Department of Neurology, Third Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
Department of Neurology, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
Neural Regen Res. 2014 Feb 15;9(4):413-9. doi: 10.4103/1673-5374.128248.
We previously found that the K141N mutation in heat shock protein B8 (HSPB8) was responsible for Charcot-Marie-Tooth disease type 2L in a large Chinese family. The objective of the present study was to generate a transgenic mouse model bearing the K141N mutation in the human HSPB8 gene, and to determine whether this (K141N)HSPB8 transgenic mouse model would manifest the clinical phenotype of Charcot-Marie-Tooth disease type 2L, and consequently be suitable for use in studies of disease pathogenesis. Transgenic mice overexpressing (K141N)HSPB8 were generated using K141N mutant HSPB8 cDNA cloned into a pCAGGS plasmid driven by a human cytomegalovirus expression system. PCR and western blot analysis confirmed integration of the (K141N)HSPB8 gene and widespread expression in tissues of the transgenic mice. The (K141N)HSPB8 transgenic mice exhibited decreased muscle strength in the hind limbs and impaired motor coordination, but no obvious sensory disturbance at 6 months of age by behavioral assessment. Electrophysiological analysis showed that the compound motor action potential amplitude in the sciatic nerve was significantly decreased, but motor nerve conduction velocity remained normal at 6 months of age. Pathological analysis of the sciatic nerve showed reduced myelinated fiber density, notable axonal edema and vacuolar degeneration in (K141N)HSPB8 transgenic mice, suggesting axonal involvement in the peripheral nerve damage in these animals. These findings indicate that the (K141N)HSPB8 transgenic mouse successfully models Charcot-Marie-Tooth disease type 2L and can be used to study the pathogenesis of the disease.
我们之前发现热休克蛋白 B8 (HSPB8) 的 K141N 突变是一个中国大家族中 2L 型腓骨肌萎缩症的致病原因。本研究的目的是构建携带人类 HSPB8 基因 K141N 突变的转基因小鼠模型,确定该 (K141N)HSPB8 转基因小鼠模型是否会表现出 2L 型腓骨肌萎缩症的临床表型,进而适合用于疾病发病机制的研究。使用克隆到 pCAGGS 质粒的 HSPB8 K141N 突变 cDNA,通过人巨细胞病毒表达系统驱动,构建了过表达 (K141N)HSPB8 的转基因小鼠。PCR 和 Western blot 分析证实了 (K141N)HSPB8 基因的整合及其在转基因小鼠组织中的广泛表达。在 6 月龄时,(K141N)HSPB8 转基因小鼠表现出后肢肌肉力量减弱和运动协调能力受损,但行为评估未显示出明显的感觉障碍。电生理学分析显示,坐骨神经的复合运动动作电位幅度明显降低,但运动神经传导速度在 6 月龄时仍正常。坐骨神经的病理学分析显示,(K141N)HSPB8 转基因小鼠的有髓纤维密度降低,轴突水肿和空泡变性明显,提示这些动物的周围神经损伤存在轴突受累。这些发现表明,(K141N)HSPB8 转基因小鼠成功模拟了 2L 型腓骨肌萎缩症,可用于研究该疾病的发病机制。