N-cadherin and Neuroligins Cooperate to Regulate Synapse Formation in Hippocampal Cultures
N-cadherin and Neuroligins Cooperate to Regulate Synapse Formation in Hippocampal Cultures
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DOI：
10.1074/jbc.m110.176305
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发表时间：
2011-01-07
影响因子：
4.8
通讯作者：
Bamji, Shernaz X.
中科院分区：
文献类型：
--
作者：
Aiga, Mytyl;Levinson, Joshua N.;Bamji, Shernaz X.
钙粘蛋白和神经连接蛋白（NLs）代表了两个家族的细胞粘附蛋白，这是建立突触连接在体外是必不可少的，但是，目前还不清楚这些蛋白质是否一致行动，以调节突触密度。在原代海马神经元中使用过表达和敲低分析的组合，我们证明了NL 1和N-钙粘蛋白通过共同的功能途径促进海马神经元突触的形成。N-cadherin和NL 1之间的空间关系的分析表明，在14天的体外培养，几乎一半的突触与这两种蛋白质，而只有一个子集的这些突触与N-cadherin或NL 1单独。这表明，NL 1和N-钙粘蛋白的空间分布的方式，使合作的突触。在年轻的文化，N-钙粘蛋白聚类及其与突触标记的关联之前的NL 1聚类。N-cadherin在该时间点的过表达增强NL 1聚集并增加突触密度。虽然N-钙粘蛋白是不足以提高NL 1集群和突触密度在更成熟的文化，敲低N-钙粘蛋白在稍后的时间点显着衰减NL 1集群和突触的密度。N-cadherin过表达可以部分挽救NL 1敲低细胞中的突触丢失，这可能是由于N-cadherin能够将NL 2募集到这些细胞中的突触能突触。我们证明，钙粘蛋白和神经胶质细胞可以采取一致行动，以调节突触的形成。
Cadherins and neuroligins (NLs) represent two families of cell adhesion proteins that are essential for the establishment of synaptic connections in vitro; however, it remains unclear whether these proteins act in concert to regulate synapse density. Using a combination of overexpression and knockdown analyses in primary hippocampal neurons, we demonstrate that NL1 and N-cadherin promote the formation of glutamatergic synapses through a common functional pathway. Analysis of the spatial relationship between N-cadherin and NL1 indicates that in 14-day in vitro cultures, almost half of glutamatergic synapses are associated with both proteins, whereas only a subset of these synapses are associated with N-cadherin or NL1 alone. This suggests that NL1 and N-cadherin are spatially distributed in a manner that enables cooperation at synapses. In young cultures, N-cadherin clustering and its association with synaptic markers precede the clustering of NL1. Overexpression of N-cadherin at this time point enhances NL1 clustering and increases synapse density. Although N-cadherin is not sufficient to enhance NL1 clustering and synapse density in more mature cultures, knockdown of N-cadherin at later time points significantly attenuates the density of NL1 clusters and synapses. N-cadherin overexpression can partially rescue synapse loss in NL1 knockdown cells, possibly due to the ability of N-cadherin to recruit NL2 to glutamatergic synapses in these cells. We demonstrate that cadherins and NLs can act in concert to regulate synapse formation.