Neutralization of LINGO-1 during In Vitro Differentiation of Neural Stem Cells Results in Proliferation of Immature Neurons

Camilla Lööv,Niklas Marklund,Maria Fernqvist,Adrian Walmsley,Anna Erlandsson,Alain Chédotal

doi:10.1371/journal.pone.0029771

Camilla Lööv, Niklas Marklund + Show 4 more

Open Access

https://doi.org/10.1371/journal.pone.0029771

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Journal: PloS one	Publication Date: Jan 3, 2012
Citations: 69	License type: CC BY 4.0

Affiliation: Uppsala University, Novartis (Switzerland)

Abstract

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.

Highlights

Several important breakthroughs during recent years have raised a hope that stem cell-based therapies could be used to restore function and integrity after acute brain injury and other disorders of the central nervous system
Cell lysates were prepared from neural stem and progenitor cells (NSPCs) proliferating in the presence of the mitogens epidermal growth factor (EGF) and FGF2 and from NSPCs that have differentiated in the absence of the mitogens for 1, 3, 6 and 9 days
We report a novel function for LINGO-1 in neural stem cell differentiation, regulating the maturation of progenitor cells differentiating along the neuronal lineage

Summary

Introduction

Several important breakthroughs during recent years have raised a hope that stem cell-based therapies could be used to restore function and integrity after acute brain injury and other disorders of the central nervous system. In order to develop effective and safe regenerative treatments it is necessary to identify factors that could be used to control differentiation, proliferation and survival of neural stem and progenitor cells (NSPCs). Activation of the Wnt pathway has been demonstrated to direct neural cortical progenitor cells to differentiate to neurons in vitro and to promote hippocampal neurogenesis in vivo but the Wnt ligands has been shown to induce proliferation of neural stem cells [9,10,11,12,13,14]. Platelet derived growth factor (PDGF) was earlier suggested to be involved in neuronal differentiation, but has more recently been shown to rather promote proliferation of precursor cells [15,16,17]

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