Presynaptic calcium channels and α3-integrins are complexed with synaptic cleft laminins, cytoskeletal elements and active zone components.

Presynaptic calcium channels and α3-integrins are complexed with synaptic cleft laminins, cytoskeletal elements and active zone components.
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DOI:
10.1111/j.1471-4159.2010.06965.x
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发表时间:
2010-11
影响因子:
4.7
通讯作者:
Sanes JR
Sanes JR
中科院分区:
医学2区
文献类型:
--
作者:
Carlson SS;Valdez G;Sanes JR

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在化学突触中,突触间隙成分与神经终末膜的成分相互作用以促进分化和调节功能。含有β2亚基的层粘连蛋白是关键的裂隙组分,它们部分通过结合突触前电压门控钙通道(Cavα)的成孔亚基而起作用。在这里,我们确定了Cavα相关的细胞内蛋白,可能耦合通道锚定组装或稳定的神经递质释放位点称为活动区。使用Cavα-抗体,我们从电鳐电器官突触中分离出一种蛋白复合物,这种蛋白复合物类似于神经肌肉接头,但更容易批量分离。我们鉴定了该复合物的10种组分:6种细胞骨架蛋白(α2/β2血影蛋白、网蛋白1、AHNAK/桥粒蛋白、肌营养不良蛋白和肌球蛋白1)、2种活性区组分(巴松管和短笛)、突触层粘连蛋白和钙通道β亚基。免疫细胞化学证实了这些蛋白质的电器官突触,和PCR分析揭示了它们的表达发育中的哺乳动物运动神经元。最后,我们表明,突触层粘连蛋白也相互作用的突触前整合素含有α3亚基。结合我们先前的发现,即一种独特的突触层粘连蛋白与神经末梢上的SV 2相互作用,我们的研究结果确定了突触间隙层粘连蛋白可以向神经末梢发送发育重要信号的三种途径。
At chemical synapses, synaptic cleft components interact with elements of the nerve terminal membrane to promote differentiation and regulate function. Laminins containing the β2 subunit are key cleft components, and they act in part by binding the pore-forming subunit of a presynaptic voltage-gated calcium channel (Cavα). Here, we identify Cavα-associated intracellular proteins that may couple channel-anchoring to assembly or stabilization of neurotransmitter release sites called active zones. Using Cavα–antibodies, we isolated a protein complex from Torpedo electric organ synapses, which resemble neuromuscular junctions but are easier to isolate in bulk. We identified 10 components of the complex: six cytoskeletal proteins (α2/β2 spectrins, plectin 1, AHNAK/desmoyokin, dystrophin, and myosin 1), two active zone components (bassoon and piccolo), synaptic laminin, and a calcium channel β subunit. Immunocytochemistry confirmed these proteins in electric organ synapses, and PCR analysis revealed their expression by developing mammalian motor neurons. Finally, we show that synaptic laminins also interact with presynaptic integrins containing the α3 subunit. Together with our previous finding that a distinct synaptic laminin interacts with SV2 on nerve terminals, our results identify three paths by which synaptic cleft laminins can send developmentally important signals to nerve terminals.
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