JNK1-Dependent Phosphorylation of GAP-43 Serine 142 is a Novel Molecular Marker for Axonal Growth
JNK1-Dependent Phosphorylation of GAP-43 Serine 142 is a Novel Molecular Marker for Axonal Growth
复制标题
DOI：
10.1007/s11064-022-03580-6
复制
发表时间：
2022-03-26
影响因子：
4.4
通讯作者：
Igarashi，Michihiro
中科院分区：
文献类型：
--
作者：
Okada，Masayasu;Kawagoe，Yosuke;Igarashi，Michihiro
哺乳动物轴突生长与轴突再生具有相似的机制。生长锥是参与这两个过程的重要结构，并且GAP-43（生长相关蛋白-43kDa）被认为是经典的分子标记。以前，我们使用生长锥磷酸化蛋白质组学来证明啮齿动物中GAP-43的S96和T172是被c-junN-末端蛋白激酶（JNK）磷酸化的高度磷酸化位点。我们还发现，磷酸化的（p）S96和pT 172抗体识别发育中的大脑和再生轴突在成人周围神经轴突的生长。在啮齿类动物中，S142是另一个假定的JNK依赖性磷酸化位点，其修饰频率低于S96和T172。在这里，我们使用pS142特异性抗体对该位点进行了表征。我们证实，pS142是通过共表达小鼠GAP-43和JNK 1检测到的。pS142抗体标记发育中的小鼠神经元中的生长锥和生长轴突。pS142在小鼠脑中持续到出生后至少9周。pS142抗体可以检测成年小鼠坐骨神经损伤后再生的轴突。氨基酸序列的比较表明啮齿动物S142对应于人S151，其被预测为MAPK家族的底物，该MAPK家族包括JNK。因此，我们证实了pS142抗体使用活化的JNK 1识别人磷酸化GAP-43，并且其在从人诱导多能细胞分化的神经元中的免疫染色模式与在小鼠中观察到的相似。这些结果表明，S142残基被JNK 1磷酸化，并且pS142抗体是啮齿动物和人类轴突生长的新的候选分子标记。
Mammalian axon growth has mechanistic similarities with axon regeneration. The growth cone is an important structure that is involved in both processes, and GAP-43 (growth associated protein-43 kDa) is believed to be the classical molecular marker. Previously, we used growth cone phosphoproteomics to demonstrate that S96 and T172 of GAP-43 in rodents are highly phosphorylated sites that are phosphorylated by c-junN-terminal protein kinase (JNK). We also revealed that phosphorylated (p)S96 and pT172 antibodies recognize growing axons in the developing brain and regenerating axons in adult peripheral nerves. In rodents, S142 is another putative JNK-dependent phosphorylation site that is modified at a lower frequency than S96 and T172. Here, we characterized this site using a pS142-specific antibody. We confirmed that pS142 was detected by co-expressing mouse GAP-43 and JNK1. pS142 antibody labeled growth cones and growing axons in developing mouse neurons. pS142 was sustained until at least nine weeks after birth in mouse brains. The pS142 antibody could detect regenerating axons following sciatic nerve injury in adult mice. Comparison of amino acid sequences indicated that rodent S142 corresponds to human S151, which is predicted to be a substrate of the MAPK family, which includes JNK. Thus, we confirmed that the pS142 antibody recognized human phospho-GAP-43 using activated JNK1, and also that its immunostaining pattern in neurons differentiated from human induced pluripotent cells was similar to those observed in mice. These results indicate that the S142 residue is phosphorylated by JNK1 and that the pS142 antibody is a new candidate molecular marker for axonal growth in both rodents and human.