In situ hybridization for detection of nocardial 16S rRNA: reactivity within intracellular inclusions in experimentally infected cynomolgus monkeys - and in Lewy body-containing human brain specimens
In situ hybridization for detection of nocardial 16S rRNA: reactivity within intracellular inclusions in experimentally infected cynomolgus monkeys - and in Lewy body-containing human brain specimens
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DOI：
10.1016/s0014-4886(03)00337-6
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发表时间：
2003-12-01
影响因子：
5.3
通讯作者：
LeWitt, PA
中科院分区：
文献类型：
--
作者：
Chapman, G;Beaman, BL;LeWitt, PA
我们以前的研究发现，实验感染的BALB/c小鼠与革兰氏阳性菌星状诺卡氏菌诱导帕金森氏综合征型左旋多巴反应性运动异常，黑质纹状体多巴胺能神经元的损失，纹状体多巴胺耗竭，和神经元内包涵体在黑质（SN）与路易体的外观相似。在本研究中，开发了一种原位杂交技术，使用诺卡氏菌特异性探针（1377）检测诺卡氏菌16 S核糖体RNA（rRNA）。在实验性感染N.小行星杂交反应检测内诺卡氏菌感染后48小时内的结构和细胞内包涵体（免疫反应的α-突触核蛋白和泛素）在两个3.5个月感染的猴子之一。然后以盲法将原位杂交程序应用于24个具有Lewy小体的人SN标本和11个不具有Lewy小体的人SN标本（包括5个正常对照）。杂交反应中检测到9个路易体含有标本，没有其他的。反应性仅限于路易体外观的包涵体，但在一份标本中也观察到诺卡氏菌样结构的细胞内反应性。这些结果表明，诺卡氏菌和神经退行性疾病，其中路易体的存在之间可能存在关联。(C)2003年爱思唯尔公司All rights reserved.
Our previous studies found that experimental infection of BALB/c mice with the Gram-positive bacterium Nocardia asteroides induced a parkinsonian-type syndrome with levodopa-responsive movement abnormalities, loss of nigrostriatal dopaminergic neurons, depletion of striatal dopamine, and intraneuronal inclusions in the substantia nigra (SN) with an appearance similar to Lewy bodies. In the present study, an in situ hybridization technique was developed to detect nocardial 16S ribosomal RNA (rRNA), using a Nocardia-specific probe (1377). Cerebral cortical specimens from cynomolgus monkeys were examined for the presence of nocardial RNA 48 h, 3.5 months, and I year after experimental infection with N. asteroides. Hybridization reactions were detected within Nocardia-like structures 48 h after infection and within intracellular inclusion bodies (immunoreactive for a-synuclein and ubiquitin) in one of two 3.5-month-infected monkeys. The in situ hybridization procedure was then applied in a blinded fashion to 24 human SN specimens with Lewy bodies and 11 human SN specimens without Lewy bodies (including five normal controls). Hybridization reactions were detected in nine Lewy body-containing specimens and none of the others. Reactivity was limited to inclusions with the appearance of Lewy bodies, with the exception of one specimen in which intracellular reactivity was also observed in Nocardia-like structures. These results suggest a possible association between Nocardia and neurodegenerative disorders in which Lewy bodies are present. (C) 2003 Elsevier Inc. All rights reserved.