The effect of R-spondin3 on proliferation of neural stem cells and Wnt/β-catenin signals in mice
10.3760/cma.j.issn.1671-8925.2018.04.004
- VernacularTitle:R-脊椎蛋白3对小鼠神经干细胞增殖及Wnt/β-catenin通路的影响
- Author:
Ruifeng WANG
1
;
Haoju ZHANG
;
Yiwu DAI
;
Ruxiang XU
Author Information
1. 山西医科大学第二临床医学院
- Keywords:
R-spondin3;
Neural stem cells;
Wnt/β3-catenin signal pathway
- From:
Chinese Journal of Neuromedicine
2018;17(4):344-348
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of R-spondin3 on the proliferation of neural stem cells (NSCs) and its mechanism in mice.Methods The mouse NSCs derived from the subventricular zone of E 14-15d CD 1 mice were confirmed by immunofluorescence assay.The NSCs after 3 passages of culture were chosen and randomly divided into 2 groups (V=1 mL).In the experimental group,0.8 μL of R-spondin3 with an initial concentration of 50 μg/mL was added (final concentration:40 ng/mL) while in the control group an equal amount of culture fluid was added.The proliferation of the cells in the 2 groups was detected by 5-Bromo-2-deoxy Uridine (BrdU) kits after the cells were treated by R-spondin3 for 6 hours.The protein expression of [β-catenin was measured by western blotting after the cells were treated by R-spondin3 for 4 and 8 hours.Results Under optical microscopy,the round and bright cells grew in culture medium and easily accumulated to become neurospheres.Immunofluorescence assay showed that over 90％ of the cells expressed Nestin and SOX2 and that some of them expressed NeuN or GFAP after induced differentiation.Brdu proliferation test showed that the proliferation rate of Brdu+/DAPI+ for the experimental group (1.56±0.03) was significantly higher than that for the control group (1.04±0.04) (P＜0.05).Western blotting showed that the expression levels of [β-catenin were increased at both 4h and 8h after treatment for the experimental group (1.09±0.10 and 1.20±0.13),significantly higher than those for the control group (0.56±0.05 and 0.83±0.04) (P＜0.05).Conclusions R-spondin3 can promote in vitro proliferation of NSCs in mice,which may be associated with activated Wnt/β-catenin signal pathways.
