Regulated interactions of the norepineprhine transporter by the actin and microtubule cytoskeletons
Regulated interactions of the norepineprhine transporter by the actin and microtubule cytoskeletons
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DOI：
10.1111/j.1471-4159.2008.05258.x
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发表时间：
2008-06-01
影响因子：
4.7
通讯作者：
Sidhu, Anita
中科院分区：
文献类型：
--
作者：
Jeannotte, Alexis M.;Sidhu, Anita
肌动蛋白细胞骨架的一个作用是维持突触前末端的结构形态和活性。我们试图确定肌动蛋白细胞骨架是否在调节去甲肾上腺素转运蛋白（NET）和α -突触核蛋白（α -syn）之间的相互作用中起作用，这两种蛋白在突触前末端表达。在转染了0.5 μ g/mL或3 μ g/mL α - syn和1 μ g/mL NET DNA的细胞中，细胞松弛素D（一种肌动蛋白解聚剂）分别引起[H-3]-NE摄取的剂量依赖性减少和增加。NET、β -肌动蛋白和α - syn之间的蛋白相互作用以及表面转运蛋白的水平都被改变了。用细胞松弛素D处理初级脑干神经元和额叶皮质突触体分别使NET活性增加115%和28%。在α - syn浓度为0.5 μ g/mL的细胞中，肌动蛋白和微管解聚均未改变NET活性，但在α - syn浓度为3 μ g/mL的细胞和原代神经元中，[H-3]-NE摄取增加。这是第一次直接证明NET活性通过肌动蛋白调节，并通过与α - syn的相互作用进行调节。
One role of the actin cytoskeleton is to maintain the structural morphology and activity of the pre-synaptic terminal. We sought to determine if the actin cytoskeleton plays a role in regulating interactions between the norepinephrine transporter (NET) and alpha-Synuclein (alpha-Syn), two proteins expressed in the pre-synaptic terminal. In cells transfected with either 0.5 mu g/mL or 3 mu g/mL of alpha-Syn and 1 mu g/mL of NET DNA, treatment with cytochalasin D, an actin depolymerizing agent, caused a dose-dependent decrease and increase, respectively, in [H-3]-NE uptake. Protein interactions between NET, beta-actin, and alpha-Syn were modified, along with levels of surface transporters. Treatment of primary brainstem neurons and frontal cortex synaptosomes with cytochalasin D caused a 115% and 28% increase, respectively, in NET activity. Depolymerization of both actin and microtubules did not alter NET activity in cells with 0.5 mu g/mL alpha-Syn, but caused an increase in [H-3]-NE uptake in cells transfected with 3 mu g/mL of alpha-Syn and primary neurons. This is the first direct demonstration of NET activity being regulated via actin and modulated by interactions with alpha-Syn.