过刊浏览

溶血磷脂酸促进离体大鼠胚胎神经干细胞的增殖及其向胆碱能神经元分化

崔慧林, 乔健天

山西医科大学神经生物学研究室.山西,太原 030001

摘要

溶血磷脂酸（lysophosphatidic acid，LPA）是一种细胞外磷脂信号。该研究用[~(3)H]--胸腺嘧啶掺入法、免疫细胞化学和Western blot等技术，观察了LPA对体外培养的大鼠胚胎神经干细胞（neural stem cells，NSCs）的增殖以及向MAF2标记的一般神经元和ChAT标记的胆碱能神经元的分化的影响。结果显示：（1）在特殊的无血清培养基中加入低浓度的LPA（0.01---1.0#mu#mol/L）后，NSCs对[~(3)H]--胸腺嘧啶的摄入呈剂量依赖性增加，表明LPA对NSCs有显著的促增殖作用；（2）在培养基中加入胎牛血清以诱导NSCs的分化，发现低浓度的LPA增加MAF2阳性和ChAT阳性神经元的比例，0.1#mu#mol／L LPA引起的增加达到峰值；（3）Western blot分析显示LPA促进了MAP2和ChAT的表达；（4）在诱导NSCs出现分化早期，用倒置显微镜观察到低浓度的LPA明显促进细胞突起的生长和细胞的迁移。以上结果表明，低浓度LPA在一定范围内可以促进NSCs的增殖、并分化为一般的MAP2阳性神经元和特殊的胆碱能神经元，而且LPA可以促进在分化早期出现的神经元或神经胶质细胞前体细胞的迁移和突起生长。

关键词： 分化; 增殖; 胆碱能神经元; 胚胎神经干细胞; 溶血磷脂酸

Promotive action of lysophosphatidic acid on proliferation of rat embryonic neural stem cells and their differentiation to cholinergic neurons {sl in vitro}

Cui Huilin, Qiao Jiantian

Department of Neurobiology, Shanxi Medical University.Taiyuan 030001,Shanxi

Abstract

Effects of lysophosphatidic acid （LPA）, an extracellular phospholipid signal, on the proliferation of rat embryonic neural stem cells （NSCs） and their differentiation into microtubule-associated protein 2 （MAP2）-positive and choline acetyltransferase （ChAT）-positive, i.e. cholinergic-committed neurons, were observed in vitro by [~(3)H]-thymidine incorporation, immunocytochemistry, Western blot and other techniques. The results showed that： （1） Lower concentrations of LPA （0.01--1.0 #mu#mol/L） dose-dependently enhanced the uptake of [~(3)H]-thymidine by NSCs cultured in specific serum-free medium, indicating a significant promotive action of LPA on the proliferation of NSCs. （2） After fetal bovine serum which induces and commences the differentiation of NSCs, was used in the medium, the lower concentrations of LPA increased the percentages of both MAP2- and ChAT-immunoreactive neurons, with a peak at 0.1 #mu#mol/L LPA in two cases. （3） The promotive effects of LPA on the differentiation of MAP2- and ChAT-positive neurons were also supported by the up-regulation of the expressions of both MAP2 and ChAT proteins detected by Western blot. （4） At the early phase of differentiation of NSCs, the cell migration and neurite extension were enhanced significantly by lower dosages of LPA under phase-contrast microscope. These results suggest that LPA within certain lower range of concentrations promotes the proliferation of NSCs and their differentiation into unspecific MAP2-positive and specific cholinergic-committed neurons, and also strengthens the migration and neurite extension of the newly-generated neuronal （and also glial as reported elsewhere） progenitors.

Key words: Differentiation;Proliferation;Cholinergic neuron;embryonic neural stem cells;lysophosphatidic acid

收稿日期：　　录用日期：

通讯作者：　　E-mail:

引用本文：

崔慧林, 乔健天. 溶血磷脂酸促进离体大鼠胚胎神经干细胞的增殖及其向胆碱能神经元分化[J]. 生理学报 2006; 58 (6): .

Cui Huilin, Qiao Jiantian. Promotive action of lysophosphatidic acid on proliferation of rat embryonic neural stem cells and their differentiation to cholinergic neurons {sl in vitro}. Acta Physiol Sin 2006; 58 (6): (in Chinese with English abstract).