Hegedus J, Putman C T, Tyreman N, Gordon T
Centre for Neuroscience, 525 Heritage Medical Research Centre, Faculty of Medicine, University of Alberta, Edmonton, AB, Canada.
J Physiol. 2008 Jul 15;586(14):3337-51. doi: 10.1113/jphysiol.2007.149286. Epub 2008 May 8.
The present study investigated motor unit (MU) loss in a murine model of familial amyotrophic lateral sclerosis (ALS). The fast-twitch tibialis anterior (TA) and medial gastrocnemius (MG) muscles of transgenic SOD1(G93A) and SOD1(WT) mice were studied during the presymptomatic phase of disease progression at 60 days of age. Whole muscle maximum isometric twitch and tetanic forces were 80% lower (P < 0.01) in the TA muscles of SOD1(G93A) compared to SOD1(WT) mice. Enumeration of total MU numbers within TA muscles showed a 60% reduction (P < 0.01) within SOD1(G93A) mice (38 +/- 7) compared with SOD1(WT) controls (95 +/- 12); this was attributed to a lower proportion of the most forceful fast-fatigable (FF) MU in SOD1(G93A) mice, as seen by a significant (P < 0.01) leftward shift in the cumulative frequency histogram of single MU forces. Similar patterns of MU loss and corresponding decreases in isometric twitch force were observed in the MG. Immunocytochemical analyses of the entire cross-sectional area (CSA) of serial sections of TA muscles stained with anti-neural cell adhesion molecule (NCAM) and various monoclonal antibodies for myosin heavy chain (MHC) isoforms showed respective 65% (P < 0.01) and 28% (P < 0.05) decreases in the number of innervated IIB and IID/X muscle fibres in SOD1(G93A), which paralleled the 60% decrease (P < 0.01) in the force generating capacity of individual fibres. The loss of fast MUs was partially compensated by activity-dependent fast-to-slower fibre type transitions, as determined by increases (P < 0.04) in the CSA and proportion of IIA fibres (from 4% to 14%) and IID/X fibres (from 31% to 39%), and decreases (P < 0.001) in the CSA and proportion of type IIB fibres (from 65% to 44%). We conclude that preferential loss of IIB fibres is incomplete at 60 days of age, and is consistent with a selective albeit gradual loss of FF MUs that is not fully compensated by sprouting of the remaining motoneurons that innervate type IIA or IID/X muscle fibres. Our findings indicate that disease progression in fast-twitch muscles of SOD1(G93A) mice involves parallel processes: (1) gradual selective motor axon die-back of the FF motor units that contain large type IIB muscle fibres, and of fatigue-intermediate motor units that innervate type IID/X muscle fibres, and (2) activity-dependent conversion of motor units to those innervated by smaller motor axons innervating type IIA fatigue-resistant muscle fibres.
本研究在家族性肌萎缩侧索硬化症(ALS)小鼠模型中调查了运动单位(MU)的损失情况。在60日龄疾病进展的症状前期,对转基因SOD1(G93A)小鼠和SOD1(野生型)小鼠的快肌胫骨前肌(TA)和腓肠肌内侧头(MG)进行了研究。与SOD1(野生型)小鼠相比,SOD1(G93A)小鼠TA肌的全肌最大等长收缩抽搐力和强直收缩力降低了80%(P < 0.01)。对TA肌内总运动单位数量的计数显示,与SOD1(野生型)对照(95 ± 12)相比,SOD1(G93A)小鼠的运动单位数量减少了60%(P < 0.01)(38 ± 7);这归因于SOD1(G93A)小鼠中最有力的快速易疲劳(FF)运动单位比例较低,这在单个运动单位力的累积频率直方图中表现为显著(P < 0.01)向左偏移。在MG中也观察到了类似的运动单位损失模式以及等长收缩抽搐力的相应降低。用抗神经细胞黏附分子(NCAM)和各种肌球蛋白重链(MHC)亚型单克隆抗体对TA肌连续切片的整个横截面积(CSA)进行免疫细胞化学分析显示,SOD1(G93A)小鼠中受神经支配的IIB型和IID/X型肌纤维数量分别减少了65%(P < 0.01)和28%(P < 0.05),这与单个纤维产生力的能力降低60%(P < 0.01)相平行。快速运动单位的损失部分通过活动依赖的快速纤维类型向慢速纤维类型转变得到补偿,这表现为IIA型纤维(从4%增加到14%)和IID/X型纤维(从31%增加到39%)的CSA和比例增加(P < 0.04),以及IIB型纤维的CSA和比例减少(P < 0.001)(从65%减少到44%)。我们得出结论,在60日龄时,IIB型纤维的优先损失并不完全,这与FF运动单位的选择性(尽管是逐渐的)损失一致,而这种损失并没有通过支配IIA型或IID/X型肌纤维的其余运动神经元的芽生得到完全补偿。我们的研究结果表明,SOD1(G93A)小鼠快肌中的疾病进展涉及平行过程:(1)包含大型IIB型肌纤维的FF运动单位以及支配IID/X型肌纤维的疲劳中间型运动单位的逐渐选择性运动轴突回缩,以及(2)运动单位向由支配IIA型抗疲劳肌纤维的较小运动轴突支配的运动单位的活动依赖转变。