Abstract
The aim of the study was to obtain a neural cell population enriched in culture with cholinergic neurons and their determined precursors. It has been established that the joint application of retinoic acid and acetylcholine appears to be the most efficient combination of neuroinductors, stimulating the differentiation of cholinergic neurons from neural stem cells. During the entire period of culturing, the amount of ChAT+-positive cells increased reliably to 21.1 ± 6.2% from 5.3 ± 2.9, whereas the ir concentration in the control samples was 9.1 ± 4.8% of the total cell count. The cell population enriched with cholinergic neurons and their determined precursors correlated well with the increased acetylcholinesterase activity. Thus, the addition of retinoic acid and acetylcholine stimulate both neurogenesis and cholinogenesis in human fetal neural stem cell cultures.
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References
Perry, E.K., Jonson, M., Ekonomou, A., et al., Neurogenic abnormalities in Alzheimer’s disease differ between stages of neurogenesis and partly related to cholinergic pathology, Neurobiol. Dis, 2012, vol. 47, no. 2, pp. 155–162.
Lauder, J.M. and Schambra, U.B., Morphogenic roles of acetylcholine, Environ. Health Persp., 1999, vol. 107, pp. 65–69.
Schammbra, U.B., Sulik, K.K., Petrusz, P., and Lauder, J.M., Ontogeny of cholinergic neurons in the mouse fore-brain, J. Comp. Neurol., 1989, vol. 288, pp. 101–140.
Barald, K.F. and Berg, D.K., Hight affinity uptake by spinal cord neurons in dissociated cell cultures, Dev. Biol., 1978, vol. 65, pp. 90–99.
Honegger, P. and Richelson, E., Neurotransmitter synthesis, storage and release by aggregating cell cultures of rat brain, J. Brain Res., 1979, vol. 162, pp. 89–101.
Coleman, B.A. and Palmer, T., Regulation of acetyl-cholinesterase expression during neuronal differentiation, J. Biol. Chem., 1996, vol. 271, no. 8, pp. 4410–4416.
El Omri, A., Han, J., Kawada, K., et al., Luteolin enhances cholinergic activities in PC12 cells through ERK1/2 and PI3K/Akt pathways, J. Brain Res., 2012, vol. 1437, no. 9, pp. 16–25.
Janiesch, P.C., Kruger, H.S., Poshel, B., et al., Cholinergic control in developing prefrontal-hippocampal networks, J. Neurosci., 2011, vol. 31, no. 49, pp. 17955–17970.
Janacait, G.M., Pratt, L., Acevedo, C., and Ni, L., Microglial regulation of cholinergic differentiation in the basal forebrain, Dev. Neurobiol., 2012, vol. 72, no. 6, pp. 857–864.
Allard, S., Leon, W.C., Pakavathkumar, P., et al., Impact of the NGF maturation and degradation pathway on the cortical cholinergic system phenotype, J. Neurosci., 2012, vol. 8, no. 6, pp. 2002–2012.
Kotasova, H., Vesela, I., Kucera, J., et al., Posphoinositide-3-kinase inhibition enables retinoic-acid-induced neurogenesis in monolayer culture of embryonic stem cells, J. Cell Biochem., 2012, vol. 113, no. 2, pp. 563–570.
Bourdeaut, F., Janoueix-Lerosey, I., Lucchesi, C., et al., Cholinergic switch associated with morphological differentiation in neuroblastoma, J. Pathol., 2009, no. 4, pp. 463–472.
Bozhkova, V.P., Brezhestovskii, L.A., Buravlev, V.M., et al., Rukovodstvo po kul’tivirovaniyu nervnoi tkani (Culturing Nervous Tissue: A Guide), Moscow: Nauka, 1988.
Svendsen, C.N., Borg, M.G., Armstrong, R.E., et al., A new method for the rapid and long-term growth of human neural precursor cells, J. Neurosci. Meth., 1998, vol. 85, pp. 141–152.
Minguell, J.J., Fierro, F.A., Epunan, M.J., et al., Nonstimulated human uncommitted mesenchymal stem cells express cell markers of mesenchymal and neuronal lineages, Stem Cells Dev., 2005, vol. 14, no. 4, pp. 408–414.
Liu, X., Zhu, Y., and Gao, W., Isolation of neural stem cells from the spinal cords of low temperature preserved abortus, J. Neurosci. Meth., 2006, vol. 157, no. 1, pp. 64–70.
Ellman, G.L., Courtney, K.D., Andres, V., et al., A new and rapid colorimetric determination of acetylcholinesterase activity, Pharmacology, 1961, vol. 7, pp. 88–95.
Tsimbalyuk, V.I., Vasil’eva, I.G., Oleksenko, N.P., Chopik, N.G., Tsyubko, O.I., and Galanta, O.S., Study of differentiation potential of acetylcholinergic neurons in the stem cell culture of human embryonic brain, Ukr. Nevrol. Zh., 2010, no. 2, pp. 94–97.
Carpenter, M.K., Cui, X., Hu, Z., et al., In vitro expansion of a multipotent population of human neural precursor cells, Exp. Neurol., 1999, vol. 158, pp. 265–278.
Asai, D. and Remolana, N., Tubulin isotype usage in vivo: a unique spatial distribution of the minor neural-specific p-tubulin isotype in pheocromocytoma cells, Dev. Biol., 1989, vol. 132, pp. 398–409.
Frankfurter, A., Binder, L.I., and Rebhun, L., Limited tissue distribution of a novel β-tubulin isoform, J. Cell Biol., 1986, vol. 103, p. 273.
Maltman, D.J., Christie, V.B., Collings, J.C., et al., Proteomic profiling of the stem cell response to retinoic acid and synthetic retinoid analogues: identification of major retinoid-inducible proteins, Mol. Biosyst., 2009, vol. 5, no. 5, pp. 458–471.
Nilbratt, M., Friberg, L., Mousavi, M., et al., Retinoic acid and nerve growth factor induce differential regulation of nicotinic acetylcholine receptor subunit expression in SN56 cells, J. Neurosci. Res., 2007, vol. 85, no. 3, pp. 504–514.
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Original Ukrainian Text © V.I. Tsymbalyuk, I.G. Vasil’eva, N.P. Oleksenko, N.G. Chopik, O.I. Tsyubko, O.S. Galanta, 2013, published in Tsitologiya i Genetika, 2013, Vol. 47, No. 3, pp. 54–59.
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Tsymbalyuk, V.I., Vasil’eva, I.G., Oleksenko, N.P. et al. Stimulation of cholinogenesis in human fetal nerve cell cultures. Cytol. Genet. 47, 174–178 (2013). https://doi.org/10.3103/S0095452713030109
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DOI: https://doi.org/10.3103/S0095452713030109