Distinct dendritic spine and nuclear phases of calcineurin activation after exposure to amyloid-β revealed by a novel fluorescence resonance energy transfer assay.
Distinct dendritic spine and nuclear phases of calcineurin activation after exposure to amyloid-β revealed by a novel fluorescence resonance energy transfer assay.
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DOI：
10.1523/jneurosci.0227-12.2012
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发表时间：
2012-04-11
期刊：
影响因子：
--
通讯作者：
Hyman BT
中科院分区：
文献类型：
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作者：
Wu HY;Hudry E;Hashimoto T;Uemura K;Fan ZY;Berezovska O;Grosskreutz CL;Bacskai BJ;Hyman BT
钙调神经磷酸酶（calcalineurin, CaN）的激活对低聚淀粉样蛋白β (Aβ)对脊柱形态的调节以及正常记忆中的突触可塑性至关重要，但目前还没有技术可以监测CaN活性的时空模式。在这里，我们使用光谱荧光共振能量转移（FRET）方法实时监测CaN激活动态与亚细胞分辨率。将Tg2576小鼠转基因神经元或人AD大脑中提取的寡聚物a β应用于野生型小鼠原代皮质神经元时，我们观察到CaN在几分钟内的动态激活过程，首先在树突棘中激活，然后在细胞质中激活，几小时后在细胞核中激活。CaN在脊髓中的激活导致脊髓和突触后蛋白快速但可逆的形态学改变；长时间暴露会导致NFAT移位到细胞核和脊柱损伤。这些结果为理解钙调磷酸酶在与AD发病机制相关的突触改变中的作用提供了一个框架。
Calcineurin (CaN) activation is critically involved in the regulation of spine morphology in response to oligomeric amyloid β (Aβ) as well as in synaptic plasticity in normal memory, but no existing techniques can monitor the spatiotemporal pattern of CaN activity. Here we use a spectral Fluorescence Resonance Energy Transfer (FRET) approach to monitor CaN activation dynamics in real time with subcellular resolution. When oligomeric Aβ derived from Tg2576 murine transgenic neurons or human AD brains were applied to wild-type murine primary cortical neurons, we observe a dynamic progression of CaN activation within minutes, first in dendritic spines, then in the cytoplasm and, in hours, in the nucleus. CaN activation in spines leads to rapid but reversible morphological changes in spines and in postsynaptic proteins; longer exposure leads to NFAT translocation to the nucleus and frank spine loss. These results provide a framework for understanding calcineurin’s role in synaptic alterations associated with AD pathogenesis.