Neuronal cells derived from human induced pluripotent stem cells as a functional tool of melanocortin system. (October 2017)

• Record Type: Journal Article
• Authors: Yamada-Goto, Nobuko; Ochi, Yukari; Katsuura, Goro; Yamashita, Yui; Ebihara, Ken; Noguchi, Michio; Fujikura, Junji; Taura, Daisuke; Sone, Masakatsu; Hosoda, Kiminori; Gottschall, Paul E.; Nakao, Kazuwa
• Is Part Of: Neuropeptides. Volume 65(2017)
• Page Start: 10
• Page End: 20
• Publication Date: 2017-10
• Subjects: Human iPS cells -- Neuronal cells -- Neuropeptides -- Melanocortin system -- cAMP -- Food intake; Neuropeptides -- Periodicals; Neuropeptides; Neuropeptides -- Périodiques; Neuropeptides; Electronic journals; Periodicals; 572.65
• DOI: 10.1016/j.npep.2017.04.004
• Languages: English
• ISSNs: 0143-4179

Abstract

Abstract: Background: The preparation of human neurons derived from human induced pluripotent stem (iPS) cells can serve as a potential tool for evaluating the physiological and pathophysiological properties of human neurons and for drug development. Methods: In the present study, the functional activity in neuronal cells differentiated from human iPS cells was observed. Results: The differentiated cells expressed mRNAs for classical neuronal markers (microtubule-associated protein 2, β-tubulin III, calbindin 1, synaptophysin and postsynaptic density protein 95) and for subunits of various excitatory and inhibitory transmitters (NR1, NR2A, NR2B, GABAA α1). Moreover, the differentiated cells expressed neuropeptides and receptors which are predominantly present in the hypothalamus. The expression of mRNA for preopiomelanocortin, agouti-related protein (AgRP), melanocortin-3 receptor (MC3R) and melanocortin-4 receptor (MC4R) increased in culture with a peak on Day 30 which subsequently decreased at Day 45. Immunoreactivities for MC3R and MC4R were also observed in cells differentiated from human iPS cells. Application of a potent agonist for MC3R and MC4R, [Nle4, D-Phe7]-α-melanocyte-stimulating hormone, significantly increased intracellular cAMP levels, but this was suppressed by AgRP (83-132) and SHU9119. Conclusions: These findings offer the possibility for drug developments using neurons differentiated from normal or disease-associated human iPS cells. Highlights: The functional activity of melanocortin system in neuronal cells differentiated from human iPS cells was observed. After application of a differentiation medium containing neurotrophic factors, we observed mRNA expression for various neuronal markers including, MAP2, β-tubulin III, calbindin 1, synaptophysin and PSD-95, but not glial fibrillary acid as an astrocyte marker. The neuronal cells expressed mRNA of POMC, AgRP, MC3R and MC4R, which were increased during cultures with a peak on Day 30 and subsequently decreased to Day 45. Application of a potent agonist for MC3R and MC4R, [Nle4, D-Phe7]-α-melanocyte-stimulating hormone, significantly increased intracellular cAMP levels, but this was suppressed by AgRP (83-132) and SHU9119, which are, respectively, an inverse agonist and an antagonist for MC3R and MC4R.