Prenatal ethanol exposure can result in loss of neural stem cells (NSCs) and decreased brain growth. Here, we assessed whether a noncoding RNA (ncRNA) related to the NSC self-renewal factor Oct4/Pou5f1, and transcribed from a processed pseudogene locus on mouse chromosome 9 (mOct4pg9), contributed to the loss of NSCs due to ethanol. Mouse fetal cortical-derived NSCs, cultured ex vivo to mimic the early neurogenic environment of the fetal telencephalon, expressed mOct4pg9 ncRNA at significantly higher levels than the parent Oct4/Pou5f1 mRNA. Ethanol exposure increased expression of mOct4pg9 ncRNA, but decreased expression of Oct4/Pou5f1. Gain- and loss-of-function analyses indicated that mOct4pg9 overexpression generally mimicked effects of ethanol exposure, resulting in increased proliferation and expression of transcripts associated with neural maturation. Moreover, mOct4pg9 associated with Ago2 and with miRNAs, including the anti-proliferative miR-328-3p, whose levels were reduced following mOct4pg9 overexpression. Finally, mOct4pg9 inhibited Oct4/Pou5f1–3’UTR-dependent protein translation. Consistent with these observations, data from single-cell transcriptome analysis showed that mOct4pg9-expressing progenitors lack Oct4/Pou5f1, but instead overexpress transcripts for increased mitosis, suggesting initiation of transit amplification. Collectively, these data suggest that the inhibitory effects of ethanol on brain development are explained, in part, by a novel ncRNA which promotes loss of NSC identity and maturation.

产前接触乙醇会导致神经干细胞 (NSC) 损失和大脑生长下降。在这里，我们评估了与 NSC 自我更新因子 Oct4/Pou5f1 相关的非编码 RNA (ncRNA)，并从小鼠 9 号染色体上经过加工的假基因位点 ( mOct4pg9 ) 转录，是否导致乙醇导致 NSC 损失。离体培养的小鼠胎儿皮质源性 NSC 模拟胎儿端脑的早期神经源性环境，其m Oct4pg9 ncRNA 的表达水平显着高于亲本 Oct4/Pou5f1 mRNA。乙醇暴露增加了m Oct4pg9 ncRNA 的表达，但降低了 Oct4/Pou5f1 的表达。功能获得和丧失分析表明， m Oct4pg9 过表达通常模仿乙醇暴露的效果，导致与神经成熟相关的转录物增殖和表达增加。此外， m Oct4pg9 与 Ago2 和 miRNA（包括抗增殖 miR-328-3p）相关，其水平在m Oct4pg9 过表达后降低。最后， m Oct4pg9 抑制 Oct4/Pou5f1–3'UTR 依赖性蛋白翻译。与这些观察结果一致，来自单细胞转录组分析的数据表明，表达m Oct4pg9 的祖细胞缺乏 Oct4/Pou5f1，而是过度表达转录物以增加有丝分裂，表明转运扩增的启动。总的来说，这些数据表明，乙醇对大脑发育的抑制作用部分可以通过一种新型 ncRNA 来解释，这种新型 ncRNA 会促进 NSC 身份的丧失和成熟。