Patel Vishwendra, Oh Anne, Voit Antanina, Sultatos Lester G, Babu Gopal J, Wilson Brenda A, Ho Mengfei, McArdle Joseph J
Department of Pharmacology and Physiology, New Jersey Medical School-Rutgers University, Newark, New Jersey, United States of America.
Department Cell Biology and Molecular Medicine, New Jersey Medical School-Rutgers University, Newark, New Jersey, United States of America.
PLoS One. 2014 Dec 1;9(12):e110571. doi: 10.1371/journal.pone.0110571. eCollection 2014.
Recent studies demonstrate reduced motor-nerve function during autoimmune muscle-specific tyrosine kinase (MuSK) myasthenia gravis (MG). To further understand the basis of motor-nerve dysfunction during MuSK-MG, we immunized female C57/B6 mice with purified rat MuSK ectodomain. Nerve-muscle preparations were dissected and neuromuscular junctions (NMJs) studied electrophysiologically, morphologically, and biochemically. While all mice produced antibodies to MuSK, only 40% developed respiratory muscle weakness. In vitro study of respiratory nerve-muscle preparations isolated from these affected mice revealed that 78% of NMJs produced endplate currents (EPCs) with significantly reduced quantal content, although potentiation and depression at 50 Hz remained qualitatively normal. EPC and mEPC amplitude variability indicated significantly reduced number of vesicle-release sites (active zones) and reduced probability of vesicle release. The readily releasable vesicle pool size and the frequency of large amplitude mEPCs also declined. The remaining NMJs had intermittent (4%) or complete (18%) failure of neurotransmitter release in response to 50 Hz nerve stimulation, presumably due to blocked action potential entry into the nerve terminal, which may arise from nerve terminal swelling and thinning. Since MuSK-MG-affected muscles do not express the AChR γ subunit, the observed prolongation of EPC decay time was not due to inactivity-induced expression of embryonic acetylcholine receptor, but rather to reduced catalytic activity of acetylcholinesterase. Muscle protein levels of MuSK did not change. These findings provide novel insight into the pathophysiology of autoimmune MuSK-MG.
近期研究表明,在自身免疫性肌肉特异性酪氨酸激酶(MuSK)重症肌无力(MG)期间运动神经功能会降低。为了进一步了解MuSK-MG期间运动神经功能障碍的基础,我们用纯化的大鼠MuSK胞外域免疫雌性C57/B6小鼠。解剖神经-肌肉标本,并从电生理、形态学和生物化学方面研究神经肌肉接头(NMJ)。虽然所有小鼠都产生了针对MuSK的抗体,但只有40%出现呼吸肌无力。对从这些受影响小鼠分离出的呼吸神经-肌肉标本进行的体外研究显示,78%的NMJ产生的终板电流(EPC)的量子含量显著降低,尽管在50Hz时的增强和抑制在性质上仍正常。EPC和微小终板电流(mEPC)幅度变异性表明囊泡释放位点(活性区)数量显著减少,囊泡释放概率降低。易于释放的囊泡池大小和大幅度mEPC的频率也下降。其余的NMJ在50Hz神经刺激时出现间歇性(4%)或完全(18%)的神经递质释放失败,这可能是由于动作电位进入神经末梢受阻,这可能是由神经末梢肿胀和变细引起的。由于MuSK-MG影响的肌肉不表达乙酰胆碱受体(AChR)γ亚基,观察到的EPC衰减时间延长不是由于胚胎型乙酰胆碱受体的无活性诱导表达,而是由于乙酰胆碱酯酶的催化活性降低。MuSK的肌肉蛋白水平没有变化。这些发现为自身免疫性MuSK-MG的病理生理学提供了新的见解。