Intracellular injection of phospholipids directly alters exocytosis and the fraction of chemical release in chromaffin cells as measured by nano-electrochemistry.
Intracellular injection of phospholipids directly alters exocytosis and the fraction of chemical release in chromaffin cells as measured by nano-electrochemistry.
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DOI：
10.1039/d0sc03683h
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发表时间：
2020-10-06
期刊：
影响因子：
8.4
通讯作者：
Ewing AG
中科院分区：
文献类型：
--
作者：
Aref M;Ranjbari E;Romiani A;Ewing AG
使用纳米注射方法，我们引入了磷脂具有不同的内在几何形状到单个分泌细胞，并使用单细胞安培法（SCA）和细胞内囊泡影响电化学细胞仪（IVIEC）与纳米尖端电极监测胞内孵育的影响上的胞吐过程和囊泡存储。结合工具，这项工作提供了新的信息，以了解细胞内膜脂质工程的影响，胞吐释放，囊泡内容物和化学释放的分数。我们还评估了膜脂质改变对分离的囊泡的儿茶酚胺存储的影响，通过实施另一种安培技术，囊泡影响电化学细胞术（VIEC），细胞外。胞吐分析表明，磷脂酰胆碱和溶血磷脂酰胆碱的细胞内纳米注射减少释放的儿茶酚胺的数量，而磷脂酰乙醇胺显示相反的效果。这些观察结果支持了新出现的假设，脂质曲率导致膜重塑通过分泌途径，也提供了新的证据的关键作用的脂质定位在调节释放过程。有趣的是，IVIEC的数据意味着，总囊泡含量也受到原位补充的细胞与一些脂质，而相应的VIEC结果表明，在孤立的囊泡中的神经递质含量不受影响，通过改变囊泡膜脂质。这表明细胞内磷脂的干预对囊泡释放的细胞机制而不是囊泡结构具有影响，并导致有点令人惊讶的结论，即调节释放对囊泡结构具有直接影响，这可能是由于囊泡在胞吐期间再次打开和关闭。这些发现可能导致一种新的调控机制的胞吐或突触强度的基础上跨细胞膜的脂质异质性。安培法和细胞内囊泡影响电化学细胞术与nanotip电极被用来监测纳米注射不同几何形状的磷脂到分泌细胞后的胞吐和囊泡储存的影响。
Using a nano-injection method, we introduced phospholipids having different intrinsic geometries into single secretory cells and used single cell amperometry (SCA) and intracellular vesicle impact electrochemical cytometry (IVIEC) with nanotip electrodes to monitor the effects of intracellular incubation on the exocytosis process and vesicular storage. Combining tools, this work provides new information to understand the impact of intracellular membrane lipid engineering on exocytotic release, vesicular content and fraction of chemical release. We also assessed the effect of membrane lipid alteration on catecholamine storage of isolated vesicles by implementing another amperometric technique, vesicle impact electrochemical cytometry (VIEC), outside the cell. Exocytosis analysis reveals that the intracellular nano-injection of phosphatidylcholine and lysophosphatidylcholine decreases the number of released catecholamines, whereas phosphatidylethanolamine shows the opposite effect. These observations support the emerging hypothesis that lipid curvature results in membrane remodeling through secretory pathways, and also provide new evidence for a critical role of the lipid localization in modulating the release process. Interestingly, the IVIEC data imply that total vesicular content is also affected by in situ supplementation of the cells with some lipids, while, the corresponding VIEC results show that the neurotransmitter content in isolated vesicles is not affected by altering the vesicle membrane lipids. This suggests that the intervention of phospholipids inside the cell has its effect on the cellular machinery for vesicle release rather than vesicle structure, and leads to the somewhat surprising conclusion that modulating release has a direct effect on vesicle structure, which is likely due to the vesicles opening and closing again during exocytosis. These findings could lead to a novel regulatory mechanism for the exocytotic or synaptic strength based on lipid heterogeneity across the cell membrane. Amperometry and intracellular vesicle impact electrochemical cytometry with nanotip electrodes were used to monitor the effects on exocytosis and vesicular storage after nano-injection of phospholipids with different geometries into secretory cells.
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DOI：
10.1002/anie.201504839
发表时间：
2015-10-05
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Li X;Majdi S;Dunevall J;Fathali H;Ewing AG
通讯作者：
Ewing AG
影响因子：
13.3
作者：
Wimalasena, Kandatege
通讯作者：
Wimalasena, Kandatege
影响因子：
25
作者：
Choi, S;Klingauf, J;Tsien, RW
通讯作者：
Tsien, RW
影响因子：
2.9
作者：
Uchiyama, Yoshiko;Maxson, Marc M.;Ewing, Andrew G.
通讯作者：
Ewing, Andrew G.
影响因子：
5
作者：
Majdi, Soodabeh;Larsson, Anna;Ewing, Andrew G.
通讯作者：
Ewing, Andrew G.