No Dopamine Cell Loss or Changes in Cytoskeleton Function in Transgenic Mice Expressing Physiological Levels of Wild Type or G2019S Mutant LRRK2 and in Human Fibroblasts
No Dopamine Cell Loss or Changes in Cytoskeleton Function in Transgenic Mice Expressing Physiological Levels of Wild Type or G2019S Mutant LRRK2 and in Human Fibroblasts
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DOI：
10.1371/journal.pone.0118947
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发表时间：
2015-04-01
期刊：
影响因子：
3.7
通讯作者：
Biskup, Saskia
中科院分区：
文献类型：
--
作者：
Garcia-Miralles, Marta;Coomaraswamy, Janaky;Biskup, Saskia
LRRK2 基因内的突变已在帕金森病 (PD) 患者中被发现，并且与多种细胞通路的功能障碍有关。在这里，我们探讨了致病性突变和 LRRK2 激酶活性的抑制如何影响小鼠和人类细胞系统的细胞骨架动力学。我们生成并表征了表达人类野生型和 G2019S 突变体 LRRK2 生理水平的新型转基因小鼠模型。 12 个月大时，中脑多巴胺神经元未检测到神经元丢失或神经变性。来自转基因小鼠的出生后海马神经元显示，使用高内涵成像对数百个神经元自动采样的有关神经突生长的七个参数没有变化。用激酶抑制剂 LRRK2-IN-1 治疗后，神经突生长没有发生显着变化。在人类成纤维细胞中，我们分析了致病性 LRRK2 突变是否会改变细胞骨架功能，例如细胞粘附。为此，我们比较了来自六名携带三种不同致病性 LRRK2 突变之一的 PD 患者和四名年龄匹配的对照个体的人类皮肤成纤维细胞的粘附特征。突变的 LRRK2 变体以及 LRRK2 激酶活性的抑制并未显示培养的成纤维细胞中任何显着的细胞粘附差异。总之，我们在人和小鼠细胞系统中的结果表明野生型或突变型 LRRK2 的表达以及 LRRK2 激酶活性的抑制都不影响神经突复杂性和细胞粘附。
Mutations within the LRRK2 gene have been identified in Parkinson's disease (PD) patients and have been implicated in the dysfunction of several cellular pathways. Here, we explore how pathogenic mutations and the inhibition of LRRK2 kinase activity affect cytoskeleton dynamics in mouse and human cell systems. We generated and characterized a novel transgenic mouse model expressing physiological levels of human wild type and G2019S-mutant LRRK2. No neuronal loss or neurodegeneration was detected in midbrain dopamine neurons at the age of 12 months. Postnatal hippocampal neurons derived from transgenic mice showed no alterations in the seven parameters examined concerning neurite outgrowth sampled automatically on several hundred neurons using high content imaging. Treatment with the kinase inhibitor LRRK2-IN-1 resulted in no significant changes in the neurite outgrowth. In human fibroblasts we analyzed whether pathogenic LRRK2 mutations change cytoskeleton functions such as cell adhesion. To this end we compared the adhesion characteristics of human skin fibroblasts derived from six PD patients carrying one of three different pathogenic LRRK2 mutations and from four age-matched control individuals. The mutant LRRK2 variants as well as the inhibition of LRRK2 kinase activity did not reveal any significant cell adhesion differences in cultured fibroblasts. In summary, our results in both human and mouse cell systems suggest that neither the expression of wild type or mutant LRRK2, nor the inhibition of LRRK2 kinase activity affect neurite complexity and cellular adhesion.