Notch1 对全反式视黄酸诱导的神经管缺陷和神经干细胞分化的影响。

Effect of Notch1 on neural tube defects and neural stem cell differentiation induced by all‑<em>trans</em> retinoic acid.

机构信息

Department of Pediatrics, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong 250021, P.R. China.

Department of Pediatrics, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250021, P.R. China.

出版信息

Mol Med Rep. 2021 Mar;23(3). doi: 10.3892/mmr.2021.11859. Epub 2021 Jan 21.

DOI:10.3892/mmr.2021.11859

PMID:33655333

Abstract

Neural tube defects (NTDs) are the most serious and common birth defects in the clinical setting. The Notch signaling pathway has been implicated in different processes of the embryonic neural stem cells (NSCs) during neural tube development. The aim of the present study was to investigate the expression pattern and function of Notch1 (N1) in all‑trans retinoic acid (atRA)‑induced NTDs and NSC differentiation. A mouse model of brain abnormality was established by administering 28 mg/kg atRA, and then brain development was examined using hematoxylin and eosin (H&E) staining. The N1 expression pattern was detected in the brain of mice embryos via immunohistochemistry and western blotting. NSCs were extracted from the fetal brain of C57 BL/6 embryos at 18.5 days of pregnancy. N1, Nestin, neurofilament (NF), glial fibrillary acidic protein (GFAP) and galactocerebroside (GALC) were identified using immunohistochemistry. Moreover, N1, presenilin 1 (PS1), Nestin, NF, GFAP and GALC were detected via western blotting at different time points in the NSCs with control media or atRA media. H&E staining identified that the embryonic brain treated with atRA was more developed compared with the control group. N1 was downregulated in the embryonic mouse brain between days 11 and 17 in the atRA‑treated group compared with the untreated group. The distribution of N1, Nestin, NF, GFAP and GALC was positively detected using immunofluorescence staining. Western blotting results demonstrated that there were significantly, synchronous decreased expression levels of N1 and PS1, but increased expression levels of NF, GFAP and GALC in NSCs treated with atRA compared with those observed in the controls (P<0.05). The results suggested that the N1 signaling pathway inhibited brain development and NSC differentiation. Collectively, it was found that atRA promoted mouse embryo brain development and the differentiation of NSCs by inhibiting the N1 pathway.

摘要

神经管缺陷（NTDs）是临床中最严重和常见的出生缺陷。Notch 信号通路已被牵涉到胚胎神经干细胞（NSCs）在神经管发育过程中的不同过程。本研究的目的是研究 Notch1（N1）在全反式视黄酸（atRA）诱导的 NTDs 和 NSC 分化中的表达模式和功能。通过给予 28mg/kg atRA 建立脑异常的小鼠模型，然后通过苏木精和伊红（H&E）染色检查脑发育。通过免疫组织化学和蛋白质印迹法检测 N1 在小鼠胚胎脑中的表达模式。从 C57BL/6 胚胎 18.5 天的胎脑中提取 NSCs。通过免疫组织化学鉴定 N1、巢蛋白（Nestin）、神经丝（NF）、胶质纤维酸性蛋白（GFAP）和半乳糖脑苷脂（GALC）。此外，在对照培养基或 atRA 培养基中的 NSCs 中，在不同时间点通过 western blot 检测 N1、早老素 1（PS1）、Nestin、NF、GFAP 和 GALC。H&E 染色发现用 atRA 处理的胚胎脑比对照组发育得更好。与未处理组相比，atRA 处理组的胚胎鼠脑中 N1 在第 11 天至第 17 天之间下调。免疫荧光染色阳性检测 N1、巢蛋白、NF、GFAP 和半乳糖脑苷脂的分布。Western blot 结果表明，与对照组相比，atRA 处理的 NSCs 中 N1 和 PS1 的表达水平显著且同步下降，而 NF、GFAP 和 GALC 的表达水平升高（P<0.05）。结果表明，N1 信号通路抑制了脑发育和 NSC 分化。总之，研究发现 atRA 通过抑制 N1 通路促进了小鼠胚胎脑发育和 NSCs 的分化。