SpringerLink
Log in

CNTF-Treated Astrocyte Conditioned Medium Enhances Large-Conductance Calcium-Activated Potassium Channel Activity in Rat Cortical Neurons

  • Original Paper
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

Seizure activity is linked to astrocyte activation as well as dysfunctional cortical neuron excitability produced from changes in calcium-activated potassium (KCa) channel function. Ciliary neurotrophic factor-treated astrocyte conditioned medium (CNTF-ACM) can be used to investigate the peripheral effects of activated astrocytes upon cortical neurons. However, CNTF-ACM’s effect upon KCa channel activity in cultured cortical neurons has not yet been investigated. Whole-cell patch clamp recordings were performed in rat cortical neurons to evaluate CNTF-ACM’s effects upon charybdotoxin-sensitive large-conductance KCa (BK) channel currents and apamin-sensitive small-conductance KCa (SK) channel current. Biotinylation and RT-PCR were applied to assess CNTF-ACM’s effects upon the protein and mRNA expression, respectively, of the SK channel subunits SK2 and SK3 and the BK channel subunits BKα1 and BKβ3. An anti-fibroblast growth factor-2 (FGF-2) monoclonal neutralizing antibody was used to assess the effects of the FGF-2 component of CNTF-ACM. CNTF-ACM significantly increased KCa channel current density, which was predominantly attributable to gains in BK channel activity (p < 0.05). CNTF-ACM produced a significant increase in BKα1 and BKβ3 expression (p < 0.05) but had no significant effect upon SK2 or SK3 expression (p > 0.05). Blocking FGF-2 produced significant reductions in KCa channel current density (p > 0.05) as well as BKα1 and BKβ3 expression in CNTF-ACM-treated neurons (p > 0.05). CNTF-ACM significantly enhances BK channel activity in rat cortical neurons and that FGF-2 is partially responsible for these effects. CNTF-induced astrocyte activation results in secretion of neuroactive factors which may affect neuronal excitability and resultant seizure activity in mammalian cortical neurons.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Fisher RS, Acevedo C, Arzimanoglou A, Bogacz A, Cross JH, Elger CE, Engel J, Forsgren L, French JA, Glynn M (2014) ILAE official report: a practical clinical definition of epilepsy. Epilepsia 55:475–482

    Article  PubMed  Google Scholar 

  2. Nevalainen O, Simola M, Ansakorpi H, Raitanen J, Artama M, Isojärvi J, Auvinen A (2015) Epilepsy, excess deaths and years of life lost from external causes. Eur J Epidemiol 30:1–9

  3. N’Gouemo P (2014) BKCa channel dysfunction in neurological diseases. Front Physiol 5:18–19

  4. Sarafian TA, Montes C, Imura T, Qi J, Coppola G, Geschwind DH, Sofroniew MV (2010) Disruption of astrocyte STAT3 signaling decreases mitochondrial function and increases oxidative stress in vitro. PLoS One 5:e9532

    Article  PubMed  PubMed Central  Google Scholar 

  5. Pekny M, Wilhelmsson U, Pekna M (2014) The dual role of astrocyte activation and reactive gliosis. Neurosci Lett 565:30–38

    Article  CAS  PubMed  Google Scholar 

  6. Pasquin S, Sharma M, Gauchat J-F (2015) Ciliary neurotrophic factor (CNTF): new facets of an old molecule for treating neurodegenerative and metabolic syndrome pathologies. Cytokine Growth Factor Rev 30:18–19

  7. Albrecht PJ, Enterline JC, Cromer J, Levison SW (2007) CNTF-activated astrocytes release a soluble trophic activity for oligodendrocyte progenitors. Neurochem Res 32:263–271

    Article  CAS  PubMed  Google Scholar 

  8. Wang X, Zheng H, Liu C, Zhu C, Wang W, Li Z (2008) Ciliary neurotrophic factor-treated astrocyte conditioned medium regulates the L-type calcium channel activity in rat cortical neurons. Neurochem Res 33:826–832

    Article  CAS  PubMed  Google Scholar 

  9. Holm NR, Christophersen P, Olesen SP, Gammeltoft S (1997) Activation of calcium-dependent potassium channels in rat brain neurons by neurotrophin-3 and nerve growth factor. Proc Natl Acad Sci 94:1002–1006

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Baldelli P, Forni PE, Carbone E (2000) BDNF, NT-3 and NGF induce distinct new Ca2+ channel synthesis in develo** hippocampal neurons. Eur J Neurosci 12:4017–4032

    Article  CAS  PubMed  Google Scholar 

  11. Stocker M, Krause M, Pedarzani P (1999) An apamin-sensitive Ca2+-activated K+ current in hippocampal pyramidal neurons. Proc Natl Acad Sci 96:4662–4667

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Li W, Gao SB, Lv CX, Wu Y, Guo ZH, Ding JP, Xu T (2007) Characterization of voltage-and Ca2+-activated K+ channels in rat dorsal root ganglion neurons. J Cell Physiol 212:348–357

    Article  CAS  PubMed  Google Scholar 

  13. Sun X, Gu XQ, Haddad GG (2003) Calcium influx via L-and N-type calcium channels activates a transient large-conductance Ca2+-activated K+ current in mouse neocortical pyramidal neurons. J Neurosci 23:3639–3648

    CAS  PubMed  Google Scholar 

  14. Chen SR, Cai YQ, Pan HL (2009) Plasticity and emerging role of BKCa channels in nociceptive control in neuropathic pain. J Neurochem 110:352–362

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Joiner WJ, Khanna R, Schlichter LC, Kaczmarek LK (2001) Calmodulin regulates assembly and trafficking of SK4/IK1 Ca2+-activated K+ channels. J Biol Chem 276:37980–37985

    CAS  PubMed  Google Scholar 

  16. Dominguez R, Micevych P (2010) Estradiol rapidly regulates membrane estrogen receptor α levels in hypothalamic neurons. J Neurosci 30:12589–12596

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Yang J, Krishnamoorthy G, Saxena A, Zhang G, Shi J, Yang H, Delaloye K, Sept D, Cui J (2010) An epilepsy/dyskinesia-associated mutation enhances BK channel activation by potentiating Ca2+ sensing. Neuron 66:871–883

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Wang B, Rothberg BS, Brenner R (2009) Mechanism of increased BK channel activation from a channel mutation that causes epilepsy. J Gen Physiol 133:283–294

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Du W, Bautista JF, Yang H, Diez-Sampedro A, You S-A, Wang L, Kotagal P, Lüders HO, Shi J, Cui J (2005) Calcium-sensitive potassium channelopathy in human epilepsy and paroxysmal movement disorder. Nat Genet 37:733–738

    Article  CAS  PubMed  Google Scholar 

  20. Shruti S, Urban-Ciecko J, Fitzpatrick JA, Brenner R, Bruchez MP, Barth AL (2012) The brain-specific β4 subunit downregulates BK channel cell surface expression. PLoS One 7:e33429

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Brenner R, Chen QH, Vilaythong A, Toney GM, Noebels JL, Aldrich RW (2005) BK channel β4 subunit reduces dentate gyrus excitability and protects against temporal lobe seizures. Nat Neurosci 8:1752–1759

    Article  CAS  PubMed  Google Scholar 

  22. Lorenz S, Heils A, Kasper JM, Sander T (2007) Allelic association of a truncation mutation of the KCNMB3 gene with idiopathic generalized epilepsy. Am J Med Genet Part B Neuropsychiatr Genet 144:10–13

    Article  Google Scholar 

  23. Hu S, Labuda MZ, Pandolfo M, Goss GG, McDermid HE, Ali DW (2003) Variants of the KCNMB3 regulatory subunit of maxi BK channels affect channel inactivation. Physiol Genomics 15:191–198

    Article  CAS  PubMed  Google Scholar 

  24. Young KL, Villar D, Carson TL, Imerman PM, Moore RA, Bottoff MR (2003) Tremorgenic mycotoxin intoxication with penitrem A and roquefortine in two dogs. J Am Vet Med Assoc 222:52–53

    Article  PubMed  Google Scholar 

  25. Szebenyi G, Dent EW, Callaway JL, Seys C, Lueth H, Kalil K (2001) Fibroblast growth factor-2 promotes axon branching of cortical neurons by influencing morphology and behavior of the primary growth cone. J Neurosci 21:3932–3941

    CAS  PubMed  Google Scholar 

  26. Liu Z, Holmes GL (1997) Basic fibroblast growth factor-induced seizures in rats. Neurosci Lett 233:85–88

    Article  CAS  PubMed  Google Scholar 

  27. Zucchini S, Buzzi A, Barbieri M, Rodi D, Paradiso B, Binaschi A, Coffin JD, Marzola A, Cifelli P, Belluzzi O (2008) Fgf-2 overexpression increases excitability and seizure susceptibility but decreases seizure-induced cell loss. J Neurosci 28:13112–13124

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Davies PJ, Ireland DR, McLachlan EM (1996) Sources of Ca2+ for different Ca(2+)-activated K+ conductances in neurones of the rat superior cervical ganglion. J Physiol 495:353

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Sah P (1995) Different calcium channels are coupled to potassium channels with distinct physiological roles in vagal neurons. Proc R Soc Lond B Biol Sci 260:105–111

    Article  CAS  Google Scholar 

  30. Alvina K, Khodakhah K (2008) Selective regulation of spontaneous activity of neurons of the deep cerebellar nuclei by N-type calcium channels in juvenile rats. J Physiol 586:2523–2538

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant No. 81171117), the Scientific Research Starting Foundation for the Returned Overseas Chinese Scholars (Chinese Ministry of Education, Grant No. 44), and the Science and Technology Program of Anhui Province (a Key Laboratory Project, Grant No. 1506c085014).

Author Contributions

X. J. W.: Conceived and designed the study. M. Q. S., H. L. L., H. B. X., and H. T. W: Performed the experiments. X. J. W.: Analyzed the data and drafted the manuscript

Author information

Authors and Affiliations

  1. Department of Respiration, Anhui Clinical and Preclinical Key Laboratory of Respiratory Disease, The First Affiliated Hospital of Bengbu Medical College, No. 287 Changhuai Road, Bengbu, Anhui, 233000, China

    **ao**g Wang

  2. Department of Histology and Embryology, Bengbu Medical College, Bengbu, Anhui, China

    Meiqun Sun

  3. Department of Gynecological Oncology, The First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui, China

    Hongli Liu

  4. Department of Endocrinology and Metabolism, Shanghai Jiaotong University Affiliated First People’s Hospital, Shanghai, China

    Huanbai Xu

  5. Department of Immunology, Bengbu Medical College, Bengbu, Anhui, China

    Hongtao Wang

Authors
  1. Meiqun Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hongli Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Huanbai Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hongtao Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. **ao**g Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to **ao**g Wang.

Ethics declarations

Conflict of interest

None.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sun, M., Liu, H., Xu, H. et al. CNTF-Treated Astrocyte Conditioned Medium Enhances Large-Conductance Calcium-Activated Potassium Channel Activity in Rat Cortical Neurons. Neurochem Res 41, 1982–1992 (2016). https://doi.org/10.1007/s11064-016-1910-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11064-016-1910-4

Keywords

Search

Navigation