The Molecular Mechanisms Underlying Prostaglandin D2-Induced Neuritogenesis in Motor Neuron-Like NSC-34 Cells

Abstract

Prostaglandins are a group of physiologically active lipid compounds derived from arachidonic acid. Our previous study has found that prostaglandin E2 promotes neurite outgrowth in NSC-34 cells, which are a model for motor neuron development. However, the effects of other prostaglandins on neuronal differentiation are poorly understood. The present study investigated the effect of prostaglandin D2 (PGD2) on neuritogenesis in NSC-34 cells. Exposure to PGD2 resulted in increased percentages of neurite-bearing cells and neurite length. Although D-prostanoid receptor (DP) 1 and DP2 were dominantly expressed in the cells, BW245C (a DP1 agonist) and 15(R)-15-methyl PGD2 (a DP2 agonist) had no effect on neurite outgrowth. Enzyme-linked immunosorbent assay demonstrated that PGD2 was converted to 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) under cell-free conditions. Exogenously applied 15d-PGJ2 mimicked the effect of PGD2 on neurite outgrowth. GW9662, a peroxisome proliferator-activated receptor–gamma (PPARγ) antagonist, suppressed PGD2-induced neurite outgrowth. Moreover, PGD2 and 15d-PGJ2 increased the protein expression of Islet-1 (the earliest marker of developing motor neurons), and these increases were suppressed by co-treatment with GW9662. These results suggest that PGD2 induces neuritogenesis in NSC-34 cells and that PGD2-induced neurite outgrowth was mediated by the activation of PPARγ through the metabolite 15d-PGJ2.

An in vivo drug screen reveals that cyclooxygenase 2-derived prostaglandin D2promotes spinal cord neurogenesis

“…Also, in vitro work using the mouse neuroblastoma and spinal cord motor neuron fusion cell line (NSC-34), has shown that PGE 2 increases the percentage of neurite-bearing cells (Nango et al, 2017) and the differentiation of NSC-34 cells into functional motor neurons (Nango et al, 2020a). In the same cell line, PGD 2 also increased the proportion of cells with neurites and neurite length in NSC-34 cell cultures (Nango et al 2020b). Therefore, a possible role for PGD 2 signalling in promoting neuronal differentiation in the developing spinal cord should not be excluded.…”

CHI3L1 signaling impairs hippocampal neurogenesis and cognitive function in autoimmune-mediated neuroinflammation

“…In vitro, we used the shRNA against CRTH2 and the selective CRTH2 antagonist AZD1981 to firmly establish the ligand-receptor relationship for CHI3L1-CRTH2. In vivo, both approaches intended to block CRTH2 activation by CHI3L1 binding also potentially obstructed the prostaglandin D2 signaling, which is involved in a wide variety of neurophysiological functions ( 77 ), and, in the unusual context of neuronal differentiation, was counterintuitively described to be able to promote outgrowth of neuronal processes in immortalized neuronal cells via an unidentified receptor ( 78 ). While we could not verify such antagonizing effect of prostaglandin D2 in our NSCs, our findings on CRTH2 blocking solidly suggested that, at least in an inflammatory condition, CRTH2 is predominantly activated by CHI3L1 to mediate the inhibition of neurogenesis.…”

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

“…A neurite outgrowth analysis was performed according to our previous method [ 20 , 22 ]. Briefly, phase-contrast micrographs were captured by inverted microscopy (IX71, Olympus, Tokyo, Japan), after which 50 cells per condition were randomly chosen for counting neurite-bearing cells.…”

“…The Western blot analysis was performed as described previously [ 20 , 22 ]. Briefly, protein extracts from the cells treated with vehicle (EtOH) for 2 days, 30 μM PGE 2 for 2 days, or 10 μM RA for 7 days were separated on SDS-polyacrylamide gels and transferred to polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA) using a Transblot SD Semi-Dry Transfer Cell (BioRad, Hercules, CA, USA).…”