Neurexin Ibeta and neuroligin are localized on opposite membranes in mature central synapses.

Neurexin Ibeta and neuroligin are localized on opposite membranes in mature central synapses.
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Neurexin Ibeta和Neuroligin位于成熟中央突触中的相对膜上。

DOI:
10.1111/j.1471-4159.2007.04918.x
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发表时间:
2007-12
影响因子:
4.7
通讯作者:
Ushkaryov YA
Ushkaryov YA
中科院分区:
医学2区
文献类型:
--
作者:
Berninghausen O;Rahman MA;Silva JP;Davletov B;Hopkins C;Ushkaryov YA

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突触发生需要在神经元细胞黏附受体之间形成跨突触复合体。在突触前和突触后的神经元中,异嗜性受体对,如Neuresin Iβ和Neurn Ligin,可以介导不同的细胞内信号并形成不同的细胞质支架,可能对突触发生特别重要。然而,神经尿素素和神经连接素的功能取决于它们在突触中的分布。神经连接素已被实验性地分配到突触后膜上,而神经尿素素的定位尚不清楚。为了研究神经尿素Iβ和神经连接蛋白在成熟大脑皮层突触中的亚细胞分布,我们建立了一种新的物理分离连接膜的方法,并用Western blotting直接分析它们。使用尿素和DTT,我们破坏了跨突触蛋白连接,而不溶解脂质相,并分离了突触前和突触后的膜。这些组分的纯度通过电子显微镜和多突触标记的Western blotting进行了验证。定量分析证实,神经连接素仅定位于突触后膜。我们还证明,Neuresin Iβ很大程度上(96%)是突触前的。因此,神经毒素Iβ和神经连接蛋白通常形成跨突触复合体,并能传递双向信号。
Synaptogenesis requires formation of trans-synaptic complexes between neuronal cell-adhesion receptors. Heterophilic receptor pairs, such as neurexin Iβ and neuroligin, can mediate distinct intracellular signals and form different cytoplasmic scaffolds in the pre- and postsynaptic neuron, and may be particularly important for synaptogenesis. However, the functions of neurexin and neuroligin depend on their distribution in the synapse. Neuroligin has been experimentally assigned to the postsynaptic membrane, while the localization of neurexin remains unclear. To study the subcellular distribution of neurexin Iβ and neuroligin in mature cerebrocortical synapses, we have developed a novel method for physical separation of junctional membranes and their direct analysis by western blotting. Using urea and DTT, we disrupted trans-synaptic protein links, without dissolving the lipid phase, and fractionated the pre- and postsynaptic membranes. The purity of these fractions was validated by electron microscopy and western blotting using multiple synaptic markers. A quantitative analysis has confirmed that neuroligin is localized strictly in the postsynaptic membrane. We have also demonstrated, that neurexin Iβ is largely (96%) presynaptic. Thus, neurexin Iβ and neuroligin normally form trans-synaptic complexes and can transduce bidirectional signals.
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