Spatial distribution of DARPP-32 in dendritic spines.
Spatial distribution of DARPP-32 in dendritic spines.
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DOI：
10.1371/journal.pone.0075155
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Brismar H
中科院分区：
文献类型：
--
作者：
Blom H;Rönnlund D;Scott L;Westin L;Widengren J;Aperia A;Brismar H
磷蛋白DARPP-32（多巴胺和环腺苷3 ′，5 ′-单磷酸调节磷蛋白，32 kDa）是新纹状体突触后信号分子调节的重要组成部分。尽管这种磷蛋白很重要，但人们对DARPP-32的纳米级分布知之甚少。在这项研究中，我们应用超分辨率受激发射损耗显微镜（STED），以评估DARPP-32在纹状体神经元的表达和分布。纹状体神经元的原代培养物用Alexa-594免疫荧光标记DARPP-32，用atto-647 N免疫荧光标记多巴胺D1受体（D1 R）。双色STED显微镜显示DARPP-32和D1 R在脊柱结构中的离散定位，在头部和颈部都有聚集分布。解剖的脊柱结构显示DARPP-32信号很少与D1 R信号重叠。D1 R受体以“聚集”的方式主要位于脊柱头部，并在一定程度上位于颈部，而DARPP-32形成跨越整个脊柱结构的几个相邻的小纳米簇。DARPP-32簇的平均尺寸为52 +/- 6 nm，接近显微镜的分辨率极限，对应于一些单个磷蛋白免疫复合物的物理尺寸。使用超分辨率显微镜的突触蛋白的解剖提供了可能性，以更好地揭示详细的生物相关信息，衍射限制显微镜相比。在这项工作中，DARPP-32磷蛋白的解剖突触后拓扑结构为树突棘中的区室化和限制性分布提供了强有力的证据。蛋白质拓扑结构和磷蛋白相对低的拷贝数提供了DARPP-32微调突触信号调节的可能性的概念，这应该对表达它的神经元回路的性能产生影响。
The phosphoprotein DARPP-32 (dopamine and cyclic adenosine 3´, 5´-monophosphate-regulated phosphoprotein, 32 kDa) is an important component in the molecular regulation of postsynaptic signaling in neostriatum. Despite the importance of this phosphoprotein, there is as yet little known about the nanoscale distribution of DARPP-32. In this study we applied superresolution stimulated emission depletion microscopy (STED) to assess the expression and distribution of DARPP-32 in striatal neurons. Primary culture of striatal neurons were immunofluorescently labeled for DARPP-32 with Alexa-594 and for the dopamine D1 receptor (D1R) with atto-647N. Dual-color STED microscopy revealed discrete localizations of DARPP-32 and D1R in the spine structure, with clustered distributions in both head and neck. Dissected spine structures reveal that the DARPP-32 signal rarely overlapped with the D1R signal. The D1R receptor is positioned in an “aggregated” manner primarily in the spine head and to some extent in the neck, while DARPP-32 forms several neighboring small nanoclusters spanning the whole spine structure. The DARPP-32 clusters have a mean size of 52 +/- 6 nm, which is close to the resolution limit of the microscope and corresponds to the physical size of a few individual phosphoprotein immunocomplexes. Dissection of synaptic proteins using superresolution microscopy gives possibilities to reveal in better detail biologically relevant information, as compared to diffraction-limited microscopy. In this work, the dissected postsynaptic topology of the DARPP-32 phosphoprotein provides strong evidence for a compartmentalized and confined distribution in dendritic spines. The protein topology and the relatively low copy number of phosphoprotein provides a conception of DARPP-32’s possibilities to fine-tune the regulation of synaptic signaling, which should have an impact on the performance of the neuronal circuits in which it is expressed.
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影响因子：
4.3
作者：
Fernandez E;Schiappa R;Girault JA;Le Novère N
通讯作者：
Le Novère N
影响因子：
2.5
作者：
OUIMET, CC;LAMANTIA, AS;GREENGARD, P
通讯作者：
GREENGARD, P
DOI：
10.1073/pnas.032654599
发表时间：
2002-02-05
影响因子：
11.1
作者：
Scott, L;Kruse, MS;Aperia, A
通讯作者：
Aperia, A
影响因子：
25
作者：
Aizman, O;Brismar, H;Aperia, A
通讯作者：
Aperia, A
DOI：
10.1007/bf01188438
发表时间：
1990-02-01
期刊：
JOURNAL OF NEUROCYTOLOGY
影响因子：
--
作者：
OUIMET, CC;GREENGARD, P
通讯作者：
GREENGARD, P