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Neuroscience and Behavioral Physiology

, Volume 50, Issue 1, pp 115–118 | Cite as

The Importance of Extracellular Potassium for Differentiation of Cerebellar Purkinje Cells in Tissue Cultures

  • Yu. D. Stepanenko
  • T. V. Karelina
  • P. A. Abushik
  • D. A. SibarovEmail author
Article
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Despite the fact that cultures of cerebellar granule cells constitute a popular experimental model, the key functional element of the cerebellum, i.e., Purkinje cells (PC), are rarely studied in tissue culture. There are currently serious methodological disagreements relating to the composition of the medium promoting survival of PC and the development of morphology similar to that of in vivo neurons. The development of cerebellar PC in primary tissue cultures at different potassium concentrations was assessed using combinations of vital calcium imaging and immunohistochemical tissue staining for calbindin-28K as a marker for PC. Studies of rat cerebellum were carried out on culture days 7, 14, and 21. Increases in potassium levels in the medium were found to stimulate PC survival, which promoted increases in body size and the development of the dendritic tree of PC. PC demonstrated the intrinsic spontaneous oscillations in free calcium typical for adult PC.

Keywords

Purkinje cells cerebellum primary culture neurons 

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References

  1. 1.
    T. V. Karelina, Yu. D. Stepanenko, P. A. Abushik, et al., “Effects of calcium-activated potassium channel modulators on rat cerebellar Purkinje cell activity,” Acta Naturae, 4, No. 31, 99–107 (2016).Google Scholar
  2. 2.
    J. A. Armengol and C. Sotelo, “Early dendritic development of Purkinje cells in the rat cerebellum. A light and electron microscopic study using axonal tracing in ‘in vitro’ slices,” Dev. Brain Res., 64, No. 1–2, 95–114 (1991).CrossRefGoogle Scholar
  3. 3.
    M. Berry and P. Bradley, “The growth of the dendritic trees of Purkinje cells in the cerebellum of the rat,” Brain Res., 112, No. 1, 1–35 (1976).CrossRefGoogle Scholar
  4. 4.
    F. Boukhtouche, M. Doulazmi, F. Frederic, et al., “RORalpha, a pivotal nuclear receptor for Purkinje neuron survival and differentiation: from development to ageing,” Cerebellum, 5. No. 2, 97–104 (2006).Google Scholar
  5. 5.
    N. Fujikawa, K. Tominaga-Yoshino, M. Okabe, and A., Ogura, “Depolarization dependent survival of cultured mouse cerebellar granule neurons is strain-restrained,” Eur. J. Neurosci., 12, No. 5, 1838–1842 (2000).CrossRefGoogle Scholar
  6. 6.
    J. Hartmann and A. Konnerth, “Determinants of postsynaptic Ca2+ signaling in Purkinje neurons,” Cell Calcium, 37, No. 5 special issue, 459–466 (2005).CrossRefGoogle Scholar
  7. 7.
    P. E. Hockberger, H. Y. Tseng, and J. A. Connor, “Development of rat cerebellar Purkinje cells: electrophysiological properties following acute isolation and in long-term culture,” J. Neurosci., 9, No. 7, 2258–2271 (1989).CrossRefGoogle Scholar
  8. 8.
    B. E. McKay and R. W. Turner, “Physiological and morphological development of the rat cerebellar Purkinje cell,” J. Physiol., 567, No. 3, 829–850 (2005).CrossRefGoogle Scholar
  9. 9.
    M. E. Morrison and C. A. Mason, “Granule neuron regulation of Purkinje cell development: striking a balance between neurotrophin and glutamate signaling,” J. Neurosci., 18, No. 10, 3563–3573 (1998).CrossRefGoogle Scholar
  10. 10.
    J. R. Savidge and D. R. Bristow, “Distribution of Ca2+-permeable AMPA receptors among cultured rat cerebellar granule cells,” Neuroreport, 8, No. 8, 1877–1882 (1997).CrossRefGoogle Scholar
  11. 11.
    D. A. Sibarov, Yu. D. Stepanenko, I. V. Silantiev, et al., “Developmental changes of synaptic and extrasynaptic NMDA receptor expression in rat cerebellar neurons in vitro,” J. Mol. Neurosci., 64, No. 2, 300–311 (2018).CrossRefGoogle Scholar
  12. 12.
    C. Sotelo and I. Dusart, “Intrinsic versus extrinsic determinants during the development of Purkinje cell dendrites,” Neuroscience, 162, No. 3, 589–600 (2009).CrossRefGoogle Scholar
  13. 13.
    M. Yuzaki, D. Forrest, L. M. Verselis, et al., “Functional NMDA receptors are transiently active and support the survival of Purkinje cells in culture,” J. Neurosci., 16, No. 15, 4651–4661 (1996).CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Yu. D. Stepanenko
    • 1
  • T. V. Karelina
    • 1
  • P. A. Abushik
    • 1
  • D. A. Sibarov
    • 1
    Email author
  1. 1.Sechenov Institute of Evolutionary Biochemistry and PhysiologyRussian Academy of SciencesSt. PetersburgRussia

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