A novel choline cotransporter sequestration compartment in cholinergic neurons revealed by selective endosomal ablation.
A novel choline cotransporter sequestration compartment in cholinergic neurons revealed by selective endosomal ablation.
复制标题
通过选择性内体消融揭示胆碱能神经元中新型胆碱协同转运蛋白隔离室。
DOI：
10.1111/j.1471-4159.2009.06543.x
复制
发表时间：
2010
影响因子：
4.7
通讯作者：
Townsel,JamesG
中科院分区：
文献类型：
--
作者：
Ivy,MichaelT;Newkirk,RobertF;Wang,Yilun;Townsel,JamesG
神经化学杂志（2010）112，1295- 1304.摘要钠依赖性高亲和力胆碱转运蛋白-胆碱协同转运蛋白-（ChCoT，aka：cho-1，CHT 1，CHT）在质膜和细胞质区室之间经历组成性和受调节的运输。这种贩运的途径和管理机制尚不清楚。我们在此报告涉及选择性内体消融的研究，以进一步了解ChCoT的贩运。早期分选和再循环内体的选择性消融导致[3 H]胆碱摄取减少约75%，[3 H]hemicholinium-3结合减少约70%。蛋白质印迹分析表明，消融产生了类似的减少ChCoTs在质膜亚细胞组分。该损失的时间范围约为2小时，这已被证明是该组织中ChCoT的组成性循环时间。 消融似乎依赖于转铁蛋白缀合的辣根过氧化物酶的细胞内循环和转铁蛋白缀合的辣根过氧化物酶在早期内体中的选择性沉积，分选和再循环。烧蚀的脑切片保留了其通过调节运输ChCoTs到质膜来招募的能力。这种ChCoTs的募集表明可募集的隔室与早期内体不同。这将是必要的，做进一步的研究，以确定新的螯合隔室支持ChCoT调节贩运。
J. Neurochem.(2010)112, 1295–1304.AbstractThe sodium‐dependent, high affinity choline transporter – choline cotransporter – (ChCoT, aka: cho‐1, CHT1, CHT) undergoes constitutive and regulated trafficking between the plasma membrane and cytoplasmic compartments. The pathways and regulatory mechanisms of this trafficking are not well understood. We report herein studies involving selective endosomal ablation to further our understanding of the trafficking of the ChCoT. Selective ablation of early sorting and recycling endosomes resulted in a decrease of ∼75% of [3H]choline uptake and ∼70% of [3H]hemicholinium‐3 binding. Western blot analysis showed that ablation produced a similar decrease in ChCoTs in the plasma membrane subcellular fraction. The time frame for this loss was approximately 2 h which has been shown to be the constitutive cycling time for ChCoTs in this tissue. Ablation appears to be dependent on the intracellular cycling of transferrin‐conjugated horseradish peroxidase and the selective deposition of transferrin‐conjugated horseradish peroxidase in early endosomes, both sorting and recycling. Ablated brain slices retained their capacity to recruit via regulated trafficking ChCoTs to the plasma membrane. This recruitment of ChCoTs suggests that the recruitable compartment is distinct from the early endosomes. It will be necessary to do further studies to identify the novel sequestration compartment supportive of the ChCoT regulated trafficking.