Highly Efficient Conversion of Motor Neuron-Like NSC-34 Cells into Functional Motor Neurons by Prostaglandin E2.

Hiroshi Nango,Masaki Sato,Yuri Aono,Yasuhiro Kosuge,Yoshihisa Ito,Tadashi Saigusa,Yoshiyuki Shibukawa,Kumiko Ishige

doi:10.3390/cells9071741

Hiroshi Nango, Masaki Sato + Show 6 more

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https://doi.org/10.3390/cells9071741

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Abstract

Motor neuron diseases are a group of progressive neurological disorders that degenerate motor neurons. The neuroblastoma × spinal cord hybrid cell line NSC-34 is widely used as an experimental model in studies of motor neuron diseases. However, the differentiation efficiency of NSC-34 cells to neurons is not always sufficient. We have found that prostaglandin E2 (PGE2) induces morphological differentiation in NSC-34 cells. The present study investigated the functional properties of PGE2-differentiated NSC-34 cells. Retinoic acid (RA), a widely-used agent inducing cell differentiation, facilitated neuritogenesis, which peaked on day 7, whereas PGE2-induced neuritogenesis took only 2 days to reach the same level. Whole-cell patch-clamp recordings showed that the current threshold of PGE2-treated cell action potentials was lower than that of RA-treated cells. PGE2 and RA increased the protein expression levels of neuronal differentiation markers, microtubule-associated protein 2c and synaptophysin, and to the same extent, motor neuron-specific markers HB9 and Islet-1. On the other hand, protein levels of choline acetyltransferase and basal release of acetylcholine in PGE2-treated cells were higher than in RA-treated cells. These results suggest that PGE2 is a rapid and efficient differentiation-inducing factor for the preparation of functionally mature motor neurons from NSC-34 cells.

Highlights

Motor neurons (MNs) reside in the central nervous system and control muscle activities [1].There are two types of MNs: upper and lower MNs
We have previously reported that prostaglandin E2 (PGE2 ), a lipid mediator produced from arachidonic acid, induced cell death in differentiated NSC-34 cells [13,14]
We compared the effects of PGE2 and Retinoic acid (RA) on neurite outgrowth in undifferentiated NSC-34 cells, and evaluated the time-dependency of PGE2 - and RA-induced neurite outgrowth by phase-contract microscopy

Summary

Introduction

Motor neurons (MNs) reside in the central nervous system and control muscle activities [1]. There are two types of MNs: upper and lower MNs. In particular, lower MNs, located in the ventral horn of the spinal cord gray matter, form unique and irreplaceable function axons with irreplaceable functions that project directly to the endplate of peripheral muscles [2]. On receiving glutamate from upper MNs, lower MNs fire action potentials, releasing acetylcholine (ACh) at. Cells 2020, 9, 1741 the neuromuscular junctions. MNs isolated from mouse spinal cords are identified as long neurite-bearing cells that express neuronal markers including neuronal cytoskeletal proteins, neurofilaments, and choline acetyltransferase (ChAT), in addition to MN-specific markers such as HB9 and Islet-1 [3]

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