Study of protein and RNA in dendritic spines using multi-isotope imaging mass spectrometry (MIMS).
Study of protein and RNA in dendritic spines using multi-isotope imaging mass spectrometry (MIMS).
复制标题
DOI：
10.1002/sia.5617
复制
发表时间：
2014-11-01
期刊：
影响因子：
--
通讯作者：
Lechene C
中科院分区：
文献类型：
--
作者：
Brismar H;Aperia A;Westin L;Moy J;Wang M;Guillermier C;Poczatek C;Lechene C
关于神经元蛋白质合成的经典观点是，蛋白质在细胞体中产生，然后被运送到它们在树突和树突棘中的功能位点。然而，间接证据表明，蛋白质合成可以直接发生在远端树突，远离细胞体。我们正在开发在培养的神经元中使用15n -尿苷和18O-或13c -亮氨酸脉冲追踪对RNA和蛋白质进行双重标记的方案，以鉴定和定位蛋白质合成和新合成蛋白质的命运。初步实验表明，RNA和新合成的蛋白质在树突中的离散定位，接近树突棘。我们第一次直接成像并测量了亚细胞水平的蛋白质在神经元树突中的产生，靠近功能部位，树突棘。这将为研究健康和疾病中的神经生长和突触可塑性开辟一条强有力的途径。
The classical view of neuronal protein synthesis is that proteins are made in the cell body and then transported to their functional sites in the dendrites and the dendritic spines. Indirect evidence, however, suggests that protein synthesis can directly occur in the distal dendrites, far from the cell body. We are developing protocols for dual labeling of RNA and proteins using 15N-uridine and 18O- or 13C-leucine pulse chase in cultured neurons to identify and localize both protein synthesis and fate of newly synthesized proteins. Pilot experiments show discrete localization of both RNA and newly synthesized proteins in dendrites, close to dendritic spines. We have for the first time directly imaged and measured the production of proteins at the subcellular level in the neuronal dendrites, close to the functional sites, the dendritic spines. This will open a powerful way to study neural growth and synapse plasticity in health and disease.