Antibody-directed extracellular proximity biotinylation reveals that Contactin-1 regulates axo-axonic innervation of axon initial segments.
Antibody-directed extracellular proximity biotinylation reveals that Contactin-1 regulates axo-axonic innervation of axon initial segments.
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DOI：
10.1038/s41467-023-42273-8
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发表时间：
2023-10-26
影响因子：
16.6
通讯作者：
Rasband, Matthew N
中科院分区：
文献类型：
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作者：
Ogawa, Yuki;Lim, Brian C;George, Shanu;Oses-Prieto, Juan A;Rasband, Joshua M;Eshed-Eisenbach, Yael;Hamdan, Hamdan;Nair, Supna;Boato, Francesco;Peles, Elior;Burlingame, Alma L;Van Aelst, Linda;Rasband, Matthew N
轴突起始段（AIS）细胞表面蛋白介导神经元中的关键生物学过程，包括动作电位起始和轴-轴突触形成。然而，很少有AIS细胞表面蛋白已被确定。在这里，我们使用抗体导向的邻近生物素化来定义与AIS细胞粘附分子Neurosfascin密切相关的细胞表面蛋白。为了确定所鉴定的蛋白质的分布，我们使用CRISPR介导的基因组编辑在内源性蛋白质中插入表位标签。我们确定Contactin-1（Cntn 1）作为AIS细胞表面蛋白。Cntn 1通过与Neurofascin和NrCAM的相互作用在AIS富集。我们进一步表明，Cntn 1有助于AIS细胞外基质的组装，并通过小脑中的抑制性篮状细胞和皮质中的抑制性枝状细胞来调节AIS轴-轴神经支配。在轴突起始段已经鉴定出很少的常驻细胞表面蛋白。在这里，Ogawa及其同事使用邻近标记和蛋白质组学来鉴定Contactin-1作为调节大脑布线的跨膜轴突起始片段蛋白。
Axon initial segment (AIS) cell surface proteins mediate key biological processes in neurons including action potential initiation and axo-axonic synapse formation. However, few AIS cell surface proteins have been identified. Here, we use antibody-directed proximity biotinylation to define the cell surface proteins in close proximity to the AIS cell adhesion molecule Neurofascin. To determine the distributions of the identified proteins, we use CRISPR-mediated genome editing for insertion of epitope tags in the endogenous proteins. We identify Contactin-1 (Cntn1) as an AIS cell surface protein. Cntn1 is enriched at the AIS through interactions with Neurofascin and NrCAM. We further show that Cntn1 contributes to assembly of the AIS extracellular matrix, and regulates AIS axo-axonic innervation by inhibitory basket cells in the cerebellum and inhibitory chandelier cells in the cortex. Few resident cell surface proteins have been identified at the axon initial segment. Here, Ogawa and colleagues use proximity labeling and proteomics to identify Contactin-1 as a transmembrane axon initial segment protein that regulates brain wiring.