A comparative study of Lrrk2 function in primary neuronal cultures.
A comparative study of Lrrk2 function in primary neuronal cultures.
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DOI：
10.1016/j.parkreldis.2010.08.018
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发表时间：
2010-12
影响因子：
4.1
通讯作者：
Farrer MJ
中科院分区：
文献类型：
--
作者：
Dächsel JC;Behrouz B;Yue M;Beevers JE;Melrose HL;Farrer MJ
探讨富亮氨酸重复蛋白激酶2(LRRK2；LRRK2)野生型、突变型和缺失在树突状神经元树枝形成中的作用。LRRK2突变被认为是帕金森氏病易感性的主要遗传决定因素，为此对LRRK2突变功能的细胞分析将有助于靶向分子治疗药物的开发。尽管有高水平的人BAC野生型和突变过表达基因(Y1699C和G2019S)、小鼠敲除和G2019S敲除动物模型，但在BAC LRRK2野生型动物培养物中的树突神经元树枝状分布(轴突长度、分支和每个细胞的突起数量)是定量的，尽管人类转基因表达水平很高。相反，尽管每个细胞的突起总数保持不变，但两个BAC突变过表达的原代神经元表现出显著减少的神经细胞突起和分支。在BAC突变小鼠的培养中观察到的突变特异性毒性功能获得可能被星形孢子素治疗部分挽救，星形孢子素是一种非特异性激酶抑制剂。相反，在缺乏内源性LRRK2表达的小鼠基因敲除小鼠的神经元培养中，神经元树枝状分布要广泛得多。在LRRK2 G2019S敲入小鼠中，可以说是最具生理学相关性的系统，神经分支没有受到损害。在原代培养中，LRRK2的过度表达是一个夸大的、尽管是突变型特有的结果，但神经性树枝形成的损害。所描述的表型和测定提供了一种开发治疗药物的手段，该治疗药物调节突变LRRK2所产生的毒性功能获得。
To assess the contribution of wild-type, mutant and loss of leucine-rich repeat kinase 2 (LRRK2; Lrrk2) on dendritic neuronal arborization. LRRK2 mutations are recognized as the major genetic determinant of susceptibility to Parkinson’s disease for which a cellular assay of Lrrk2 mutant function would facilitate the development of targeted molecular therapeutics Dendritic neuronal arborization (neurite length, branching and the number of processes per cell) was quantified in primary hippocampal and midbrain cultures derived from five lines of recombinant LRRK2 mice, including human BAC wild-type and mutant overexpressors (Y1699C and G2019S), murine knock-out and G2019S knock-in animals Neuronal arborization in cultures from BAC Lrrk2 wild-type animals is comparable to non-transgenic littermate controls, despite high levels of human transgene expression. In contrast, primary neurons from both BAC mutant overexpressors presented with significantly reduced neuritic outgrowth and branching, although the total number of processes per cell remained comparable. The mutant-specific toxic gain-of-function observed in cultures from BAC mutant mice may be partially rescued by staurosporine treatment, a non-specific kinase inhibitor. In contrast, neuronal arborization is far more extensive in neuronal cultures derived from murine knock-out mice that lack endogenous Lrrk2 expression. In Lrrk2 G2019S knock-in mice, arguably the most physiologically relevant system, neuritic arborization is not impaired. Impairment of neuritic arborization is an exaggerated, albeit mutant-specific, consequence of Lrrk2 over-expression in primary cultures. The phenotype and assay described provides a means to develop therapeutic agents that modulate the toxic gain-of-function conferred by mutant Lrrk2.
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影响因子：
25
作者：
Li, Yanping;Liu, Wencheng;Oo, Tinmarla F.;Wang, Lei;Tang, Yi;Jackson-Lewis, Vernice;Zhou, Chun;Geghman, Kindiya;Bogdanov, Mikhail;Przedborski, Serge;Beal, M. Flint;Burke, Robert E.;Li, Chenjian
通讯作者：
Li, Chenjian
DOI：
10.1016/j.bbrc.2009.08.163
发表时间：
2009-11-20
影响因子：
3.1
作者：
Greggio, Elisa;Taymans, Jean-Marc;Zhen, Eugene Yuejun;Ryder, John;Vancraenenbroeck, Renee;Beilina, Alexandra;Sun, Peng;Deng, Junpeng;Jaffe, Howard;Baekelandt, Veerle;Merchant, Kalpana;Cookson, Mark R.
通讯作者：
Cookson, Mark R.
影响因子：
11.2
作者：
Jacquier, Arnaud;Buhler, Emmanuelle;Haase, Georg
通讯作者：
Haase, Georg
影响因子：
4.8
作者：
Greggio, Elisa;Zambrano, Ibardo;Cookson, Mark R.
通讯作者：
Cookson, Mark R.
DOI：
10.1016/j.bbrc.2007.10.198
发表时间：
2008-01-11
影响因子：
3.1
作者：
Temporin, Ko;Tanaka, Hiroyuki;Yoshikawa, Hideki
通讯作者：
Yoshikawa, Hideki