Transfer of synaptic vesicle antigens to the presynaptic plasma membrane during exocytosis.
Transfer of synaptic vesicle antigens to the presynaptic plasma membrane during exocytosis.
复制标题
胞吐作用期间突触小泡抗原转移至突触前质膜。
DOI：
10.1073/pnas.78.2.1014
复制
发表时间：
1981
影响因子：
11.1
通讯作者：
Kelly,RB
中科院分区：
文献类型：
--
作者：
vonWedel,RJ;Carlson,SS;Kelly,RB
我们利用免疫荧光技术在静息和活动神经末梢的质膜上寻找突触囊泡抗原。从海洋电鱼(Narcine Brasilisis)的电器官中提纯胆碱能突触囊泡，制备兔抗血清。在青蛙神经肌肉标本的冰冻切片中，突触小泡的抗体被选择性地结合到神经末梢上。用荧光素标记的山羊抗兔抗体间接证明结合。由于它们的大小、形状和位置，以及它们与罗丹明结合的α-银环蛇毒素结合部位和具有乙酰胆碱酯酶活性的部位重合，结合抗血清的横截面结构被确定为神经末梢。据推测，切片使抗体能够接触到神经末梢内的结合部位。然而，当在切片前将抗突触小泡的抗体加入完整的神经肌肉标本的浴液中时，抗体结合是边缘的或检测不到的，这表明正常情况下很少有小泡抗原可在静息神经末梢的外表面获得。当加入1 mM LaCl3刺激完整的制剂释放囊泡性乙酰胆碱时，抗体与完整神经末梢的结合变得显著。这些结果表明，突触囊泡膜和突触终末质膜的组成不同。它们还进一步支持了神经递质释放的胞吐假说，该假说预测，在刺激过程中，当小泡与质膜融合时，小泡标志物应该暴露在神经末梢的外部。
We have utilized immunofluorescence techniques to look for synaptic vesicle antigens on the plasma membrane of resting and active nerve terminals. Rabbit antiserum was raised against purified cholinergic synaptic vesicles from the electric organ of Narcine brasiliensis, a marine electric ray. Antibodies to synaptic vesicles were shown to bind selectively to nerve terminals in cryostat sections of frog nerve-muscle preparations. Binding was demonstrated indirectly by using fluorescein-labeled goat anti-rabbit antibodies. Structures in cross sections that bound antiserum were identified as nerve terminals because of their size, shape, and position and because they coincided with sites that bound rhodamine-conjugated alpha-bungarotoxin and had acetylcholine esterase activity. Presumably, sectioning gave antibodies access to binding sites within the nerve terminal. However, when antibodies to synaptic vesicles were added to the bathing medium of intact neuromuscular preparations prior to sectioning, antibody binding was marginal or undetectable, suggesting that few vesicle antigens were normally accessible on the outer surface of resting nerve terminals. When intact preparations were stimulated to release their vesicular acetylcholine by the addition of 1 mM LaCl3, antibody binding to the intact nerve terminals became striking. These findings suggest that the synaptic vesicle membrane and the synaptic terminal plasma membrane differ in composition. They also provide further support for the exocytotic hypothesis of neurotransmitter release, which predicts that vesicle markers should be exposed on the outside of nerve terminals when vesicles fuse with the plasma membrane during stimulation.
胆碱能机制和精神药理学
DOI：
--
发表时间：
1978
期刊：
Advances in behavioral biology
影响因子：
--
作者：
D. Jenden
通讯作者：
D. Jenden