Cloning of cDNA encoding a regeneration-associated muscle protease whose expression is attenuated in cell lines derived from Duchenne muscular dystrophy patients
Cloning of cDNA encoding a regeneration-associated muscle protease whose expression is attenuated in cell lines derived from Duchenne muscular dystrophy patients
复制标题
DOI：
10.1016/s0002-9440(10)63735-2
复制
发表时间：
2004-05-01
影响因子：
6
通讯作者：
Hara, T
中科院分区：
文献类型：
--
作者：
Nakayama, Y;Nara, N;Hara, T
在肌营养不良蛋白突变型mdx小鼠（杜氏肌营养不良（DMD）的动物模型）中，受损的骨骼肌被有效地再生，因此动物茁壮成长。DMD患者和mdx小鼠之间的表型差异表明存在调节mdx小鼠肌肉萎缩的因素。为了确定这些因素，我们使用cDNA微阵列与新建立的骨骼肌细胞系mdx和正常小鼠的mdx突变的影响，寻找mRNA。我们发现，在mdx肌肉细胞系中，12个基因，包括L-精氨酸：甘氨酸脒基转移酶和胸腺素β 4，上调，而7个基因，包括硒蛋白P和一种新的再生相关肌肉蛋白酶（RAMP），下调。北方印迹分析和原位杂交结果显示，RAMP mRNA主要表达于正常小鼠骨骼肌和脑组织，在小鼠骨骼肌损伤后再生区表达增强。与正常肌肉细胞系相比，来自6名DMD患者活检的单个肌肉细胞系中的RAMP表达要低得多。这些结果表明，RAMP可能在骨骼肌再生中发挥作用，其下调可能参与人类DMD的进展。
in the dystrophin-mutant mdx mouse, an animal model for Duchenne muscular dystrophy (DMD), damaged skeletal muscles are efficiently regenerated and thus the animals thrive. The phenotypic differences between DMD patients and the mdx mice suggest the existence of factors that modulate the muscle wasting in die mdx mice. To identify these factors, we searched for mRNAs affected by the mdx mutation by using cDNA microarrays with newly established skeletal muscle cell lines from mdx and normal mice. We found that in the mdx muscle cell line, 12 genes, including L-arginine:glycine amidinotransferase and thymosin beta4, are up-regulated, whereas 7 genes, including selenoprotein P and a novel regeneration-associated muscle protease (RAMP), are down-regulated. Northern blot analysis and in situ hybridization revealed that RAMP mRNA is predominantly expressed in normal skeletal muscle and brain, and its production is enhanced in the regenerating area of injured skeletal muscle in mice. RAMP expression was much lower in individual muscle cell lines derived from biopsies of six DMD patients compared to a normal muscle cell line. These results suggest that RAMP may play a role in the regeneration of skeletal muscle and that its down-regulation could be involved in the progression of DMD in humans.