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Neuromuscular synaptic patterning requires the function of skeletal muscle dihydropyridine receptors

Nature Neuroscience volume 14, pages 570–577 (2011)Cite this article

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Abstract

Developing skeletal myofibers in vertebrates are intrinsically 'pre-patterned' for motor nerve innervation. However, the intrinsic factors that regulate muscle pre-patterning remain unknown. We found that a functional skeletal muscle dihydropyridine receptor (DHPR, the L-type Ca2+ channel in muscle) was required for muscle pre-patterning during the development of the neuromuscular junction (NMJ). Targeted deletion of the β1 subunit of DHPR (Cacnb1) in mice led to muscle pre-patterning defects, aberrant innervation and precocious maturation of the NMJ. Reintroducing Cacnb1 into Cacnb1−/− muscles reversed the pre-patterning defects and restored normal development of the NMJ. The mechanism by which DHPRs govern muscle pre-patterning is independent of their role in excitation-contraction coupling, but requires Ca2+ influx through the L-type Ca2+ channel. Our findings indicate that the skeletal muscle DHPR retrogradely regulates the patterning and formation of the NMJ.

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Figure 1: Loss of DHPR function leads to defects in muscle pre-patterning.
Figure 2: Loss of DHPR function leads to multiple synaptic sites and an expansion of innervation territories.
Figure 3: DHPR function is not required for synaptogenesis, but its absence leads to increased synaptic and muscle electrical activity.
Figure 4: Muscle-specific expression of Cacnb1 rescues the patterning defects in Cacnb1−/− muscle.
Figure 5: RyRs and DHPRs have different roles in muscle pre-patterning.
Figure 6: DHPRs pattern neuromuscular synapses by regulating the expression of the AChR genes and Musk.

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Acknowledgements

We thank J. Takahashi, E. Olson, B. Szaro, J. Johnson, C. Green, K. Walton, J. Terman, J. McArdle and R. Bassel-Duby for critical reading and commenting on the manuscript, S. Cannon, D. Francis, E. Kavalali and I. Bezprozvanny for valuable suggestions on muscle physiology and S. Burden for AChRα and MuSK in situ probes. This study was supported by grants (to W.L.) from the US National Institutes of Health/National Institute of Neurological Disorders and Stroke (NS 055028), the Edward Mallinckrodt, Jr. Scholar Program, and the Cain Foundation in Medical Research at University of Texas Southwestern Medical Center, and by a grant (to P.D.A.) from the US National Institutes of Health/National Institute of Arthritis and Musculoskeletal and Skin Diseases (P01AR044750).

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Authors and Affiliations

  1. Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, Texas, USA

    Fujun Chen, Yun Liu, Yoshie Sugiura & Weichun Lin

  2. Department of Anesthesia, Perioperative and Pain Medicine, Brigham and Women's Hospital, Boston, Massachusetts, USA

    Paul D Allen

  3. Department of Biochemistry and Molecular Biology, University of Louisville, Louisville, Kentucky, USA

    Ronald G Gregg

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  1. Fujun Chen
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Contributions

F.C., Y.L. and Y.S. carried out the experiment and collected and analyzed the data. P.D.A. provided the RyR mutant mice. R.G.G. provided the DHPR mutant mice. W.L. supervised the project and wrote the manuscript with critical input from F.C., Y.L., Y.S., P.D.A. and R.G.G.

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Correspondence to Weichun Lin.

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Chen, F., Liu, Y., Sugiura, Y. et al. Neuromuscular synaptic patterning requires the function of skeletal muscle dihydropyridine receptors. Nat Neurosci 14, 570–577 (2011). https://doi.org/10.1038/nn.2792

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