Proteomic Analysis Reveals Sex-Specific Protein Degradation Targets in the Amygdala During Fear Memory Formation.
Proteomic Analysis Reveals Sex-Specific Protein Degradation Targets in the Amygdala During Fear Memory Formation.
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DOI：
10.3389/fnmol.2021.716284
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发表时间：
2021
影响因子：
4.8
通讯作者：
Jarome TJ
中科院分区：
文献类型：
--
作者：
Farrell K;Musaus M;Navabpour S;Martin K;Ray WK;Helm RF;Jarome TJ
泛素-蛋白酶体介导的蛋白质降解与杏仁核中恐惧记忆的形成有广泛的关系。然而，到目前为止，蛋白酶体的蛋白质靶点在很大程度上仍然未知，限制了我们对恐惧记忆形成中蛋白质降解的功能意义的理解。此外，两性之间的蛋白酶体是否靶向相似的蛋白质还有待探索。在这里，我们结合了降解特异性K48串联泛素结合实体（TUBE）与液相色谱质谱（LC/MS），以确定目标底物的蛋白质降解过程中的杏仁核的雄性和雌性大鼠的上下文恐惧条件反射。我们发现，男性（43名）和女性（77名）在恐惧条件反射后杏仁核中K48多聚泛素靶向蛋白质的总数方面存在差异。许多鉴定的蛋白质（106）在K48纯化的样品中在恐惧条件反射1小时后水平显著降低，表明由于学习导致底物的主动降解。有趣的是，只有3种蛋白质在性别之间重叠，这表明蛋白质降解过程的目标可能是性别特异性的。在女性中，许多蛋白质与K48纯化的样品中的丰度改变参与囊泡运输或与微管。相反，在男性中，参与细胞骨架，ATP合成和细胞信号传导的蛋白质被发现具有显着改变的丰度。只有一种蛋白质在性别之间的丰度有相反的方向变化，LENG 1，在男性中显着增强，而在女性中较低。这表明在恐惧记忆形成过程中，这种蛋白质在女性中的降解速度更快。有趣的是，星形胶质细胞结构的关键组成部分GFAP是男性和女性中K48多聚泛素化的目标，表明蛋白质降解可能发生在恐惧条件反射后的星形胶质细胞中。蛋白质印迹分析显示，在两种性别的恐惧条件反射后，这些靶底物的水平降低，证实了K48多聚泛素靶向这些蛋白质进行降解。总的来说，这项研究提供了强有力的证据表明，性别差异存在于杏仁核的降解过程中的蛋白质目标以下恐惧条件反射和关键信息如何泛素-蛋白酶体介导的蛋白质降解可能有助于恐惧记忆形成的大脑。
Ubiquitin-proteasome mediated protein degradation has been widely implicated in fear memory formation in the amygdala. However, to date, the protein targets of the proteasome remain largely unknown, limiting our understanding of the functional significance for protein degradation in fear memory formation. Additionally, whether similar proteins are targeted by the proteasome between sexes has yet to be explored. Here, we combined a degradation-specific K48 Tandem Ubiquitin Binding Entity (TUBE) with liquid chromatography mass spectrometry (LC/MS) to identify the target substrates of the protein degradation process in the amygdala of male and female rats following contextual fear conditioning. We found that males (43) and females (77) differed in the total number of proteins that had significant changes in K48 polyubiquitin targeting in the amygdala following fear conditioning. Many of the identified proteins (106) had significantly reduced levels in the K48-purified samples 1 h after fear conditioning, suggesting active degradation of the substrate due to learning. Interestingly, only 3 proteins overlapped between sexes, suggesting that targets of the protein degradation process may be sex-specific. In females, many proteins with altered abundance in the K48-purified samples were involved in vesicle transport or are associated with microtubules. Conversely, in males, proteins involved in the cytoskeleton, ATP synthesis and cell signaling were found to have significantly altered abundance. Only 1 protein had an opposite directional change in abundance between sexes, LENG1, which was significantly enhanced in males while lower in females. This suggests a more rapid degradation of this protein in females during fear memory formation. Interestingly, GFAP, a critical component of astrocyte structure, was a target of K48 polyubiquitination in both males and females, indicating that protein degradation is likely occurring in astrocytes following fear conditioning. Western blot assays revealed reduced levels of these target substrates following fear conditioning in both sexes, confirming that the K48 polyubiquitin was targeting these proteins for degradation. Collectively, this study provides strong evidence that sex differences exist in the protein targets of the degradation process in the amygdala following fear conditioning and critical information regarding how ubiquitin-proteasome mediated protein degradation may contribute to fear memory formation in the brain.
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影响因子：
4.8
作者：
Hegde AN;Haynes KA;Bach SV;Beckelman BC
通讯作者：
Beckelman BC
影响因子：
4.8
作者：
Dulka BN;Trask S;Helmstetter FJ
通讯作者：
Helmstetter FJ
影响因子：
2.7
作者：
Figueiredo, Luciana S.;Dornelles, Arethuza S.;Schroeder, Nadja
通讯作者：
Schroeder, Nadja
影响因子：
2.7
作者：
Furini, Cristiane R. G.;Myskiw, Jociane de C.;Izquierdo, Ivan
通讯作者：
Izquierdo, Ivan
影响因子：
2.7
作者：
Devulapalli R;Jones N;Farrell K;Musaus M;Kugler H;McFadden T;Orsi SA;Martin K;Nelsen J;Navabpour S;O'Donnell M;McCoig E;Jarome TJ
通讯作者：
Jarome TJ