Abstract
Retinoic acid differentiated SH-SY5Y cells exhibit only a high-threshold-activated (−30 to −20 mV) whole cell calcium channel current. When barium was used as the charge carrier, the high-threshold-activated current showed bi-exponential inactivation kinetics during a 500 ms voltage step from −90 to +10mV. The time constants of inactivation were approximately 75 and 750 ms. The fast inactivating component was more sensitive than the slow inactivating component to steady-state inactivation at depolarized holding potentials. The calcium channel current was inhibited by externally applied cadmium (10–300 μM) and gadolinium (10–30 μM) as well as by high concentrations of nickel and cobalt, ω Conus toxin (1 μM) irreversibly blocked the calcium channel current. However, the dihydropyridine agonist, BAY K 8644 (3–10 μM) and antagonists, nifedipine (3–10 μM) and nimodipine (10 μM) did not affect either component of the calcium channel current. Agents which blocked the calcium channel current did not exhibit any selectivity for the fast inactivating over the slow inactivating component of the current. These results indicate that whilst the calcium channel current recorded in differentiated SH-SY5Y cells can be classified on the basis of the blocking agents as being of the N type, the current shows more than one form of inactivation.
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References
Åkerman KEO, Scott IG, Andersson LC (1984) Functional differentiation of a human ganglion cell derived neuroblastoma cell line SH-SY5Y induced by a phorbol ester (TPA). Neurochem Int 6:77–80
Aosaki T, Kasai H (1989) Characterization of two kinds of high voltage-activated Ca-channel currents in chick sensory neurons: differential sensitivity to dihydropyridines and ω Conotoxin CVIA. Pflügers Arch 414:150–156
Armstrong D, Eckert R (1987) Voltage-activated calcium channels that must be phosphorylated to respond to membrane depolarization. Proc Natl Acad Sci USA 84:2518–1522
Biedler JL, Roffler-Tarlov S, Schachner M, Freedman LS (1978) Multiple neurotransmitter synthesis by human neuroblastoma cell lines and clones. Cancer Res 38:3751–3757
Bosma M, Sidell N (1988) Retinoic acid inhibits Ca++ currents and cell proliferation in a B-lymphocyte cell line. J Cell Physiol 135:317–323
Docherty RJ (1988) Gadolinium selectively blocks a component of calcium current in rodent neuroblastoma x glioma hybrid (NG108-15) cells. J Physiol (Lond) 398:33–48
Doerner D, Pither TA, Alger BE (1988) Protein kinase C activators block specific calcium and potassium current components in isolated hippocampal neurons. J Neurosci 8:4069–4078
Fox AP, Nowycky MC, Tsien RW (1987) Kinetic and pharmacological properties distinguishing three types of calcium currents in chick sensory neurones. J Physiol (Lond) 394:149–172
Gross RA, MacDonald RL (1989a) Cyclic AMP selectively reduces the N-type calcium current component of mouse sensory neurones in culture by enhancing inactivation. J Neurophysiol 61:97–105
Gross RA, MacDonald RL (1989b) Activators of protein kinase C selectively enhance inactivation of a calcium current component of cultured sensory neurons in a pertussis toxin-sensitive manner. J Neurophysiol 61:1259–1269
Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100
Henderson G, Seward EP (1990) Inhibition of an N-like calcium channel current by μ opioid receptor activation in the human neuroblastoma cell line SH-SY5Y. J Physiol (Lond) 422:19P
Hirning LD, Fox AP, McClesskey EW, Miller RJ, Olivera BM, Thayer SA, Miller RJ, Tsien RW (1988) Dominant role of N-type calcium channels in evoked release of norepinephrine from sympathetic neurons. Science 239:57–61
Ino M, Cole GM, Timaras PS (1986) Tyrosine hydroxylase and monoamine oxidase-A activity increases in differentiating human neuroblastoma after elimination of dividing cells. Dev Brain Res 30:120–123
Kasai H, Aosaki T (1988) Divalent cation dependent inactivation of the high-voltage-activated Ca-channel current in chick sensory neurons. Pflügers Arch 411:695–697
Klinz FJ, Yu VC, Sadée W, Costa T (1987) Differential expression of α subunits of G-proteins in human neuroblastoma-derived cell clones. FEBS Lett 224:43–48
Kongsamut S, Lipscombe D, Tsien RW (1989) The N-type Ca channel in frog sympathetic neurons and its role in α adrenergic modulation of transmitter release. Ann NY Acad Sci 560:312–333
Kuramoto T, Werrbach-Perez K, Perez-Polo JR, Haber B (1981) Membrane properties of a human neuroblastoma II. Effects of differentiation. J Neurosci Res 6:441–449
Lambert DG, Whitman EM, Baird JG, Nahorski SR (1990) Different mechanism of Ca2+ entry induced by depolarization and muscarinic receptor stimulation in SH-SY5Y human neuroblastoma cells. Mol Brain Res 8:263–266
Lipscombe D, Kongsamut S, Tsien RW (1989) α-Adrenergic inhibiton of sympathetic neurotransmitter release mediated by modulation of N-type calcium channel gating. Nature 340:639–642
Llinas RR, Sugimori M, Cherksey B (1989) Voltage-dependent calcium conductances in mammalian neurons. Ann NY Acad Sci 560:103–111
Narahashi T, Tsunoo A, Yoshii M (1987) Characterization of two types of calcium channels in mouse neuroblastoma cells. J Physiol (Lond) 383:231–249
Nowycky MC, Fox AP, Tsien RW (1985) Three types of neuronal calcium channels with different calcium agonist sensitivity. Nature 316:440–443
Pålman S, Odelsted L, Larsson E, Grotte G, Nilsson K (1981) Phenotypic changes of human neuroblastoma cells in culture induced by 12-0-tetradeconyl-phorbol-13-acetate. Int J Cancer 28:583–589
Perez-Polo JR, Werbach-Perez K, Tiffany-Castiglioni E (1979) A humn clonal cell line model of differentiating neurons. Dev Biol 71:341–355
Plummer MR, Logothetis DE, Hess P (1989) Elementary properties and pharmacological sensitivities of calcium channels in mammalian peripheral neurons. Neuron 2:1453–1463
Rane SG, Holz GG, Dunlap K (1987) Dihydropyridine inhibition of neuronal calcium current and substance P release. Pflügers Arch 409:361–366
Scott IG, Åkerman KEO, Heikkilå JE, Kaila K, Andersson LC (1986) Development of a neuronal phenotype in differentiating ganglion cell-derived human neuroblastoma cells. J Cell Physiol 128:285–292
Seward EP, Henderson G, Sadée W (1989) Electrophysiological properties of retinoic acid differentiated human neuroblastoma (SH-SY5Y) cells. Eur J Neurosci Suppl 1:77.14
Spitzer NC (1979) Ion channels in development. Annu Rev Neurosci 2:363–397
Streit J, Lux HD (1988) Effects of nerve growth factor on calcium currents in PC12 cells. Soc Neurosci Abstr 14:55.14
Swandulla D, Armstrong CM (1988) Fast-deactivating calcium channels in chick sensory neurons. J Gen Physiol 92:197–218
Wanke E, Ferroni A, Malgaroli A, Pozzan T, Meldosi J (1987) Activation of a muscarinic receptor selectively inhibits a rapidly inactivated Ca2+ current in rat sympathetic neurons. Proc Natl Acad Sci USA 84:4313–4317
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Seward, E.P., Henderson, G. Characterization of two components of the N-like, high-threshold-activated calcium channel current in differentiated SH-SY5Y cells. Pflugers Arch. 417, 223–230 (1990). https://doi.org/10.1007/BF00370703
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DOI: https://doi.org/10.1007/BF00370703