Synaptic function is modulated by LRRK2 and glutamate release is increased in cortical neurons of G2019S LRRK2 knock-in mice.
Synaptic function is modulated by LRRK2 and glutamate release is increased in cortical neurons of G2019S LRRK2 knock-in mice.
复制标题
DOI：
10.3389/fncel.2014.00301
复制
发表时间：
2014
影响因子：
5.3
通讯作者：
Milnerwood AJ
中科院分区：
文献类型：
--
作者：
Beccano-Kelly DA;Kuhlmann N;Tatarnikov I;Volta M;Munsie LN;Chou P;Cao LP;Han H;Tapia L;Farrer MJ;Milnerwood AJ
富含亮氨酸重复激酶-2（LRRK 2）的突变导致家族性帕金森病，G2019 S突变在某些种族中占高达30%。尽管如此，LRRK 2的功能在很大程度上是不确定的，尽管有证据表明在磷酸化，蛋白质相互作用，自噬和内吞作用中的作用。新出现的报告将LRRK 2的丢失与突触传递的改变联系起来，但G2019 S突变对哺乳动物神经元突触释放的影响尚不清楚。为了评估已建立的神经元网络中的野生型和突变体LRRK 2，我们对LRRK 2敲除、野生型过表达和G2019 S敲入小鼠的>3周龄皮质培养物进行免疫细胞化学、电生理学和生物化学表征。在LRRK 2敲除细胞中，突触释放和突触数量大体正常，但观察到突触能活性离散降低和突触蛋白水平降低。相反，突触密度是适度的，但在野生型LRRK 2过表达的文化显着增加，虽然事件频率不是。在敲入培养物中，在突触密度没有任何变化的情况下，谷氨酸释放显著升高，表明G2019 S LRRK 2的生理水平提高了释放的可能性。一些突触前调节蛋白与LRRK 2相互作用，在敲入培养中以正常水平表达;然而，突触蛋白1磷酸化显着降低。因此，对突触前释放机制的扰动和升高的突触传递是LRRK 2 G2019 S的早期神经元效应。此外，敲入和过表达培养物的比较表明，G2019 S突变的一个拷贝具有比LRRK 2蛋白增加约3倍更显著的效果。Muc诱导的传输增加可能会向神经元生理学传递额外的应激源，最终可能导致帕金森病的发病机制。
Mutations in Leucine-Rich Repeat Kinase-2 (LRRK2) result in familial Parkinson's disease and the G2019S mutation alone accounts for up to 30% in some ethnicities. Despite this, the function of LRRK2 is largely undetermined although evidence suggests roles in phosphorylation, protein interactions, autophagy and endocytosis. Emerging reports link loss of LRRK2 to altered synaptic transmission, but the effects of the G2019S mutation upon synaptic release in mammalian neurons are unknown. To assess wild type and mutant LRRK2 in established neuronal networks, we conducted immunocytochemical, electrophysiological and biochemical characterization of >3 week old cortical cultures of LRRK2 knock-out, wild-type overexpressing and G2019S knock-in mice. Synaptic release and synapse numbers were grossly normal in LRRK2 knock-out cells, but discretely reduced glutamatergic activity and reduced synaptic protein levels were observed. Conversely, synapse density was modestly but significantly increased in wild-type LRRK2 overexpressing cultures although event frequency was not. In knock-in cultures, glutamate release was markedly elevated, in the absence of any change to synapse density, indicating that physiological levels of G2019S LRRK2 elevate probability of release. Several pre-synaptic regulatory proteins shown by others to interact with LRRK2 were expressed at normal levels in knock-in cultures; however, synapsin 1 phosphorylation was significantly reduced. Thus, perturbations to the pre-synaptic release machinery and elevated synaptic transmission are early neuronal effects of LRRK2 G2019S. Furthermore, the comparison of knock-in and overexpressing cultures suggests that one copy of the G2019S mutation has a more pronounced effect than an ~3-fold increase in LRRK2 protein. Mutant-induced increases in transmission may convey additional stressors to neuronal physiology that may eventually contribute to the pathogenesis of Parkinson's disease.
登录
查看更多内容
影响因子：
6.1
作者：
Melrose HL;Dächsel JC;Behrouz B;Lincoln SJ;Yue M;Hinkle KM;Kent CB;Korvatska E;Taylor JP;Witten L;Liang YQ;Beevers JE;Boules M;Dugger BN;Serna VA;Gaukhman A;Yu X;Castanedes-Casey M;Braithwaite AT;Ogholikhan S;Yu N;Bass D;Tyndall G;Schellenberg GD;Dickson DW;Janus C;Farrer MJ
通讯作者：
Farrer MJ
影响因子：
3.7
作者：
Kawakami F;Yabata T;Ohta E;Maekawa T;Shimada N;Suzuki M;Maruyama H;Ichikawa T;Obata F
通讯作者：
Obata F
影响因子：
5.3
作者：
Lin, Chin-Hsien;Tsai, Pei-I;Chien, Cheng-Ting
通讯作者：
Chien, Cheng-Ting
影响因子：
2.4
作者：
Biskup S;Moore DJ;Rea A;Lorenz-Deperieux B;Coombes CE;Dawson VL;Dawson TM;West AB
通讯作者：
West AB
影响因子：
5.7
作者：
Maffei, Arianna;Fontanini, Alfredo
通讯作者：
Fontanini, Alfredo