Carboxypeptidase E cytoplasmic tail mediates localization of synaptic vesicles to the pre-active zone in hypothalamic pre-synaptic terminals.
Carboxypeptidase E cytoplasmic tail mediates localization of synaptic vesicles to the pre-active zone in hypothalamic pre-synaptic terminals.
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DOI：
10.1111/j.1471-4159.2010.06820.x
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发表时间：
2010-08
影响因子：
4.7
通讯作者：
Loh YP
中科院分区：
文献类型：
--
作者：
Lou H;Park JJ;Cawley NX;Sarcon A;Sun L;Adams T;Loh YP
突触小泡 (SV) 如何定位到神经末梢的前活动区（活动区下方 5-200 nm）（可能代表慢反应 SV 库）尚不完全清楚。电镜显示，与野生型小鼠相比，羧肽酶 E 敲除 (CPE-KO) 小鼠下丘脑神经元中位于前活性区的 SV 数量显着减少。此外，我们发现 CPE-KO 小鼠下丘脑胚胎神经元 K+ 刺激的谷氨酸分泌受到损害。生化研究表明，来自野生型小鼠下丘脑的 SV 和来自 PC12 细胞的突触样微泡 (SLMV) 含有跨膜形式的 CPE，其胞质尾部 (CPEC10) 可能参与突触功能。酵母双杂交和下拉实验表明，CPE 胞质尾部与 γ-内收蛋白相互作用，γ-内收蛋白与神经末梢富集的肌动蛋白结合。使用 PC12 细胞作为模型的 TIRF 显微镜显示，GFP-CPEC15 的表达降低了在亚质膜下 200 nm 区域（TIRF 区）积累的含有 SLMV 的突触素-mRFP 的稳态水平。我们的研究结果确定了 CPE 细胞质尾部是下丘脑神经元富含肌动蛋白的前活性区和 PC12 细胞的 TIRF 区中 SV 定位的新介质。
How synaptic vesicles (SVs) are localized to the pre-active zone (5-200 nm beneath the active zone) in the nerve terminal, which may represent the slow response SV pool, is not fully understood. Electron microscopy revealed the number of SVs located in the pre-active zone, was significantly decreased in hypothalamic neurons of Carboxypeptidase E knockout (CPE-KO) mice compared to wild type mice. Additionally, we found K+-stimulated glutamate secretion from hypothalamic embryonic neurons was impaired in CPE-KO mice. Biochemical studies indicate that SVs from the hypothalamus of wild type mice and synaptic-like microvesicles (SLMVs) from PC12 cells contain a transmembrane form of CPE, with a cytoplasmic tail (CPEC10), maybe, involved in synaptic function. Yeast two-hybrid and pull-down experiments showed that the CPE cytoplasmic tail interacted with γ-adducin, which binds actin enriched at the nerve terminal. TIRF microscopy using PC12 cells as a model showed that expression of GFP-CPEC15 reduced the steady-state level of synaptophysin-mRFP containing SLMVs accumulated in the area within 200 nm under the sub-plasma membrane (TIRF zone). Our findings identify the CPE cytoplasmic tail, as a new mediator for the localization of SVs in the actin-rich pre-active zone in hypothalamic neurons and the TIRF zone of PC12 cells.
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DOI：
10.1523/jneurosci.0924-08.2008
发表时间：
2008-10-22
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Gitler D;Cheng Q;Greengard P;Augustine GJ
通讯作者：
Augustine GJ
影响因子：
--
作者：
Park, Joshua J.;Cawley, Niamh X.;Loh, Y. Peng
通讯作者：
Loh, Y. Peng
影响因子：
64.5
作者：
Cool, DR;Normant, E;Loh, YP
通讯作者：
Loh, YP
影响因子：
5.3
作者：
Siksou, Lea;Rostaing, Philippe;Marty, Serge
通讯作者：
Marty, Serge
影响因子：
64.8
作者：
Steyer, JA;Horstmann, H;Almers, W
通讯作者：
Almers, W