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S100A9促进胚胎干细胞向神经前体细胞分化中顶端-基底极化和神经管形成
S100A9 promotes apical-basal polarity and neural tube formation during differentiation of embryonic stem cells into neural progenitor cells
- 陆军军医大学学报 2023年45卷第1期 页码：10-18
- 作者机构：
400010 重庆，重庆医科大学附属第二医院神经外科
- 作者简介：
[黄宁, E-mail: 304237@hospital.cqmu.edu.cnHUANG Ning, E-mail: 304237@hospital.cqmu.edu.cn
- 基金信息：重庆市自然科学基金面上项目(cstc2021jcyj-msxmX0036)
DOI：10.16016/j.2097-0927.202204002
中图分类号： R322.8;R318;R329.24收稿：2022-04-01，
修回：2022-06-08，
纸质出版：2023-01-15
- 稿件说明：
移动端阅览
张翔, 岳建和, 黄宁. S100A9促进胚胎干细胞向神经前体细胞分化中顶端-基底极化和神经管形成[J]. 陆军军医大学学报, 2023,45(1):10-18.
Xiang ZHANG, Jianhe YUE, Ning HUANG. S100A9 promotes apical-basal polarity and neural tube formation during differentiation of embryonic stem cells into neural progenitor cells[J]. Journal of Army Medical University, 2023, 45(1): 10-18.
张翔, 岳建和, 黄宁. S100A9促进胚胎干细胞向神经前体细胞分化中顶端-基底极化和神经管形成[J]. 陆军军医大学学报, 2023,45(1):10-18. DOI： 10.16016/j.2097-0927.202204002.
Xiang ZHANG, Jianhe YUE, Ning HUANG. S100A9 promotes apical-basal polarity and neural tube formation during differentiation of embryonic stem cells into neural progenitor cells[J]. Journal of Army Medical University, 2023, 45(1): 10-18. DOI： 10.16016/j.2097-0927.202204002.
摘要
目的
2
探讨胚胎干细胞(embryonic stem cells
ESCs)向神经前体细胞(neural progenitor cells
NPCs)分化过程中S100A9促进顶端-基底极化(apical-basal polarity
ABP)和神经管结构形成的作用。
方法
2
采用免疫荧光观察体外培养ESCs向NPCs分化1、2、3和6 d时细胞管腔样结构排列，神经巢蛋白(Nestin)和配对盒因子6(paired box protein 6
Pax-6)表达，以及S100A9、N-钙粘蛋白(N-cadherin)和zonula occludens 1(ZO-1)定位变化。慢病毒转染下调S100A9及外源性S100A9重组蛋白干预细胞后检测对N-cadherin和ZO-1定位的影响。Western blot分析S100A9在ESCs向NPCs分化过程中不同时间点表达差异，同时观察S100A9对Notch1及下游centromere binding factor 1(CBF1)的调控。
结果
2
ESCs在体外定向分化后第3天起表达Nestin和Pax-6，N-cadherin和ZO-1定位于顶端侧并呈现出ABP特征性神经玫瑰花状结构(neural rosette)，细胞在第6天形成神经管结构。S100A9在分化后第3天起表达明显增加(
P
<
0.01)并定位于顶端侧。下调S100A9后ESCs仍能分化为NPCs，但neural rosette数量明显减少(
P
<
0.01)。予以外源性S100A9重组蛋白可部分恢复neural rosette数量(
P
<
0.01)。在此分化过程中，下调S100A9降低Notch1及下游CBF1表达。
结论
2
S100A9可能通过调控Notch1/CBF1参与ESCs向NPCs分化过程中ABP和神经管结构形成。
Abstract
Objective
2
To explore the role of S100A9 in the apical-basal polarity (ABP) and neural tube formation during the differentiation of embryonic stem cells (ESCs) into neural progenitor cells (NPCs)
in vitro
.
Methods
2
Immunofluorescence assay was used to observe the luminal arrangement
expression of Nestin and paired box protein 6 (Pax-6)
and localization of S100A9
N-cadherin and Zonula occludens 1 (ZO-1) on days 1
2
3 and 6 of the ESCs differentiation into NPCs. In addition
the ESCs were treated with S100A9-downregulation lentivirus as well as subsequential exogenous S100A9 recombinant protein
then the localization changes of N-cadherin and ZO-1 were detected. The S100A9 expression in different time points during ESCs differentiation process was measured by Western blotting
and the effects of S100A9 on Notch1 and downstream centromere binding factor 1 (CBF1) were also tested.
Results
2
Nestin and Pax-6 were positively expressed after ESCs differentiation for 3 d
N-cadherin and ZO-1 were gathered at the apical side and presented the characteristic neural rosette structure of ABP
and the neural tubes were formed on day 6. The expression of S100A9 was upregulated significantly from day 3 after differentiation (
P
<
0.01) and was located on the apical side. Although ESCs were still able to differentiate into NPCs after down-regulation of S100A9
the number of neural rosettes was greatly reduced (
P
<
0.01)
which could be partially restored by supplement of exogenous S100A9 recombinant protein (
P
<
0.01). Furthermore
declined S100A9 inhibited the expression of Notch 1 and downstream CBF1 during differentiation.
Conclusion
2
S100A9 may contribute to the ABP and the formation of neural lumen during the differentiation of ESCs into NPCs by regulating Notch 1/CBF1.
关键词
Keywords
references
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