DOPAL is transmissible to and oligomerizes alpha-synuclein in human glial cells.
DOPAL is transmissible to and oligomerizes alpha-synuclein in human glial cells.
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DOI：
10.1016/j.autneu.2015.12.008
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发表时间：
2016-01
期刊：
影响因子：
--
通讯作者：
Goldstein DS
中科院分区：
文献类型：
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作者：
Jinsmaa Y;Sullivan P;Sharabi Y;Goldstein DS
含有α -突触核蛋白（AS）的胶质细胞质内含物（GCIs）是多系统萎缩（MSA）的神经病理学标志。寡聚的AS被认为是该蛋白的致病性形式。胶质细胞通常表达少量AS，但它们可以从细胞外液中摄取AS。3,4-二羟基苯乙醛（DOPAL）是多巴胺（DA）神经元内代谢的专性中间体，能有效地使AS寡聚。在这项研究中，我们测试了DOPAL是否被人胶质细胞吸收并增加细胞内AS的寡聚化。将DOPAL（与PC12细胞共孵育的外源性或内源性）和AS（天然或A53T突变形式）添加到胶质细胞（胶质母细胞瘤或MO3.13少突胶质细胞）的孵育培养基中。在长达180分钟的孵育下，测量DOPAL及其细胞内代谢物3,4-二羟基苯乙酸（DOPAC）的胶质细胞含量。免疫印迹法定量测定胶质细胞AS低聚物。胶质母细胞瘤和MO3.13细胞均不含内源性儿茶酚胺或AS。与产生da的PC12细胞共孵育后，DOPAL和DOPAC含量随时间增加。同样，在培养基中加入DOPAL后，胶质细胞DOPAL和DOPAC含量迅速增加。添加天然或A53T-AS后，细胞内AS也增加。与DOPAL和AS共同孵育的胶质细胞增强了细胞内天然和A53T-AS的寡聚化。DOPAL可传递到胶质细胞，并增强AS的细胞内寡聚化。DOPAL与AS的相互作用可能有助于解释MSA中cgi的形成。
Glial cytoplasmic inclusions (GCIs) containing alpha-synuclein (AS) are a neuropathologic hallmark of multiple system atrophy (MSA). Oligomerized AS is thought to be the pathogenic form of the protein. Glial cells normally express little AS, but they can take up AS from the extracellular fluid. 3,4-dihydroxyphenylacetaldehyde (DOPAL), an obligate intermediate in the intra-neuronal metabolism of dopamine (DA), potently oligomerizes AS. In this study we tested whether DOPAL is taken up by human glial cells and augments intracellular oligomerization of AS. DOPAL (exogenous or endogenous from co-incubation with PC12 cells) and AS (native or A53T mutant form) were added to the incubation medium of glial cells (glioblastoma or MO3.13 oligodendrocytes). Glial cellular contents of DOPAL and its intracellular metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) were measured at up to 180 minutes of incubation. Glial cellular AS oligomers were quantified by Western blotting. Neither glioblastoma nor MO3.13 cells contained endogenous catecholamines or AS. Co-incubation of the cells with DA-producing PC12 cells produced time-related increases in DOPAL and DOPAC contents. Similarly, glial cellular DOPAL and DOPAC contents increased rapidly after addition of DOPAL to the medium. After addition of native or A53T-AS, intracellular AS also increased. Incubation of glial cells with both DOPAL and AS enhanced the intracellular oligomerization of native and A53T-AS. DOPAL is transmissible to glial cells and enhances intracellular oligomerization of AS. An interaction of DOPAL with AS might help explain the formation of CGIs in MSA.