Neurochemical Research

, Volume 38, Issue 2, pp 382–388 | Cite as

Rapid Elevation of Calcium Concentration in Cultured Dorsal Spinal Cord Astrocytes by Corticosterone

  • Junwei Zeng
  • Min Li
  • Zhi Xiao
  • Yuanshou Chen
  • Quanzhong Chang
  • Hong Tian
  • Huan Jin
  • Xiaohong Liu
Original Paper


In addition to the classic genomic effects, increasing evidence suggests that GC can generate multiple rapid effects on many tissues and cells through nongenomic pathway. In the present study, the effects of corticosterone (CORT) on the intracellular calcium concentration ([Ca2+]i) in cultured dorsal spinal cord astrocytes were detected with confocal laser scanning microscopy using fluo-4/AM as a calcium fluorescent indicator that could monitor real-time alterations of [Ca2+]i. CORT (0.01–10 μM) caused a rapid increase in [Ca2+]i with a dose-dependent manner in cultured dorsal spinal cord astrocytes. The action of CORT on astrocytic [Ca2+]i was blocked by pertussis toxin (a blocker of G protein activation, 100 ng/ml), but was unaffected by RU38486 (glucocorticoid receptor antagonist, 10 μM). In addition, cycloheximide (protein-synthesis inhibitor, 10 μg/ml) pretreatment could not impair the CORT-evoked [Ca2+]i elevation. Furthermore, Ca2+ mobilization induced by CORT was abolished by chelerythrine chloride (protein kinase C inhibitor, 10 μM), but was not impaired by H89 (protein kinase A inhibitor, 10 μM). These observations suggest that a nongenomic pathways might be involved in the effect of CORT on [Ca2+]i in cultured dorsal spinal cord astrocytes. In addition, our results also raise a possibility that a putative pertussis toxin-sensitive mGCR (G-protein-coupled membrane-bound glucocorticoid receptor) and the downstream activation of protein kinase C may be responsible for CORT-induced Ca2+ mobilization in cultured dorsal spinal cord astrocytes.


Corticosterone Spinal cord Astrocytes Calcium 



This work is supported by the National Natural Science Foundation of China (No.30960126 and 31000497). We are very grateful to the other staff of Department of Physiology. We also thank Guo Luo and Jing-yu Xu (Central Laboratory, Zunyi Medical College) for their technique assistance in laser scanning confocal microscopy.


  1. 1.
    Fuxe K, Härfstrand A, Agnati LF, Yu ZY, Cintra A, Wikström AC, Okret S, Cantoni E, Gustafsson JA (1985) Immunocytochemical studies on the localization of glucocorticoid receptor immunoreactive nerve cells in the lower brain stem and spinal cord of the male rat using a monoclonal antibody against rat liver glucocorticoid receptor. Neurosci Lett 60:1–6PubMedCrossRefGoogle Scholar
  2. 2.
    Ahima RS, Harlan RE (1990) Charting of type II glucocorticoid receptor-like immunoreactivity in the rat central nervous system. Neuroscience 39:579–604PubMedCrossRefGoogle Scholar
  3. 3.
    Yan P, Xu J, Li Q, Chen S, Kim GM, Hsu CY, Xu XM (1999) Glucocorticoid receptor expression in the spinal cord after traumatic injury in adult rats. J Neurosci 19:9355–9363PubMedGoogle Scholar
  4. 4.
    Wang S, Lim G, Zeng Q, Sung B, Ai Y, Guo G, Yang L, Mao J (2004) Expression of central glucocorticoid receptors after peripheral nerve injury contributes to neuropathic pain behaviors in rats. J Neurosci 24:8595–8605PubMedCrossRefGoogle Scholar
  5. 5.
    Wang S, Lim G, Zeng Q, Sung B, Yang L, Mao J (2005) Central glucocorticoid receptors modulate the expression and function of spinal NMDA receptors after peripheral nerve injury. J Neurosci 25:488–495PubMedCrossRefGoogle Scholar
  6. 6.
    Wang S, Lim G, Mao J, Sung B, Yang L, Mao J (2007) Central glucocorticoid receptors regulate the upregulation of spinal cannabinoid-1 receptors after peripheral nerve injury in rats. Pain 131:96–105PubMedCrossRefGoogle Scholar
  7. 7.
    Jackson MJ, Zielke HR, Max SR (1995) Effect of dibutyryl cyclic AMP and dexamethasone on glutamine synthetase gene expression in rat astrocytes in culture. Neurochem Res 20:201–207PubMedCrossRefGoogle Scholar
  8. 8.
    Crossin KL, Tai MH, Krushel LA, Mauro VP, Edelman GM (1997) Glucocorticoid receptor pathways are involved in the inhibition of astrocyte proliferation. Proc Natl Acad Sci USA 94:2687–2692PubMedCrossRefGoogle Scholar
  9. 9.
    Liu Y, Imai H, Sadamatsu M, Tsunashima K, Kato N (2005) Cytokines participate in neuronal death induced by trimethyltin in the rat hippocampus via type II glucocorticoid receptors. Neurosci Res 51:319–327PubMedCrossRefGoogle Scholar
  10. 10.
    Hald A, Nedergaard S, Hansen RR, Ding M, Heegaard AM (2009) Differential activation of spinal cord glial cells in murine models of neuropathic and cancer pain. Eur J Pain 13:138–145PubMedCrossRefGoogle Scholar
  11. 11.
    Wang W, Wang Y, Huang J et al (2008) Temporal changes of astrocyte activation and glutamate transporter-1 expression in the spinal cord after spinal nerve ligation-induced neuropathic pain. Anat Rec (Hoboken) 291:513–518CrossRefGoogle Scholar
  12. 12.
    Lo ¨sel R, Wehling M (2003) Nongenomic actions of steroid hormones. Nat Rev Mol Cell Biol 4:46–56CrossRefGoogle Scholar
  13. 13.
    Liu L, Wang C, Ni X, Sun J (2007) A rapid inhibition of NMDA receptor current by corticosterone in cultured hippocampal neurons. Neurosci Lett 420:245–250PubMedCrossRefGoogle Scholar
  14. 14.
    Yukawa H, Shen J, Harada N, Cho-Tamaoka H, Yamashita T (2005) Acute effects of glucocorticoids on ATP-induced Ca2+ mobilization and nitric oxide production in cochlear spiral ganglion neurons. Neuroscience 130:485–496PubMedCrossRefGoogle Scholar
  15. 15.
    Liu XH, Zeng JW, Zhao YD, Xiao Z, Fang Cq, Ruan HZ (2010) Inhibition of ATP-induced Ca2+ influx by corticosterone in dorsal root ganglion neurons. Neurochem Res 35:804–810PubMedCrossRefGoogle Scholar
  16. 16.
    Simard M, Couldwell WT, Zhang W, Song H, Liu S, Cotrina ML, Goldman S, Nedergaard M (1999) Glucocorticoids-potent modulators of astrocytic calcium signaling. Glia 28:1–12PubMedCrossRefGoogle Scholar
  17. 17.
    Salter MW, Hicks JL (1994) ATP-evoked increases in intracellular calcium in neurons and glia from the dorsal spinal cord. J Neurosci 14:1563–1575PubMedGoogle Scholar
  18. 18.
    Peterson ER, Crain SM (1982) Nerve growth factor attenuates neuro-toxic effects of taxol on spinal cord-ganglion explants from fetal mice. Science 217:377–379PubMedCrossRefGoogle Scholar
  19. 19.
    McDonough SI, Cseresnyés Z, Schneider MF (2000) Origin sites of calcium release and calcium oscillations in frog sympathetic neurons. J Neurosci 20:9059–9070PubMedGoogle Scholar
  20. 20.
    Gee KR, Brown KA, Chen WN, Bishop-Stewart J, Gray D, Johnson I (2000) Chemical and physiological characterization of fluo-4 Ca2+-indicator dyes. Cell Calcium 27:97–106PubMedCrossRefGoogle Scholar
  21. 21.
    Han JZ, Lin W, Chen YZ (2005) Inhibition of ATP-induced calcium influx in HT4 cells by glucocorticoids: involvement of protein kinase A. Acta Pharmacol Sin 26:199–204PubMedCrossRefGoogle Scholar
  22. 22.
    He LM, Zhang CG, Zhou Z, Xu T (2003) Rapid inhibitory effects of corticosterone on calcium influx in rat dorsal root ganglion neurons. Neuroscience 116:325–333PubMedCrossRefGoogle Scholar
  23. 23.
    Qiu J, Wang CG, Huang XY, Chen YZ (2003) Nongenomic mechanism of glucocorticoid inhibition of bradykinin-induced calcium influx in PC12 cells: possible involvement of protein kinase C. Life Sci 72:2533–2542PubMedCrossRefGoogle Scholar
  24. 24.
    Swanson RA, Ying W, Kauppinen TM (2004) Astrocyte influences on ischemic neuronal death. Curr Mol Med 4:193–205PubMedCrossRefGoogle Scholar
  25. 25.
    Carmignoto G, Fellin T (2006) Glutamate release from astrocytes as a non-synaptic mechanism for neuronal synchronization in the hippocampus. J Physiol Paris 99:98–102PubMedCrossRefGoogle Scholar
  26. 26.
    Ni Y, Malarkey EB, Parpura V (2007) Vesicular release of glutamate mediates bidirectional signaling between astrocytes and neurons. J Neurochem 103:1273–1284PubMedCrossRefGoogle Scholar
  27. 27.
    Zeng JW, Liu XH, Zhang JH, Wu XG, Ruan HZ (2008) P2Y1 receptor-mediated glutamate release from cultured dorsal spinal cord astrocytes. J Neurochem 106:2106–2118PubMedCrossRefGoogle Scholar
  28. 28.
    Yang Y, Ge W, Chen Y, Zhang Z, Shen W, Wu C, Poo M, Duan S (2003) Contribution of astrocytes to hippocampal long-term potentiation through release of d-serine. Proc Natl Acad Sci USA 100:15194–15199PubMedCrossRefGoogle Scholar
  29. 29.
    Koizumi S, Fujishita K, Tsuda M, Shigemoto-Mogami Y, Inoue K (2003) Dynamic inhibition of excitatory synaptic transmission by astrocyte-derived ATP in hippocampal cultures. Proc Natl Acad Sci USA 100:11023–11028PubMedCrossRefGoogle Scholar
  30. 30.
    Zeng JW, Liu XH, Zhao YD, Xiao Z, He WJ, Hu ZA, Ruan HZ (2009) Role of P2Y1 receptor in astroglia-to-neuron signaling at dorsal spinal cord. J Neurosci Res 87:2667–2676PubMedCrossRefGoogle Scholar
  31. 31.
    Etgen AM, Martin M, Gilbert R, Lynch G (1980) Characterization of corticosterone-induced protein synthesis in hippocampal slices. J Neurochem 35:598–602PubMedCrossRefGoogle Scholar
  32. 32.
    Morsink MC, Joe ¨ls M, Sarabdjitsingh RA, Meijer OC, De Kloet ER, Datson NA (2006) The dynamic pattern of glucocorticoid receptor-mediated transcriptional responses in neuronal PC12 cells. J Neurochem 99:1282–1298PubMedCrossRefGoogle Scholar
  33. 33.
    Stahn C, Lo ¨wenberg M, Hommes DW, Buttgereit F (2007) Molecular mechanisms of glucocorticoid action and selective glucocorticoid receptor agonists. Mol Cell Endocrinol 275:71–78PubMedCrossRefGoogle Scholar
  34. 34.
    Gagne D, Pons M, Philibert D (1985) RU38486: a potent antiglucocorticoid in vitro and in vivo. J Steroid Biochem 23:247–251PubMedCrossRefGoogle Scholar
  35. 35.
    Vesce S, Rossi D, Brambilla L, Volterra A (2007) Glutamate release from astrocytes in physiological conditions and in neuro-degenerative disorders characterized by neuroinflammation. Int Rev Neurobiol 82:57–71PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Junwei Zeng
    • 1
  • Min Li
    • 1
  • Zhi Xiao
    • 1
  • Yuanshou Chen
    • 1
  • Quanzhong Chang
    • 1
  • Hong Tian
    • 1
  • Huan Jin
    • 1
  • Xiaohong Liu
    • 1
  1. 1.Department of physiologyZunyi Medical CollegeZunyiPeople’s Republic of China

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