Abstract
We have observed the excitability of rat neural progenitor cells (NPCs) and characterized the profile of the voltage-gated ion channels, with the help of the electrophysiological measurements in NPCs in vitro, in order to clarify the electrophysiological property of NPCs. The membrane potential changes of neural progenitors were detected with fluorescent dye bis-(1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC4(3)) by a confocal laser scanning microscope. The changes of fluorescent intensity of DiBAC4(3) stain after KCl stimulation were slight which indicated that the NPCs were inexcitable. Under the current-clamp, spontaneous discharge was not detected and the action potential was failed to be elicited. These findings were consistent with the result from DiBAC4(3) staining. Under the voltage clamp, the NPCs expressed two types of outward K+ currents with no evidence for Na+ currents. An outward delayed rectifier type K+ current and outward transient K+ current were elicited. Our findings demonstrate NPCs’ electrophysiological properties: the electrical inexcitability indicated by the presence of two types of K+ currents and the absence of Na+ current.
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Abrous, D. N., Koehl, M., & Le Moal, M. (2005). Adult neurogenesis: from precursors to network and physiology. Physiological Reviews, 85, 523–569.
Baxter, D. F., Kirk, M., Garcia, A. F., Raimondi, A., Holmqvist, M. H., Flint, K. K., et al. (2002). A novel membrane potential-sensitive fluorescent dye improves cell-based assays for ion channels. Journal of Biomolecular Screening, 7, 79–85.
Berger, T., Schnitzer, J., & Kettenmann, H. (1991). Developmental changes in the membrane current pattern, K+ buffer capacity, and morphology of glial cells in the corpus callosum slice. Journal of Neuroscience, 11, 3008–3024.
Cho, T., Bae, J. H., Choi, H. B., Kim, S. S., McLarnon, J. G., Suh-Kim, H., et al. (2002). Human neural stem cells: electrophysiological properties of voltage-gated ion channels. Neuroreport, 13, 1447–1452.
Encinas, J. M., & Enikolopov, G. (2008). Identifying and quantitating neural stem and progenitor cells in the adult brain. Methods in Cell Biology, 85, 243–272.
Epps, D. E., Wolfe, M. L., & Groppi, V. (1994). Characterization of the steady-state and dynamic fluorescence properties of the potential-sensitive dye bis-(1,3-dibutylbarbituric acid) trimethine oxonol (Dibac4(3)) in model systems and cells. Chemistry and Physics of Lipids, 69, 137–150.
Feldman, D. H., Thinschmidt, J. S., Peel, A. L., Papke, R. L., & Reier, P. J. (1996). Differentiation of ionic currents in CNS progenitor cells: dependence upon substrate attachment and epidermal growth factor. Experimental Neurology, 140, 206–217.
Giles, W., Nakajima, T., Ono, K., & Shibata, E. F. (1989). Modulation of the delayed rectifier K+ current by isoprenaline in bull-frog atrial myocytes. Journal of Physiology, 415, 233–249.
Hamill, O. P., Marty, A., Neher, E., Sakmann, B., & Sigworth, F. J. (1981). Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Archiv, 391, 85–100.
Hausser, M., Raman, I. M., Otis, T., Smith, S. L., Nelson, A., du Lac, S., et al. (2004). The beat goes on: spontaneous firing in mammalian neuronal microcircuits. Journal of Neuroscience, 24, 9215–9219.
Herberth, B., Pataki, A., Jelitai, M., Schlett, K., Deak, F., Spat, A., et al. (2002). Changes of KCl sensitivity of proliferating neural progenitors during in vitro neurogenesis. Journal of Neuroscience Research, 67, 574–582.
Hogg, R. C., Chipperfield, H., Whyte, K. A., Stafford, M. R., Hansen, M. A., Cool, S. M., et al. (2004). Functional maturation of isolated neural progenitor cells from the adult rat hippocampus. European Journal of Neuroscience, 19, 2410–2420.
Klee, R., Ficker, E., & Heinemann, U. (1995). Comparison of voltage-dependent potassium currents in rat pyramidal neurons acutely isolated from hippocampal regions CA1 and CA3. Journal of Neurophysiology, 74, 1982–1995.
Korotzer, A. R., & Cotman, C. W. (1992). Voltage-gated currents expressed by rat microglia in culture. Glia, 6, 81–88.
Liebau, S., Propper, C., Bockers, T., Lehmann-Horn, F., Storch, A., Grissmer, S., et al. (2006). Selective blockage of Kv1.3 and Kv3.1 channels increases neural progenitor cell proliferation. Journal of Neurochemistry, 99, 426–437.
Neusch, C., Weishaupt, J. H., & Bahr, M. (2003). Kir channels in the CNS: emerging new roles and implications for neurological diseases. Cell & Tissue Research, 311, 131–138.
Nichols, J., Zevnik, B., Anastassiadis, K., Niwa, H., Klewe-Nebenius, D., Chambers, I., et al. (1998). Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell, 95, 379–391.
Park, K. S., Jung, K. H., Kim, S., Kim, K. S., Choi, M. R., Kim, Y., et al. (2007). Functional expression of ion channels in mesenchymal stem cells derived from umbilical cord vein. Stem Cells, 25, 2044–2052.
Patil, N., Cox, D. R., Bhat, D., Faham, M., Myers, R. M., & Peterson, A. S. (1995). A potassium channel mutation in weaver mice implicates membrane excitability in granule cell differentiation. Nature Genetics, 11, 126–129.
Piper, D. R., Mujtaba, T., Rao, M. S., & Lucero, M. T. (2000). Immunocytochemical and physiological characterization of a population of cultured human neural precursors. Journal of Neurophysiology, 84, 534–548.
Reyes-Haro, D., Miledi, R., & Garcia-Colunga, J. (2005). Potassium currents in primary cultured astrocytes from the rat corpus callosum. Journal of Neurocytology, 34, 411–420.
Stewart, R. R., Zigova, T., & Luskin, M. B. (1999). Potassium currents in precursor cells isolated from the anterior subventricular zone of the neonatal rat forebrain. Journal of Neurophysiology, 81, 95–102.
Tse, F. W., Fraser, D. D., Duffy, S., & MacVicar, B. A. (1992). Voltage-activated K+ currents in acutely isolated hippocampal astrocytes. Journal of Neuroscience, 12, 1781–1788.
Wonderlin, W. F., & Strobl, J. S. (1996). Potassium channels, proliferation and G1 progression. Journal of Membrane Biology, 154, 91–107.
Yamada, A., Gaja, N., Ohya, S., Muraki, K., Narita, H., Ohwada, T., et al. (2001). Usefulness and limitation of DiBAC4(3), a voltage-sensitive fluorescent dye, for the measurement of membrane potentials regulated by recombinant large conductance Ca2+-activated K+ channels in HEK293 cells. Japanese Journal of Pharmacology, 86, 342–350.
Yuan, X., Eisen, A. M., McBain, C. J., & Gallo, V. (1998). A role for glutamate and its receptors in the regulation of oligodendrocyte development in cerebellar tissue slices. Development, 125, 2901–2914.
Zhang, M., Guo, Y., Jiang, L., & Liu, G. X. (2007). The morphology and membrane potential of primary cultured rat hippocampal neurons with serum-free media. Chinese Journal of Cell Biology, 29, 131–134 (in Chinese).
Acknowledgements
We thank Dr. Congjian Zhao (Max-Delbrück-Center for Molecular Medicine, Berlin-Buch) for correcting the English writing of the manuscript and the editor and reviewers for their valuable comments and suggestions that greatly improved the presentation of this paper. This work was supported by grants from the National Natural Science Foundation of China (No. 30672217) and the Science Foundation of Chongqing Health Bureau (No. 06-2-158).
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Li, T., Jiang, L., Chen, H. et al. Characterization of Excitability and Voltage-gated Ion Channels of Neural Progenitor Cells in Rat Hippocampus. J Mol Neurosci 35, 289–295 (2008). https://doi.org/10.1007/s12031-008-9065-7
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DOI: https://doi.org/10.1007/s12031-008-9065-7