhlhb5 and Notch signaling. Development 138, 3199–3211 (2011)

    CAS  Article  Google Scholar 

  39. Müller, T. et al. The homeodomain factor lbx1 distinguishes two major programs of neuronal differentiation in the dorsal spinal cord. Neuron 34, 551–562 (2002)

    Article  Google Scholar 

Download references

Acknowledgements

This work was supported by the Söderberg Foundation, Swedish Research Council and European Research Council. J.B. is an EMBO fellow. We thank H. U. Zeilhofer for donating the Hoxb8::Cre mouse strain, S. Karaz, A. C. Westerdahl and P. Löw for extensive genotyping, L. Lundfald for participating in early experiments, N. Sleiers for participating in the in vivo experiments, and P. L. Ruffault for providing mouse tissue. We thank colleagues for discussing different aspects of our study.

Author information

Authors and Affiliations

Authors

Contributions

O.K., A.E.T., L.B. and J.B. contributed to the conception and design of the study. A.E.T. performed electrophysiological experiments, J.B. performed anatomical experiments and both analysed the data. L.B. carried out and analysed the in vivo experiments. A.P. engineered the Dbx1DTA mice and detected the hopping phenotype in the E1Ngn2::Cre;Dbx1DTA mice. G.F. provided mice and fixed tissue. O.K. supervised all aspects of the work. All authors discussed the results and participated in writing the manuscript.

Corresponding author

Correspondence to Ole Kiehn.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1-7. (PDF 15651 kb)

Quadrupedal hopping in E1Ngn2::Cre;Dbx1DTA mouse

Three weeks old E1Ngn2::Cre;Dbx1DTA mouse running spontaneously on runway. The run was captured at 100 frames/s and is shown at half‐speed. At all frequencies of locomotion the mouse shows a pronounced quadrupedal hopping with sequential synchronized lifting of the forelimbs followed by synchronized lifting of the hindlimbs. (MP4 9969 kb)

Alternating gait in wild-type mouse

Three weeks old wild‐type mouse running spontaneously on a runway. The run was captured at 100 frames/s and is shown at half‐speed. At frequencies of locomotion below 10 Hz, the mouse produces alternating gaits in forelimbs and hindlimbs. (MP4 7525 kb)

Hindlimb hopping in Hoxb8::Cre; DbxDTA mouse

Three weeks old Hoxb8::Cre; DbxDTA mouse running spontaneously on a runway. The run was captured at 100 frames/s and is shown at half‐speed. At all frequencies of locomotion the mouse displays hindlimb hopping while the forelimb are mostly alternating reflecting the rostro‐caudal gradient of Hoxb8 expression in the spinal cord. (MP4 9328 kb)

Speed dependent hopping in Vglut2::Cre; Dbx1DTA mouse

Three weeks old Vglut2::Cre; Dbx1DTA mouse running spontaneously on a runway. The run was captured at 100 frames/s and is shown at half‐speed. The mouse displays left‐right alternating or hopping hindlimb gait in a frequency‐dependent manner: an alternating pattern is present at low locomotor frequencies (2‐4 Hz) and a hopping gait at high frequencies (4‐10 Hz). Forelimbs are alternating at all frequencies of locomotion. (MP4 8464 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Talpalar, A., Bouvier, J., Borgius, L. et al. Dual-mode operation of neuronal networks involved in left–right alternation. Nature 500, 85–88 (2013). https://doi.org/10.1038/nature12286

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature12286

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing