Modulated DISP3/PTCHD2 expression influences neural stem cell fate decisions

Jana Konířová,Jana Oltová,Justyna Kopycińska,Michal Kolář,Martina Zíková,Petr Bartůněk,Alicia Corlett

doi:10.1038/srep41597

Jana Konířová, Jana Oltová + Show 5 more

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https://doi.org/10.1038/srep41597

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Abstract

Neural stem cells (NSCs) are defined by their dual ability to self-renew through mitotic cell division or differentiate into the varied neural cell types of the CNS. DISP3/PTCHD2 is a sterol-sensing domain-containing protein, highly expressed in neural tissues, whose expression is regulated by thyroid hormone. In the present study, we used a mouse NSC line to investigate what effect DISP3 may have on the self-renewal and/or differentiation potential of the cells. We demonstrated that NSC differentiation triggered significant reduction in DISP3 expression in the resulting astrocytes, neurons and oligodendrocytes. Moreover, when DISP3 expression was disrupted, the NSC “stemness” was suppressed, leading to a larger population of cells undergoing spontaneous neuronal differentiation. Conversely, overexpression of DISP3 resulted in increased NSC proliferation. When NSCs were cultured under differentiation conditions, we observed that the lack of DISP3 augmented the number of NSCs differentiating into each of the neural cell lineages and that neuronal morphology was altered. In contrast, DISP3 overexpression resulted in impaired cell differentiation. Taken together, our findings imply that DISP3 may help dictate the NSC cell fate to either undergo self-renewal or switch to the terminal differentiation cell program.

Highlights

Neural stem cells (NSCs) are defined by their dual ability to self-renew through mitotic cell division or differentiate into the varied neural cell types of the CNS
All data represent the mean of biological replicates with error bars indicating standard deviation and the level of statistical significance (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001)
Our results show that modified expression levels of DISP3 lead to altered mRNA levels of insulin-like growth factor binding protein 7 (Igfbp7), lipoyltransferase 1 (Lipt1), diacylglycerol kinase alpha (Dgka), brain serine/threonine kinase 1 (Brsk1) and endothelial differentiation sphingolipid G-protein-coupled receptor 8 (Edg8) (Fig. 5C)

Summary

Introduction

Neural stem cells (NSCs) are defined by their dual ability to self-renew through mitotic cell division or differentiate into the varied neural cell types of the CNS. We demonstrated that NSC differentiation triggered significant reduction in DISP3 expression in the resulting astrocytes, neurons and oligodendrocytes. Neural stem cells (NSCs) are defined by their ability to self-renew through mitotic cell division and to differentiate into the various neural cell types: neurons, astrocytes and oligodendrocytes[1,2]. NSCs first undergo symmetric self-renewal to expand the stem cell pool, which is followed by asymmetric neurogenic and gliogenic cell division to generate neurons and glia, respectively[3]. NS-5 cells represent tripotent NSCs, so even after prolonged expansion, they are still capable of generating neurons, astrocytes and oligodendrocytes under particular conditions in vitro[18,19]. The Dispatched 3 gene (Disp3), known as Ptchd[2] or KIAA1337, encodes a 13-transmembrane domain-containing protein, highly expressed in neural tissue and regulated both in vivo and in vitro by thyroid www.nature.com/scientificreports/

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