Mitogen-activated protein kinase regulates neurofilament axonal transport
Mitogen-activated protein kinase regulates neurofilament axonal transport
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DOI：
10.1242/jcs.01135
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发表时间：
2004-09-15
影响因子：
4
通讯作者：
Shea, TB
中科院分区：
文献类型：
--
作者：
Chan, WKH;Dickerson, A;Shea, TB
有丝分裂原活化蛋白激酶（MAP）激酶通过介导神经发生和神经元分化在神经系统的发育中起着关键作用。在这里，我们研究了p42/44 MAP激酶是否在轴突神经的轴突运输和神经丝（NFs）的组织中起作用。在分化的NB2a/d1神经母细胞瘤细胞中，显性阴性的p42/44 MAP激酶、抗MAP激酶反义寡核苷酸和MAP激酶抑制剂PD98059均能降低NF磷酸化表位，抑制gfp标记的NF亚基的顺行NF轴突转运。组成活性MAP激酶的表达和活性酶的细胞内传递增加了NF磷酸化表位和NF轴突转运。更长时间的PD98059治疗使NF转运从顺行转向逆行。PD98059不抑制整体轴突运输，也不损害整体轴突结构或组成。p38 MAP激酶抑制剂SB202190不抑制NF转运，而激酶抑制剂olomoucine同时抑制NF和线粒体转运。与野生型构建体相比，含有NF-H的NFs轴突运输受到PD98059的影响要小得多，其c端区域发生突变以模拟广泛的磷酸化。这些数据表明p42/44 MAP激酶通过NF c末端磷酸化调控NF顺行转运。因此，MAP激酶可能通过促进生长中的轴突神经突内NFs的积累来稳定发育中的轴突。
Mitogen-activated protein kinase (MAP) kinase plays a pivotal role in the development of the nervous system by mediating both neurogenesis and neuronal differentiation. Here we examined whether p42/44 MAP kinase plays a role in axonal transport and the organization of neurofilaments (NFs) in axonal neurites. Dominant-negative p42/44 MAP kinase, anti-MAP kinase antisense oligonucleotides and the MAP kinase inhibitor PD98059 all reduced NF phospho-epitopes and inhibited anterograde NF axonal transport of GFP-tagged NF subunits in differentiated NB2a/d1 neuroblastoma cells. Expression of constitutively active MAP kinase and intracellular delivery of active enzyme increased NF phospho-epitopes and increased NF axonal transport. Longer treatment with PD98059 shifted NF transport from anterograde to retrograde. PD98059 did not inhibit overall axonal transport nor compromise overall axonal architecture or composition. The p38 MAP kinase inhibitor SB202190 did not inhibit NF transport whereas the kinase inhibitor olomoucine inhibited both NF and mitochondrial transport. Axonal transport of NFs containing NF-H whose C-terminal region was mutated to mimic extensive phosphorylation was substantially less affected by PD98059 compared to a wild-type construct. These data suggest that p42/44 MAP kinase regulates NF anterograde transport by NF C-terminal phosphorylation. MAP kinase may therefore stabilize developing axons by promoting the accumulation of NFs within growing axonal neurites.