BONLAC: A combinatorial proteomic technique to measure stimulus-induced translational profiles in brain slices.
BONLAC: A combinatorial proteomic technique to measure stimulus-induced translational profiles in brain slices.
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DOI：
10.1016/j.neuropharm.2015.07.017
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发表时间：
2016-01
影响因子：
4.7
通讯作者：
Klann E
中科院分区：
文献类型：
--
作者：
Bowling H;Bhattacharya A;Zhang G;Lebowitz JZ;Alam D;Smith PT;Kirshenbaum K;Neubert TA;Vogel C;Chao MV;Klann E
刺激触发的蛋白质合成对于大脑健康和功能至关重要。然而，由于技术障碍，从头上的神经元翻译主要在培养的细胞中进行了研究，而电生理和电路分析经常在脑切片中进行。这两个实验系统的不同特性造成了有关刺激诱导的成熟电路中新蛋白表达改变的信息差距。为了解决这个问题，我们将两种现有技术（Boncat和Silac）调整为Bonlac的组合蛋白质组学技术，用于急性成人海马片。我们使用BDNF诱导的蛋白质合成作为概念证明，我们发现在神经传递，运输和阳离子结合中与在培养神经元中相似筛查中发现的蛋白质表达发生了变化。我们的结果表明，培养的神经元和切片之间的重要差异，并表明BONLAC可用于剖析成人电路中突触事件的蛋白质组学变化。
Stimulus-triggered protein synthesis is critical for brain health and function. However, due to technical hurdles, de novo neuronal translation is predominantly studied in cultured cells, whereas electrophysiological and circuit analyses often are performed in brain slices. The different properties of these two experimental systems create an information gap about stimulus-induced alterations in the expression of new proteins in mature circuits. To address this, we adapted two existing techniques, BONCAT and SILAC, to a combined proteomic technique, BONLAC, for use in acute adult hippocampal slices. Using BDNF-induced protein synthesis as a proof of concept, we found alterations in expression of proteins involved in neurotransmission, trafficking, and cation binding that differed from those found in a similar screen in cultured neurons. Our results indicate important differences between cultured neurons and slices, and suggest that BONLAC could be used to dissect proteomic changes underlying synaptic events in adult circuits.