Department of Neuroscience, Uppsala University, Uppsala, Sweden.
PLoS One. 2012;7(1):e29771. doi: 10.1371/journal.pone.0029771. Epub 2012 Jan 3.
Identifying external factors that can be used to control neural stem cells division and their differentiation to neurons, astrocytes and oligodendrocytes is of high scientific and clinical interest. Here we show that the Nogo-66 receptor interacting protein LINGO-1 is a potent regulator of neural stem cell maturation to neurons. LINGO-1 is expressed by cortical neural stem cells from E14 mouse embryos and inhibition of LINGO-1 during the first days of neural stem cell differentiation results in decreased neuronal maturation. Compared to neurons in control cultures, which after 6 days of differentiation have long extending neurites, neurons in cultures treated with anti-LINGO-1 antibodies retain an immature, round phenotype with only very short processes. Furthermore, neutralization of LINGO-1 results in a threefold increase in βIII tubulin-positive cells compared to untreated control cultures. By using BrdU incorporation assays we show that the immature neurons in LINGO-1 neutralized cultures are dividing neuroblasts. In contrast to control cultures, in which no cells were double positive for βIII tubulin and BrdU, 36% of the neurons in cultures treated with anti-LINGO-1 antibodies were proliferating after three days of differentiation. TUNEL assays revealed that the amount of cells going through apoptosis during the early phase of differentiation was significantly decreased in cultures treated with anti-LINGO-1 antibodies compared to untreated control cultures. Taken together, our results demonstrate a novel role for LINGO-1 in neural stem cell differentiation to neurons and suggest a possibility to use LINGO-1 inhibitors to compensate for neuronal cell loss in the injured brain.
确定能够控制神经干细胞分裂及其分化为神经元、星形胶质细胞和少突胶质细胞的外部因素具有重要的科学和临床意义。在这里，我们表明 Nogo-66 受体相互作用蛋白 LINGO-1 是神经干细胞向神经元成熟的有效调节剂。LINGO-1 由来自 E14 天小鼠胚胎的皮质神经干细胞表达，并且在神经干细胞分化的最初几天抑制 LINGO-1 会导致神经元成熟减少。与对照组培养物中的神经元相比，在分化 6 天后，具有长延伸突起的神经元在具有抗 LINGO-1 抗体的培养物中保留不成熟的圆形表型，仅有非常短的突起。此外，与未经处理的对照培养物相比，中和 LINGO-1 导致βIII 微管蛋白阳性细胞增加三倍。通过使用 BrdU 掺入测定法，我们表明 LINGO-1 中和培养物中的不成熟神经元是正在分裂的神经母细胞。与对照培养物不同，在其中没有细胞同时对βIII 微管蛋白和 BrdU 呈阳性，在分化三天后，用抗 LINGO-1 抗体处理的培养物中 36%的神经元正在增殖。TUNEL 测定显示，在用抗 LINGO-1 抗体处理的培养物中，在分化的早期阶段通过细胞凋亡的细胞数量与未经处理的对照培养物相比显著减少。总之，我们的结果表明 LINGO-1 在神经干细胞向神经元分化中的新作用，并表明可以使用 LINGO-1 抑制剂来补偿受损大脑中的神经元细胞丢失。
