A Novel Role of Nuclear and Membrane Receptor on Isoflavone-Induced Neuritogenesis and Synaptogenesis
A Novel Role of Nuclear and Membrane Receptor on Isoflavone-Induced Neuritogenesis and Synaptogenesis
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DOI：
10.1210/jendso/bvab048.1632
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发表时间：
2021-05-03
影响因子：
4.1
通讯作者：
Koibuchi N
中科院分区：
文献类型：
--
作者：
Ariyani W;Miyazaki W;Koibuchi N
甲状腺激素受体（TR）和雌激素受体（ER）在脑发育中起着重要作用。TR和ER参与神经元中树突生长、棘和突触形成。大豆异黄酮，如染料木素、大豆苷元和大豆苷元代谢物S-雌马酚，已知通过TR、ER和GPER 1（一种G蛋白偶联的ER）发挥其作用。然而，对脑发育，特别是在神经突和突触发生过程中的作用机制，还没有得到广泛的研究。我们使用小鼠原代小脑培养物、星形胶质细胞富集培养物、Neuro-2A克隆细胞以及与神经元和星形胶质细胞共培养物来评估马槟榔酮的作用。大豆异黄酮增强TH或雌二醇（E2）介导的浦肯野细胞树突分枝。在两种培养物中，G15（一种选择性GPER 1拮抗剂）和ICI 182.780（一种ER拮抗剂）抑制了这种增强。核TRs或ERs的敲低也显著减少浦肯野细胞的树突分枝。它还增加了TH反应基因的mRNA水平，包括Mbp，Bdnf，Rc 3，Ntf 3，Camk 2b，Hr，以及参与突触可塑性的Syn 1，Syp和Psd 95。异黄酮还增加了突触蛋白-1，突触素，和PSD 95在小脑培养的树突和膜部分的蛋白水平。为了进一步研究分子机制，我们使用Neuro-2A克隆细胞。异黄酮还诱导Neuro-2A的神经突生长。通过RNAi敲低TRs、ERs和GPR 30减少了孕酮诱导的神经突生长。此外，Neuro-2A和星形胶质细胞的共培养研究也显示了孕酮诱导的神经突生长的增加。此外，在Neuro-2A分化过程中，β-胡萝卜酮增加了突触蛋白-1或突触素和F-肌动蛋白在丝状伪足尖端的定位。核ER或GPR 30的敲低显著降低了轴突和膜组分中的丝状伪足和突触蛋白-1或突触素表达水平的数量。然而，与星形胶质细胞共培养后对丝状伪足形成没有显著影响。这些结果表明，核ER和TR在孕酮诱导的轴突发生中起着重要作用。通过膜受体和F-actin的非基因组信号传导是突触发生所必需的。星形胶质细胞-神经元通讯也增加了多巴胺诱导的轴突发生，但不增加突触发生。
Thyroid hormone (TH) receptor (TR) and estrogen receptor (ER) play crucial roles in brain development. TR and ER are involved in dendrite growth, spines, and synapse formation in neurons. Soybean isoflavones, such as genistein, daidzein, and daidzein metabolite, S-equol are known to exert their action through TR, ER, and GPER1, a G-protein-coupled ER. However, the mechanisms of isoflavones action on brain development, especially during neuritogenesis and synaptogenesis, have not yet been extensively studied. We evaluated the effects of isoflavones using mouse primary cerebellar culture, astrocyte-enriched culture, Neuro-2A clonal cells, and co-culture with neurons and astrocytes. Soybean isoflavone augmented TH- or estradiol (E2)-mediated dendrite arborization of Purkinje cells. Such augmentation was suppressed by G15, a selective GPER1 antagonist, and ICI 182.780, an antagonist for ERs in both cultures. The knockdown of nuclear TRs or ERs also significantly reduced the dendrite arborization of Purkinje cells. It also increased the mRNA levels of TH-responsive genes, including Mbp, Bdnf, Rc3, Ntf3, Camk2b, Hr, and also Syn1, Syp, and Psd95 that are involved in synaptic plasticity. Isoflavones also increased the protein levels of synapsin-1, synaptophysin, and PSD95 in dendrite and membrane fraction of the cerebellar culture. To study further the molecular mechanism, we used Neuro-2A clonal cells. Isoflavones also induced neurite growth of Neuro-2A. The knockdown of TRs, ERs, and GPR30 by RNAi reduced isoflavones-induced neurite growth. Moreover, the co-culture study of Neuro-2A and astrocytes also showed an increase in isoflavones-induces neurite growth. In addition, isoflavones increased the localization of synapsin-1 or synaptophysin and F-actin in filopodia tips during Neuro-2A differentiation. The knockdown of nuclear ERs or GPR30 significantly reduced the number of filopodia and synapsin-1 or synaptophysin expression levels in neurite and membrane fractions. However, there are no significant effects of filopodia formation after co-culture with astrocytes. These results indicate that nuclear ERs and TRs play an essential role in isoflavones-induces neuritogenesis. Non-genomics signaling through membrane receptor and F-actin are necessary for the isoflavones-induces synaptogenesis. Astrocytes-neurons communication also increased isoflavones-induced neuritogenesis, but not synaptogenesis.