Cell survival controlled by lens-derived Sema3A-Nrp1 is vital on caffeine-suppressed corneal innervation during chick organogenesis
Cell survival controlled by lens-derived Sema3A-Nrp1 is vital on caffeine-suppressed corneal innervation during chick organogenesis
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由晶状体衍生的 Sema3A-Nrp1 控制的细胞存活对于雏鸡器官发生过程中咖啡因抑制的角膜神经支配至关重要
DOI：
10.1002/jcp.27671
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发表时间：
2019
影响因子：
5.6
通讯作者：
Yang Xuesong
中科院分区：
文献类型：
--
作者：
Wang Guang;Jiang Ling Min;Tan Bao Yi;Li Pei Zhi;Zhang Pei Ling;Zhang Yu;Cheng Xin;Ma Zheng Lai;Li Zhi Jie;Br;Saberi Beate;Yang Xuesong
在这项研究中，我们研究了咖啡因过度暴露对早期鸡胚角膜神经支配的影响。咖啡因给药通过影响三叉神经发育限制角膜神经支配。磷酸组蛋白3（pHIS 3）和C-caspase 3的免疫组织化学显示，细胞存活受到咖啡因给药的抑制。针对semaphorin 3A（Sema 3A）和neuropilin-1（Nrp 1）的整体原位杂交显示，咖啡因和2，2 ′-偶氮二（2-甲基丙脒）二盐酸盐（AAPH，一种自由基发生剂）给药均上调Sema 3A和Nrp 1的表达。接下来，我们证明了在发育中的鸡胚胎中的透镜消融显著影响NF标记的眼周神经束和中央眼区域的神经支配。随后，我们使用神经母细胞瘤细胞系在体外研究Sema 3A-Nrp 1信号传导是否对咖啡因抑制的神经元存活发挥关键作用。通过转染Sema 3A-siRNA敲低Sema 3A显著降低了细胞对咖啡因给药的反应性以及细胞凋亡。我们认为Sema 3A-Nrp 1信号通过Slit 2-Robo 1和Ephb 2调节Trp 53和Cdkn 1a。综上所述，我们推测咖啡因增强的活性氧上调了透镜和眼周组织中的Sema 3A-Nrp 1表达，导致角膜细胞凋亡，并伴随其对三叉神经感觉纤维侵入发育中的角膜的化学排斥作用。
In this study, we investigated the effect of caffeine overexposure on corneal innervation in the early chicken embryo. Caffeine administration restricted corneal innervation by affecting trigeminal nerve development. Immunohistochemistry for phospho‐Histone3 (pHIS3) and C‐caspase3 revealed that cell survival was repressed by caffeine administration. Whole‐mount in situ hybridization against semaphorin 3A (Sema3A) and neuropilin‐1 (Nrp1) showed that both caffeine and 2,2′‐azobis(2‐methylpropionamidine) dihydrochloride (AAPH, a free radical generator) administration upregulates the expression of both Sema3A and Nrp1. Next, we demonstrated that lens ablation in the developing chicken embryos significantly affected NF‐labeled periocular nerve fascicles and innervation to the central eye region. Subsequently, we used a neuroblastoma cell line to investigate in vitro whether or not Sema3A–Nrp1 signaling exerts a key role on the caffeine‐suppressed neuron survival. Knocking‐down Sema3A through transfection with Sema3A‐siRNA dramatically decreased the responsiveness of cells to caffeine administration, as well as cell apoptosis. We suggest that Sema3A–Nrp1 signaling regulates Trp53 and Cdkn1a through Slit2–Robo1 and Ephb2. Taken together, we speculate here that caffeine‐enhanced reactive oxygen species upregulates Sema3A–Nrp1 expression in the lens and periocular tissues, resulting in corneal cell apoptosis, accompanied by its chemorepellent role on the invasion of the developing cornea by trigeminal sensory fibers.