Download PDF
J Reconstr Microsurg 2018; 34(05): 348-358
DOI: 10.1055/s-0038-1627463
Original Article
Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Immunohistochemical Detection of Motor Endplates in the Long-Term Denervated Muscle

Liancai Mu
1   Department of Biomedical Research, Hackensack University Medical Center, Hackensack, New Jersey
,
Jingming Chen
1   Department of Biomedical Research, Hackensack University Medical Center, Hackensack, New Jersey
,
Jing Li
1   Department of Biomedical Research, Hackensack University Medical Center, Hackensack, New Jersey
,
Themba Nyirenda
1   Department of Biomedical Research, Hackensack University Medical Center, Hackensack, New Jersey
,
Mary Fowkes
2   Department of Pathology, Icahn School of Medicine at Mount Sinai, New York, New York
,
Stanislaw Sobotka
1   Department of Biomedical Research, Hackensack University Medical Center, Hackensack, New Jersey
3   Department of Neurosurgery, Icahn School of Medicine at Mount Sinai, New York, New York
› Author Affiliations
Further Information

Publication History

01 September 2017

23 December 2017

Publication Date:
06 March 2018 (online)

Also available at
    Permissions and Reprints

Abstract

Background We have demonstrated that the native motor zone (NMZ) within a muscle is an ideal target for performing nerve-muscle-endplate band grafting (NMEG) to restore motor function of a denervated muscle. This study was designed to determine spatiotemporal alterations of the myofibers, motor endplates (MEPs), and axons in the NMZ of long-term denervated muscles for exploring if NMEG-NMZ technique would have the potential for delayed reinnervation.

Methods Sternomastoid (SM) muscles of adult female Sprague-Dawley rats (n = 21) were experimentally denervated and denervation-induced changes in muscle weight, myofiber size, MEPs, and intramuscular nerve axons were evaluated histomorphometrically and immunohistochemically at the end of 3, 6, and 9 months after denervation. The values obtained from the ipsilateral normal side served as control.

Results The denervated SM muscles exhibited a progressive reduction in muscle weight (38%, 31%, and 19% of the control) and fiber diameter (52%, 40%, and 28% of the control) for 3-, 6-, and 9-month denervation, respectively. The denervated MEPs were still detectable even 9 months after denervation. The mean number of the denervated MEPs was 79%, 65%, and 43% of the control in the 3-, 6-, and 9-month denervated SM, respectively. Degenerated axons in the denervated muscles became fragmented.

Conclusions Persistence of MEPs in the long-term denervated SM suggests that some surgeries targeting the MEPs such as NMEG-NMZ technique should be effective for delayed reinnervation. However, more work is needed to develop strategies for preservation of muscle mass and MEPs after denervation.

Keywords

denervation - peripheral nerve injury - motor endplate - acetylcholine receptor - muscle atrophy - native motor zone
 

  • References

  • 1 Fu SY, Gordon T. Contributing factors to poor functional recovery after delayed nerve repair: prolonged denervation. J Neurosci 1995; 15 (5 Pt 2): 3886-3895
    CrossrefPubMedGoogle Scholar
  • 2 Gordon T, Tyreman N, Raji MA. The basis for diminished functional recovery after delayed peripheral nerve repair. J Neurosci 2011; 31 (14) 5325-5334
    CrossrefPubMedGoogle Scholar
  • 3 Payne Jr SH, Brushart TM. Neurotization of the rat soleus muscle: a quantitative analysis of reinnervation. J Hand Surg Am 1997; 22 (04) 640-643
    CrossrefPubMedGoogle Scholar
  • 4 Park DM, Shon SK, Kim YJ. Direct muscle neurotization in rat soleus muscle. J Reconstr Microsurg 2000; 16 (05) 379-383
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 Ma CH, Omura T, Cobos EJ. , et al. Accelerating axonal growth promotes motor recovery after peripheral nerve injury in mice. J Clin Invest 2011; 121 (11) 4332-4347
    CrossrefPubMedGoogle Scholar
  • 6 Barbour J, Yee A, Kahn LC, Mackinnon SE. Supercharged end-to-side anterior interosseous to ulnar motor nerve transfer for intrinsic musculature reinnervation. J Hand Surg Am 2012; 37 (10) 2150-2159
    CrossrefPubMedGoogle Scholar
  • 7 Midha R, Munro CA, Chan S, Nitising A, Xu QG, Gordon T. Regeneration into protected and chronically denervated peripheral nerve stumps. Neurosurgery 2005; 57 (06) 1289-1299 , discussion 1289–1299
    CrossrefPubMedGoogle Scholar
  • 8 Sulaiman OA, Gordon T. Role of chronic Schwann cell denervation in poor functional recovery after nerve injuries and experimental strategies to combat it. Neurosurgery 2009; 65 (4, Suppl): A105-A114
    CrossrefPubMedGoogle Scholar
  • 9 Sulaiman OA, Midha R, Munro CA, Matsuyama T, Al-Majed A, Gordon T. Chronic Schwann cell denervation and the presence of a sensory nerve reduce motor axonal regeneration. Exp Neurol 2002; 176 (02) 342-354
    CrossrefPubMedGoogle Scholar
  • 10 Sulaiman OA, Gordon T. Effects of short- and long-term Schwann cell denervation on peripheral nerve regeneration, myelination, and size. Glia 2000; 32 (03) 234-246
    CrossrefPubMedGoogle Scholar
  • 11 Mu L, Sobotka S, Su H. Nerve-muscle-endplate band grafting: a new technique for muscle reinnervation. Neurosurgery 2011; 69 (2, Suppl Operative): ons208-ons224 , discussion ons224
    PubMedGoogle Scholar
  • 12 Mu L, Sobotka S, Chen J, Nyirenda T. Reinnervation of denervated muscle by implantation of nerve-muscle-endplate band graft to the native motor zone of the target muscle. Brain Behav 2017; 7 (06) e00668
    CrossrefPubMedGoogle Scholar
  • 13 Sobotka S, Chen J, Nyirenda T, Mu L. Outcomes of muscle reinnervation with direct nerve implantation into the native motor zone of the target muscle. J Reconstr Microsurg 2017; 33 (02) 77-86
    PubMedGoogle Scholar
  • 14 Finkelstein DI, Dooley PC, Luff AR. Recovery of muscle after different periods of denervation and treatments. Muscle Nerve 1993; 16 (07) 769-777
    CrossrefPubMedGoogle Scholar
  • 15 Irintchev A, Draguhn A, Wernig A. Reinnervation and recovery of mouse soleus muscle after long-term denervation. Neuroscience 1990; 39 (01) 231-243
    CrossrefPubMedGoogle Scholar
  • 16 Hall ZW, Sanes JR. Synaptic structure and development: the neuromuscular junction. Cell 1993; 72 (Suppl): 99-121
    CrossrefPubMedGoogle Scholar
  • 17 Battiston B, Geuna S, Ferrero M, Tos P. Nerve repair by means of tubulization: literature review and personal clinical experience comparing biological and synthetic conduits for sensory nerve repair. Microsurgery 2005; 25 (04) 258-267
    CrossrefPubMedGoogle Scholar
  • 18 Keilhoff G, Fansa H. Successful intramuscular neurotization is dependent on the denervation period. A histomorphological study of the gracilis muscle in rats. Muscle Nerve 2005; 31 (02) 221-228
    CrossrefPubMedGoogle Scholar
  • 19 Covault J, Sanes JR. Neural cell adhesion molecule (N-CAM) accumulates in denervated and paralyzed skeletal muscles. Proc Natl Acad Sci U S A 1985; 82 (13) 4544-4548
    CrossrefPubMedGoogle Scholar
  • 20 Sanes JR, Schachner M, Covault J. Expression of several adhesive macromolecules (N-CAM, L1, J1, NILE, uvomorulin, laminin, fibronectin, and a heparan sulfate proteoglycan) in embryonic, adult, and denervated adult skeletal muscle. J Cell Biol 1986; 102 (02) 420-431
    CrossrefPubMedGoogle Scholar
  • 21 Cashman NR, Covault J, Wollman RL, Sanes JR. Neural cell adhesion molecule in normal, denervated, and myopathic human muscle. Ann Neurol 1987; 21 (05) 481-489
    CrossrefPubMedGoogle Scholar
  • 22 Müller-Felber W, Küllmer K, Fischer P. , et al. Fibre type specific expression of Leu19-antigen and N-CAM in skeletal muscle in various stages after experimental denervation. Virchows Arch A Pathol Anat Histopathol 1993; 422 (04) 277-283
    CrossrefPubMedGoogle Scholar
  • 23 Kalliainen LK, Jejurikar SS, Liang LW, Urbanchek MG, Kuzon Jr WM. A specific force deficit exists in skeletal muscle after partial denervation. Muscle Nerve 2002; 25 (01) 31-38
    CrossrefPubMedGoogle Scholar
  • 24 Mu L, Sobotka S, Chen J. , et al; Arizona Parkinson's Disease Consortium. Altered pharyngeal muscles in Parkinson disease. J Neuropathol Exp Neurol 2012; 71 (06) 520-530
    CrossrefPubMedGoogle Scholar
  • 25 Dubowitz V. Muscle Biopsy. A Practical Approach. 2nd ed. London: Bailliere Tindall; 1985
    Google Scholar
  • 26 Grumbles RM, Sesodia S, Wood PM, Thomas CK. Neurotrophic factors improve motoneuron survival and function of muscle reinnervated by embryonic neurons. J Neuropathol Exp Neurol 2009; 68 (07) 736-746
    CrossrefPubMedGoogle Scholar
  • 27 Mu L, Sobotka S, Chen J, Nyirenda T. Nerve growth factor and basic fibroblast growth factor promote reinnervation by nerve-muscle-endplate grafting. Muscle Nerve 2017; DOI: 10.1002/mus.25726.
    CrossrefPubMedGoogle Scholar
  • 28 Marques MJ, Conchello JA, Lichtman JW. From plaque to pretzel: fold formation and acetylcholine receptor loss at the developing neuromuscular junction. J Neurosci 2000; 20 (10) 3663-3675
    CrossrefPubMedGoogle Scholar
  • 29 Grossman EJ, Roy RR, Talmadge RJ, Zhong H, Edgerton VR. Effects of inactivity on myosin heavy chain composition and size of rat soleus fibers. Muscle Nerve 1998; 21 (03) 375-389
    CrossrefPubMedGoogle Scholar
  • 30 Schmalbruch H, al-Amood WS, Lewis DM. Morphology of long-term denervated rat soleus muscle and the effect of chronic electrical stimulation. J Physiol 1991; 441: 233-241
    CrossrefPubMedGoogle Scholar
  • 31 Csillik B, Nemcsók J, Chase B, Csillik AE, Knyihár-Csillik E. Infraterminal spreading and extrajunctional expression of nicotinic acetylcholine receptors in denervated rat skeletal muscle. Exp Brain Res 1999; 125 (04) 426-434
    CrossrefPubMedGoogle Scholar
  • 32 Prakash YS, Zhan WZ, Miyata H, Sieck GC. Adaptations of diaphragm neuromuscular junction following inactivity. Acta Anat (Basel) 1995; 154 (02) 147-161
    CrossrefPubMedGoogle Scholar
  • 33 Marques MJ, Mendes ZT, Minatel E, Santo Neto H. Acetylcholine receptors and nerve terminal distribution at the neuromuscular junction of long-term regenerated muscle fibers. J Neurocytol 2005; 34 (06) 387-396
    CrossrefPubMedGoogle Scholar
  • 34 Frank E, Gautvik K, Sommerschild H. Cholinergic receptors at denervated mammalian motor end-plates. Acta Physiol Scand 1975; 95 (01) 66-76
    CrossrefPubMedGoogle Scholar
  • 35 Szabo M, Salpeter EE, Randall W, Salpeter MM. Transients in acetylcholine receptor site density and degradation during reinnervation of mouse sternomastoid muscle. J Neurochem 2003; 84 (01) 180-188
    PubMedGoogle Scholar
  • 36 Fumagalli G, Balbi S, Cangiano A, Lømo T. Regulation of turnover and number of acetylcholine receptors at neuromuscular junctions. Neuron 1990; 4 (04) 563-569
    CrossrefPubMedGoogle Scholar
  • 37 Kumai Y, Ito T, Matsukawa A, Yumoto E. Effects of denervation on neuromuscular junctions in the thyroarytenoid muscle. Laryngoscope 2005; 115 (10) 1869-1872
    CrossrefPubMedGoogle Scholar
  • 38 Birks R, Katz B, Miledi R. Physiological and structural changes at the amphibian myoneural junction, in the course of nerve degeneration. J Physiol 1960; 150: 145-168
    CrossrefPubMedGoogle Scholar
  • 39 Frank E, Gautvik K, Sommerschild H. Persistence of junctional acetylcholine receptors following denervation. Cold Spring Harb Symp Quant Biol 1976; 40: 275-281
    CrossrefPubMedGoogle Scholar
  • 40 Sanes JR, Marshall LM, McMahan UJ. Reinnervation of skeletal muscle: restoration of the normal synaptic pattern. In: Jewett DL, McCarroll HR. , eds. Nerve Repair: Its Clinical and Experimental Basis. St. Louis (MO): Mosby; 1980
    Google Scholar
  • 41 Steinbach JH. Neuromuscular junctions and alpha-bungarotoxin-binding sites in denervated and contralateral cat skeletal muscles. J Physiol 1981; 313: 513-528
    CrossrefPubMedGoogle Scholar
  • 42 Sanes JR, Lichtman JW. Development of the vertebrate neuromuscular junction. Annu Rev Neurosci 1999; 22: 389-442
    CrossrefPubMedGoogle Scholar
  • 43 Bassel-Duby R, Olson EN. Signaling pathways in skeletal muscle remodeling. Annu Rev Biochem 2006; 75: 19-37
    CrossrefPubMedGoogle Scholar
  • 44 Wu H, Xiong WC, Mei L. To build a synapse: signaling pathways in neuromuscular junction assembly. Development 2010; 137 (07) 1017-1033
    CrossrefPubMedGoogle Scholar
  • 45 Iwayama T. Relation of regenerating nerve terminals to original endplates. Nature 1969; 224 (5214): 81-82
    CrossrefPubMedGoogle Scholar
  • 46 Bennett MR, McLachlan EM, Taylor RS. The formation of synapses in reinnervated mammalian striated muscle. J Physiol 1973; 233 (03) 481-500
    CrossrefPubMedGoogle Scholar
  • 47 Sanes JR, Marshall LM, McMahan UJ. Reinnervation of muscle fiber basal lamina after removal of myofibers. Differentiation of regenerating axons at original synaptic sites. J Cell Biol 1978; 78 (01) 176-198
    CrossrefPubMedGoogle Scholar
  • 48 Gorio A, Carmignoto G, Finesso M, Polato P, Nunzi MG. Muscle reinnervation--II. Sprouting, synapse formation and repression. Neuroscience 1983; 8 (03) 403-416
    CrossrefPubMedGoogle Scholar
  • 49 Covault J, Cunningham JM, Sanes JR. Neurite outgrowth on cryostat sections of innervated and denervated skeletal muscle. J Cell Biol 1987; 105 (6 Pt 1): 2479-2488
    CrossrefPubMedGoogle Scholar
  • 50 Guth L, Zalewski AA. Disposition of cholinesterase following implantation of nerve into innervated and denervated muscle. Exp Neurol 1963; 7: 316-326
    CrossrefPubMedGoogle Scholar
  • 51 Sakellarides HT, Sorbie C, James L. Reinnervation of denervated muscles by nerve transplantation. Clin Orthop Relat Res 1972; 83: 195-201
    PubMedGoogle Scholar
  • 52 McMahan UJ, Wallace BG. Molecules in basal lamina that direct the formation of synaptic specializations at neuromuscular junctions. Dev Neurosci 1989; 11 (4-5): 227-247
    CrossrefPubMedGoogle Scholar
  • 53 Schwarting S, Schröder M, Stennert E, Goebel HH. Morphology of denervated human facial muscles. ORL J Otorhinolaryngol Relat Spec 1984; 46 (05) 248-256
    CrossrefPubMedGoogle Scholar
  • 54 Gutmann E, Young JZ. The re-innervation of muscle after various periods of atrophy. J Anat 1944; 78 (Pt 1-2): 15-43
    PubMedGoogle Scholar
  • 55 Hynes NM, Bain JR, Thoma A, Veltri K, Maguire JA. Preservation of denervated muscle by sensory protection in rats. J Reconstr Microsurg 1997; 13 (05) 337-343
    Article in Thieme ConnectPubMedGoogle Scholar
  • 56 Bain JR, Veltri KL, Chamberlain D, Fahnestock M. Improved functional recovery of denervated skeletal muscle after temporary sensory nerve innervation. Neuroscience 2001; 103 (02) 503-510
    CrossrefPubMedGoogle Scholar
  • 57 Beck-Broichsitter BE, Becker ST, Lamia A, Fregnan F, Geuna S, Sinis N. Sensoric protection after median nerve injury: babysitter-procedure prevents muscular atrophy and improves neuronal recovery. Biomed Res Int 2014; 2014: 724197 , 7 pages
    PubMedGoogle Scholar
  • 58 Wang H, Gu Y, Xu J, Shen L, Li J. Comparative study of different surgical procedures using sensory nerves or neurons for delaying atrophy of denervated skeletal muscle. J Hand Surg Am 2001; 26 (02) 326-331
    PubMedGoogle Scholar
  • 59 Noordin S, Ahmed M, Rehman R, Ahmad T, Hashmi P. Neuronal regeneration in denervated muscle following sensory and muscular neurotization. Acta Orthop 2008; 79 (01) 126-133
    CrossrefPubMedGoogle Scholar
  • 60 Veltri K, Kwiecien JM, Minet W, Fahnestock M, Bain JR. Contribution of the distal nerve sheath to nerve and muscle preservation following denervation and sensory protection. J Reconstr Microsurg 2005; 21 (01) 57-70 , discussion 71–74
    Article in Thieme ConnectPubMedGoogle Scholar
  • 61 Hosseinian MA, Shirian S, Loron AG, Ebrahimy AA. , Hayatolah1 GH. Distal sensory to distal motor nerve anastomosis can protect lower extremity muscle atrophy in a murine model. Eur J Plast Surg 2017; DOI: 10.1007/s00238-017-1313-z.
    CrossrefPubMedGoogle Scholar
  • 62 Papakonstantinou KC, Kamin E, Terzis JK. Muscle preservation by prolonged sensory protection. J Reconstr Microsurg 2002; 18 (03) 173-182 , discussion 183–184
    Article in Thieme ConnectPubMedGoogle Scholar
  • 63 Bain JR, Hason Y, Veltri K, Fahnestock M, Quartly C. Clinical application of sensory protection of denervated muscle. J Neurosurg 2008; 109 (05) 955-961
    CrossrefPubMedGoogle Scholar
  • 64 Nghiem BT, Sando IC, Hu Y, Urbancheck MG, Cederna PS. Sensory protection to enhance functional recovery following proximal nerve injuries: current trends. Plast Aesthet Res 2015; 2: 202-207
    CrossrefPubMedGoogle Scholar
  • 65 Bader D. Reinnervation of motor endplate-containing and motor endplate-less muscle grafts. Dev Biol 1980; 77 (02) 315-327
    CrossrefPubMedGoogle Scholar
  • 66 Grosheva M, Nohroudi K, Schwarz A. , et al. Comparison of trophic factors' expression between paralyzed and recovering muscles after facial nerve injury. A quantitative analysis in time course. Exp Neurol 2016; 279: 137-148
    CrossrefPubMedGoogle Scholar
  • 67 Fox MA, Sanes JR, Borza DB. , et al. Distinct target-derived signals organize formation, maturation, and maintenance of motor nerve terminals. Cell 2007; 129 (01) 179-193
    CrossrefPubMedGoogle Scholar
  • 68 Sakuma K, Yamaguchi A. The recent understanding of the neurotrophin's role in skeletal muscle adaptation. J Biomed Biotechnol 2011; 2011: 201696
    PubMedGoogle Scholar
  • 69 Zahavi EE, Ionescu A, Gluska S, Gradus T, Ben-Yaakov K, Perlson E. A compartmentalized microfluidic neuromuscular co-culture system reveals spatial aspects of GDNF functions. J Cell Sci 2015; 128 (06) 1241-1252
    CrossrefPubMedGoogle Scholar
  • 70 Wang J, Sun J, Tang Y. , et al. Basic fibroblast growth factor attenuates the degeneration of injured spinal cord motor endplates. Neural Regen Res 2013; 8 (24) 2213-2224
    PubMedGoogle Scholar
  • 71 Yang LX, Nelson PG. Glia cell line-derived neurotrophic factor regulates the distribution of acetylcholine receptors in mouse primary skeletal muscle cells. Neuroscience 2004; 128 (03) 497-509
    CrossrefPubMedGoogle Scholar
  • 72 Suzuki M, McHugh J, Tork C. , et al. Direct muscle delivery of GDNF with human mesenchymal stem cells improves motor neuron survival and function in a rat model of familial ALS. Mol Ther 2008; 16 (12) 2002-2010
    CrossrefPubMedGoogle Scholar
  • 73 Sobotka S, Chen J, Nyirenda T, Mu L. Intraoperative 1-hour electrical nerve stimulation enhances outcomes of nerve-muscle-endplate band grafting technique for muscle reinnervation. J Reconstr Microsurg 2017; 33 (08) 533-543
    Article in Thieme ConnectPubMedGoogle Scholar