Specific effects of neuregulin-1β on the communication between DRG neurons and skeletal muscle cells in vitro

  • Menglin Cong
  • Jianmin Li
  • Yuan Qiao
  • Rui Jing
  • Hao Li
  • Zhenzhong LiEmail author


The communication between primary afferent neuron and skeletal muscle (SKM) is one of the important factors on maintaining the structure and function of SKM cells. Neuregulin-1β (NRG-1β) signaling is essential for regulating synaptic neurotransmission. Here, we established a neuromuscular coculture model of dorsal root ganglion (DRG) sensory neurons and SKM cells to explore the nerve-muscle communication in the presence of exogenous NRG-1β. The expression of three distinct subtypes (TrkA, TrkB, and TrkC) of tyrosine kinase receptors was monitored for the phenotypical alterations of the neurons. The aggregation extent of acetylcholine receptor (AChR) represents the specific changes of SKM cells after NRG-1β incubation in this neuromuscular coculture model. The results showed that NRG-1β not only enhanced neurite outgrowth of DRG neurons but also increased the length and branches of SKM cells. NRG-1β treatment not only induced expression of all the three subtypes of Trk receptors in neurons but also promoted AChR aggregation on the surface of SKM cells. The effects of NRG-1β could be blocked by administration of ERK1/2 inhibitor PD98059, PI3K inhibitor LY294002, and JAK2 inhibitor AG490, respectively. These data imply that NRG-1β is essential for the nerve-muscle communication by enhancing growth and modifying phenotypes of the two different kinds of cells. The specific effects produced by NRG-1β add novel interpretation for nerve-muscle communication between sensory neurons and SKM cells.


Tyrosine kinase receptor Neuregulin-1β Signaling pathway Neuron Dorsal root ganglion Skeletal muscle cell 



This work was supported by National Natural Science Foundation of China (No. 81371917).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no potential conflict of interest.


  1. Audisio C, Mantovani C, Raimondo S, Geuna S, Perroteau I, Terenghi G (2012) Neuregulin1 administration increases axonal elongation in dissociated primary sensory neuron cultures. Exp Cell Res 5:570–577CrossRefGoogle Scholar
  2. Basu S, Sladecek S, Pemble H, Wittmann T, Slotman JA, van Cappellen W, Brenner HR, Galjart N (2014) Acetylcholine receptor (AChR) clustering is regulated both by glycogen synthase kinase 3β (GSK3β)-dependent phosphorylation and the level of CLIP-associated protein 2 (CLASP2) mediating the capture of microtubule plus-ends. J Biol Chem 44:30857–30867CrossRefGoogle Scholar
  3. Bucci C, Alifano P, Cogli L (2014) The role of rab proteins in neuronal cells and in the trafficking of neurotrophin receptors. Membranes 4:642–677CrossRefGoogle Scholar
  4. Chang HM, Shyu MK, Tseng GF, Liu CH, Chang HS, Lan CT, Hsu WM, Liao WC (2013) Neuregulin facilitates nerve regeneration by speeding Schwann cell migration via ErbB2/3-dependent FAK pathway. PLoS One 1:e53444CrossRefGoogle Scholar
  5. Dutton EK, Uhm CS, Samuelsson SJ, Schaffner AE, Fitzgerald SC, Daniels MP (1995) Acetylcholine receptor aggregation at nerve-muscle contacts in mammalian cultures: induction by ventral spinal cord neurons is specific to axons. J Neurosci 11:7401–7416CrossRefGoogle Scholar
  6. English AW, Liu K, Nicolini JM, Mulligan AM, Ye K (2013) Small-molecule trkB agonists promote axon regeneration in cut peripheral nerves. Proc Natl Acad Sci USA 40:16217–16222CrossRefGoogle Scholar
  7. Ferraro E, Molinari F, Berghella L (2012) Molecular control of neuromuscular junction development. J Cachexia Sarcopenia Muscle 1:13–23CrossRefGoogle Scholar
  8. Fukazawa T, Matsumoto M, Imura T, Khalesi E, Kajiume T, Kawahara Y, Tanimoto K, Yuge L (2013) Electrical stimulation accelerates neuromuscular junction formation through ADAM19/neuregulin/ErbB signaling in vitro. Neurosci Lett 545:29–34CrossRefGoogle Scholar
  9. Gorokhova S, Gaillard S, Urien L, Malapert P, Legha W, Baronian G, Desvignes JP, Alonso S, Moqrich A (2014) Uncoupling of molecular maturation from peripheral target innervation in nociceptors expressing a chimeric TrkA/TrkC receptor. PLoS Genet 2:e1004081CrossRefGoogle Scholar
  10. Handayaningsih AE, Iguchi G, Fukuoka H, Nishizawa H, Takahashi M, Yamamoto M, Herningtyas EH, Okimura Y, Kaji H, Chihara K, Seino S, Takahashi Y (2011) Reactive oxygen species play an essential role in IGF-I signaling and IGF-I-induced myocyte hypertrophy in C2C12 myocytes. Endocrinology 3:912–921CrossRefGoogle Scholar
  11. Herndon CA, Ankenbruck N, Lester B, Bailey J, Fromm L (2013) Neuregulin1 signaling targets SRF and CREB and activates the muscle spindle-specific gene Egr3 through a composite SRF-CREB-binding site. Exp Cell Res 5:718–730CrossRefGoogle Scholar
  12. Herndon CA, Ankenbruck N, Fromm L (2014) The Erk MAP kinase pathway is activated at muscle spindles and is required for induction of the muscle spindle-specific gene Egr3 by neuregulin1. J Neurosci Res 2:174–184CrossRefGoogle Scholar
  13. Huang EJ, Reichardt LF (2003) Trk receptors: roles in neuronal signal transduction. Annu Rev Biochem 72:609–642CrossRefGoogle Scholar
  14. Ikeda T, Ichii O, Otsuka-Kanazawa S, Nakamura T, Elewa YH, Kon Y (2016) Degenerative and regenerative features of myofibers differ among skeletal muscles in a murine model of muscular dystrophy. J Muscle Res Cell Motil 4–5:153–164CrossRefGoogle Scholar
  15. Jacob J, Tiveron MC, Brunet JF, Guthrie S (2000) Role of the target in the pathfinding of facial visceral motor axons. Mol Cell Neurosci 1:14–26CrossRefGoogle Scholar
  16. Jaworski A, Burden SJ (2006) Neuromuscular synapse formation in mice lacking motor neuron- and skeletal muscle-derived neuregulin-1. J Neurosci 2:655–661CrossRefGoogle Scholar
  17. Kummer TT, Misgeld T, Sanes JR (2006) Assembly of the postsynaptic membrane at the neuromuscular junction: paradigm lost. Curr Opin Neurobiol 1:74–82CrossRefGoogle Scholar
  18. Lai KO, Chen Y, Po HM, Lok KC, Gong K, Ip NY (2004) Identification of the Jak/Stat proteins as novel downstream targets of EphA4 signaling in muscle: implications in the regulation of acetylcholinesterase expression. J Biol Chem 14:13383–13392CrossRefGoogle Scholar
  19. Ledonne A, Nobili A, Latagliata EC, Cavallucci V, Guatteo E (2015) Neuregulin 1 signalling modulates mGluR1 function in mesencephalic dopaminergic neurons. Mol Psychiatry 8:959–973CrossRefGoogle Scholar
  20. Li H, Zhang W, Liu G, Li J, Liu H, Li Z (2012) Expression of tyrosine kinase receptors in cultured dorsal root ganglion neurons in the presence of monosialoganglioside and skeletal muscle cells. J Muscle Res Cell Motil 5:341–350CrossRefGoogle Scholar
  21. Lin W, Burgess RW, Dominguez B, Pfaff SL, Sanes JR, Lee KF (2001) Distinct roles of nerve and muscle in postsynaptic differentiation of the neuromuscular synapse. Nature 6832:1057–1064CrossRefGoogle Scholar
  22. Lu J, Zhou XF, Rush RA (2001) Small primary sensory neurons innervating epidermis and viscera display differential phenotype in the adult rat. Neurosci Res 4:355–363CrossRefGoogle Scholar
  23. Mencel M, Nash M, Jacobson C (2013) Neuregulin upregulates microglial α7 nicotinic acetylcholine receptor expression in immortalized cell lines: implications for regulating neuroinflammation. PLoS One 7:e70338CrossRefGoogle Scholar
  24. Morano M, Ronchi G, Nicolò V, Fornasari BE, Crosio A, Perroteau I, Geuna S, Gambarotta G, Raimondo S (2018) Modulation of the Neuregulin 1/ErbB system after skeletal muscle denervation and reinnervation. Sci Rep 1:5047CrossRefGoogle Scholar
  25. Ngo ST, Cole RN, Sunn N, Phillips WD, Noakes PG (2012) Neuregulin-1 potentiates agrin-induced acetylcholine receptor clustering through muscle-specific kinase phosphorylation. J Cell Sci 125(Pt 6):1531–1543CrossRefGoogle Scholar
  26. Sartini S, Bartolini F, Ambrogini P, Betti M, Ciuffoli S, Lattanzi D, Di Palma M, Cuppini R (2013) Motor activity affects adult skeletal muscle re-innervation acting via tyrosine kinase receptors. Eur J Neurosci 9:1394–1403CrossRefGoogle Scholar
  27. Song F, Chiang P, Wang J, Ravits J, Loeb JA (2012) Aberrant neuregulin 1 signaling in amyotrophic lateral sclerosis. J Neuropathol Exp Neurol 2:104–115CrossRefGoogle Scholar
  28. Wang L, Liu Z, Liu H, Wan Y, Wang H, Li Z (2009) Neuronal phenotype and tyrosine kinase receptor expression in cocultures of dorsal root ganglion and skeletal muscle cells. Anat Rec 1:107–112CrossRefGoogle Scholar
  29. Wang J, Song F, Loeb JA (2017) Neuregulin1 fine-tunes pre-, post-, and perisynaptic neuromuscular junction development. Dev Dyn 246:368–380CrossRefGoogle Scholar
  30. Xie F, Zhang F, Min S, Chen J, Yang J, Wang X (2018) Glial cell line-derived neurotrophic factor (GDNF) attenuates the peripheral neuromuscular dysfunction without inhibiting the activation of spinal microglia/monocyte. BMC Geriatr 18:110CrossRefGoogle Scholar
  31. Yang X, Arber S, William C, Li L, Tanabe Y, Jessell TM, Birchmeier C (2001) Patterning of muscle acetylcholine receptor gene expression in the absence of motor innervation. Neuron 2:399–410CrossRefGoogle Scholar
  32. Yue W, Song L, Fu G, Li Y, Liu H (2013) Neuregulin-1β regulates tyrosine kinase receptor expression in cultured dorsal root ganglion neurons with excitotoxicity induced by glutamate. Regul Pept 180:33–42CrossRefGoogle Scholar
  33. Zhang W, Li Z (2013) The effects of target skeletal muscle cells on dorsal root ganglion neuronal outgrowth and migration in vitro. PLoS One 1:e52849CrossRefGoogle Scholar
  34. Zhang W, Li H, Xing Z, Yuan H, Kindy MS, Li Z (2013a) Expression of mRNAs for PPT, CGRP, NF-200, and MAP-2 in cocultures of dissociated DRG neurons and skeletal muscle cells in administration of NGF or NT-3. Folia Histochem Cytobiol 2:15786Google Scholar
  35. Zhang W, Miao Y, Xing Z, Li H, Liu H, Li Z (2013b) Growth-associated protein-43 expression in cocultures of dorsal root ganglion neurons and skeletal muscle cells with different neurotrophins. Muscle Nerve 6:909–915CrossRefGoogle Scholar
  36. Zhang Y, Lin S, Karakatsani A, Rüegg MA, Kröger S (2015) Differential regulation of AChR clustering in the polar and equatorial region of murine muscle spindles. Eur J Neurosci 1:69–78CrossRefGoogle Scholar

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© Springer Nature Switzerland AG 2018

Authors and Affiliations

  1. 1.Department of AnatomyShandong University School of Basic Medical SciencesJinanChina
  2. 2.Department of OrthopaedicsShandong University Qilu HospitalJinanChina
  3. 3.Medical Imaging Centerthe Second Hospital of Shandong UniversityJinanChina

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