Neuronal NADPH oxidase 2 regulates growth cone guidance downstream of slit2/robo2.
Neuronal NADPH oxidase 2 regulates growth cone guidance downstream of slit2/robo2.
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DOI：
10.1002/dneu.22791
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发表时间：
2021-01
影响因子：
3
通讯作者：
Suter DM
中科院分区：
文献类型：
--
作者：
Terzi A;Roeder H;Weaver CJ;Suter DM
NADPH 氧化酶 (Nox) 是膜结合的多亚基蛋白复合物，产生调节许多细胞过程的活性氧 (ROS)。新的证据表明，Nox 衍生的 ROS 还控制神经元发育和轴突生长。然而，Nox 是否作用于轴突生长和引导信号受体的下游目前尚不清楚。为了回答这个问题，我们培养了来自斑马鱼胚胎的视网膜神经节细胞 (RGC)，并将这些神经元暴露于 netrin-1、slit2 和脑源性神经营养因子 (BDNF)。为了测试 Nox 在线索介导的生长和引导中的作用，我们要么通过药理学抑制 Nox，要么研究缺乏 Nox2 的突变鱼的神经元。我们发现，slit2 介导的生长锥塌陷和轴突回缩可通过 Nox 抑制而消除。虽然我们没有看到 BDNF 或 netrin-1 对生长速率的影响，但在 netrin-1 存在的情况下，生长会因 Nox 抑制而降低。此外，当体外抑制Nox时，分别响应BDNF、netrin-1和slit2梯度的吸引和排斥生长锥转动被消除。 ROS 生物传感器成像显示，slit2 处理通过涉及 Nox2 激活的机制增加了生长锥过氧化氢水平。我们还研究了体内Nox2 和slit2/Robo2 信号传导之间可能的关系。与个体杂合子相比，astray/nox2双杂合子幼虫表现出顶盖神经支配面积减少，表明Nox2和Robo2都是建立视网膜顶盖连接所必需的。我们的结果提供了证据，证明 Nox2 通过介导斑马鱼 RGC 神经元的生长和指导来在 slim2/robo2 下游发挥作用。
NADPH oxidases (Nox) are membrane-bound multi-subunit protein complexes producing reactive oxygen species (ROS) that regulate many cellular processes. Emerging evidence suggests that Nox-derived ROS also control neuronal development and axonal outgrowth. However, whether Nox act downstream of receptors for axonal growth and guidance cues is presently unknown. To answer this question, we cultured retinal ganglion cells (RGCs) derived from zebrafish embryos and exposed these neurons to netrin-1, slit2, and brain-derived neurotrophic factor (BDNF). To test the role of Nox in cue-mediated growth and guidance, we either pharmacologically inhibited Nox or investigated neurons from mutant fish that are deficient in Nox2. We found that slit2-mediated growth cone collapse, and axonal retraction was eliminated by Nox inhibition. Though we did not see an effect of either BDNF or netrin-1 on growth rates, growth in the presence of netrin-1 was reduced by Nox inhibition. Furthermore, attractive and repulsive growth cone turning in response to gradients of BDNF, netrin-1, and slit2, respectively, were eliminated when Nox was inhibited in vitro. ROS biosensor imaging showed that slit2 treatment increased growth cone hydrogen peroxide levels via mechanisms involving Nox2 activation. We also investigated the possible relationship between Nox2 and slit2/Robo2 signaling in vivo. astray/nox2 double heterozygote larvae exhibited decreased area of tectal innervation as compared to individual heterozygotes, suggesting both Nox2 and Robo2 are required for establishment of retinotectal connections. Our results provide evidence that Nox2 acts downstream of slit2/robo2 by mediating growth and guidance of developing zebrafish RGC neurons.
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影响因子：
2.7
作者：
Cao, Xian;Demel, Stacie L.;Quinn, Mark T.;Galligan, James J.;Kreulen, David
通讯作者：
Kreulen, David
影响因子：
4.8
作者：
Bienert, Gerd P.;Moller, Anders L. B.;Jahn, Thomas P.
通讯作者：
Jahn, Thomas P.
影响因子：
3.3
作者：
Anderson, Tom W.;Vaughan, Andrew N.;Cramer, Louise P.
通讯作者：
Cramer, Louise P.
影响因子：
16.2
作者：
Deiner, MS;Kennedy, TE;Sretavan, DW
通讯作者：
Sretavan, DW
DOI：
10.1083/jcb.201102095
发表时间：
2011-07-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Finkel T
通讯作者：
Finkel T