Increased expression of rapsyn in muscles prevents acetylcholine receptor loss in experimental autoimmune myasthenia gravis

• Published in: Brain (London, England : 1878) England : 1878), 2005-10, Vol.128 (10), p.2327-2337
• Main Authors: Losen, Mario, Stassen, Maurice H. W., Martínez-Martínez, Pilar, Machiels, Barbie M., Duimel, Hans, Frederik, Peter, Veldman, Henk, Wokke, John H. J., Spaans, Frank, Vincent, Angela, De Baets, Marc H.
• Format: Article
• Language: English
• Subjects: AChR = acetylcholine receptor; Action Potentials - physiology; Acute Disease; Animals; Biological and medical sciences; BN rat = Brown Norway rat; CMAP = compound muscle action potential; Diseases of striated muscles. Neuromuscular diseases; EAMG = experimental autoimmune myasthenia gravis; Electromyography - methods; electropermeabilization; EMG = electromyography; experimental autoimmune myasthenia gravis (EAMG); Female; Fluorescent Antibody Technique - methods; gene therapy; Immunohistochemistry - methods; in vivo electroporation; Injuries of the nervous system and the skull. Diseases due to physical agents; mAb = monoclonal antibody; MAC = membrane attack complex; Medical sciences; Microscopy, Confocal - methods; Microscopy, Electron - methods; Muscle Proteins - analysis; Muscle Proteins - genetics; Muscle, Skeletal - pathology; Muscle, Skeletal - physiopathology; MuSK = muscle-specific kinase; Myasthenia Gravis, Autoimmune, Experimental - genetics; Myasthenia Gravis, Autoimmune, Experimental - pathology; Myasthenia Gravis, Autoimmune, Experimental - physiopathology; Neurology; Neuromuscular Junction - genetics; Neuromuscular Junction - pathology; Neuromuscular Junction - physiopathology; NMJ = neuromuscular junction; Radioimmunoassay - methods; rapsyn; Rats; Rats, Inbred Lew; Receptors, Cholinergic - genetics; RIA = radioimmunoassay; Synaptic Transmission - genetics; Synaptic Transmission - physiology; Traumas. Diseases due to physical agents; Up-Regulation - genetics; VAChT = vesicular acetylcholine transporter; α-BT = α-bungarotoxin
• ISSN: 0006-8950; 1460-2156
• DOI: 10.1093/brain/awh612

Myasthenia gravis is usually caused by autoantibodies to the acetylcholine receptor (AChR). The AChR is clustered and anchored in the postsynaptic membrane of the neuromuscular junction (NMJ) by a cytoplasmic protein called rapsyn. We previously showed that resistance to experimental autoimmune myasthenia gravis (EAMG) in aged rats correlates with increased rapsyn concentration at the NMJ. It is possible, therefore, that endogenous rapsyn expression may be an important determinant of AChR loss and neuromuscular transmission failure in the human disease, and that upregulation of rapsyn expression could be used therapeutically. To examine first a potential therapeutic application of rapsyn upregulation, we induced acute EAMG in young rats by passive transfer of AChR antibody, mAb 35, and used in vivo electroporation to over-express rapsyn unilaterally in one tibialis anterior. We looked at the compound muscle action potentials (CMAPs) in the tibialis anterior, at rapsyn and AChR expression by quantitative radioimmunoassay and immunofluorescence, and at the morphology of the NMJs, comparing the electroporated and untreated muscles, as well as the control and EAMG rats. In control rats, transfected muscle fibres had extrasynaptic rapsyn aggregates, as well as slightly increased rapsyn and AChR concentrations at the NMJ. In EAMG rats, despite deposits of the membrane attack complex, the rapsyn-overexpressing muscles showed no decrement in the CMAPs, no loss of AChR, and the majority had normal postsynaptic folds, whereas endplates of untreated muscles showed typical AChR loss and morphological damage. These data suggest not only that increasing rapsyn expression could be a potential treatment for selected muscles of myasthenia gravis patients, but also lend support to the hypothesis that individual differences in innate rapsyn expression could be a factor in determining disease severity.