Proteasome inhibitors suppress formation of polyglutamine‐induced nuclear inclusions in cultured postmitotic neurons
Proteasome inhibitors suppress formation of polyglutamine‐induced nuclear inclusions in cultured postmitotic neurons
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蛋白酶体抑制剂抑制培养的有丝分裂后神经元中聚谷氨酰胺诱导的核包涵体的形成
作者：
Woo;C. Horbinski;W. Sigurdson;D. Higgins
至少有九种神经退行性疾病是由不同基因中多聚谷氨酰胺重复序列的扩张引起的。这种扩张导致在不同类型的细胞内形成核包涵体(NI)。在这项研究中，我们利用出生前大鼠颈上神经节的交感神经元原代培养建立了一种多谷氨酰胺病的模型。将编码127个谷氨酰胺重复序列的质粒导入∼，可使70%的交感神经元在6 天内发生NI。此外，它还会导致躯体萎缩，抑制树突生长。NIS含有泛素化蛋白，并隔离分子伴侣热休克蛋白70(Hsp70)。我们发现，两种特异的蛋白酶体抑制剂lactacystin和CEP1612可以抑制多聚谷氨酰胺诱导的NI的形成。此外，lactacystin处理诱导了先前存在的NI的清除。Western blotting和免疫细胞化学显示，lactacystin和CEP1612强烈诱导Hsp70的表达，而非特异性的蛋白酶体抑制剂N-乙酰-亮氨酸-去亮氨酸不能诱导Hsp70的表达。将127个谷氨酸与编码野生型Hsp70基因的质粒共表达后，含有NI的神经元百分比显著降低。此外，用编码突变型Hsp70的质粒转染可阻断lactacystin的作用。这些发现进一步表明Hsp70是一种神经保护分子，并表明某些蛋白酶体抑制剂在治疗多谷氨酰胺疾病方面具有潜在的实用价值。
At least nine neurodegenerative disorders are caused by expansion of polyglutamine repeats in various genes. This expansion induces the formation of nuclear inclusions (NI) within various cell types. In this study, we developed a model for polyglutamine diseases using primary cultures of sympathetic neurons from the superior cervical ganglia of prenatal rat pups. Transfection with a plasmid encoding 127 glutamine repeats causes NI to develop in ∼70% of the sympathetic neurons within 6 days. In addition, it causes somatic atrophy and inhibits dendritic growth. The NIs contain ubiquitinated proteins and sequester the molecular chaperone heat shock protein 70 (Hsp70). We found that two specific proteasome inhibitors, lactacystin and CEP1612, suppress thezformation of polyglutamine‐induced NI. In addition, lactacystin treatment induced the removal of preexisting NI. Western blotting and immunocytochemistry revealed that lactacystin and CEP1612 strongly induce the expression of Hsp70, whereas less specific proteasome inhibitor such as N‐acetyl‐Leu‐Leu‐Norleucinal does not. Coexpression of 127 glutamines with a plasmid encoding wild‐type Hsp70 gene resulted in a marked reduction of the percentage of neurons containing NI. In addition, transfection with plasmids encoding mutant Hsp70 blocked the effects of lactacystin. These findings further implicate Hsp70 as a neuroprotective molecule and they suggest the potential utility of certain proteasome inhibitors in the treatment of polyglutamine diseases.
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大鼠交感神经元在组织培养中形成树突的能力随年龄变化。
DOI：
10.1016/0165-3806(89)90140-5
发表时间：
1989
期刊：
Brain research. Developmental brain research
影响因子：
--
作者：
Bruckenstein,D;Johnson,MI;Higgins,D
通讯作者：
Higgins,D
影响因子：
3.5
作者：
Chan, HYE;Warrick, JM;Bonini, NM
通讯作者：
Bonini, NM
影响因子：
3.5
作者：
Marsh, JL;Walker, H;Thompson, LM
通讯作者：
Thompson, LM
影响因子：
56.9
作者：
DiFiglia, M;Sapp, E;Aronin, N
通讯作者：
Aronin, N
影响因子：
3.5
作者：
Cummings, CJ;Sun, YL;Zoghbi, HY
通讯作者：
Zoghbi, HY