The Korean Journal of Physiology and Pharmacology

The Korean Society of Pharmacology (대한약리학회)
권호 표지

Calcium Ion Dynamics after Dexamethasone Treatment in Organotypic Cultured Hippocampal Slice

  • Chae, Hee-Jung (Department of Medical Science, Graduate School of East-West Medicine Science, Kyunghee University) ;
  • Kang, Tong-Ho (Department of Medical Science, Graduate School of East-West Medicine Science, Kyunghee University) ;
  • Park, Ji-Ho (Department of Medical Science, Graduate School of East-West Medicine Science, Kyunghee University)
  • Published : 2005.12.21
Download PDF
⟨ Previous Next ⟩

Abstract

It is imperative to analyse brain injuries directly in real time, so as to find effective therapeutic compounds to protect brain injuries by stress. We established a system which could elucidate the real time $Ca^{2+}$ dynamics in an organotypic cultured hippocampal slice by the insults of artificial stress hormone, dexamethasone. The real time $Ca^{2+}$ dynamics could continuously be detected in cornus ammonis 3 (CA3) of the organotypic hippocampus for 8 hours under confocal microscopy. When dexamethasone concentration was increased, the $Ca^{2+}$ was also increased in a dose dependent manner at $1{\sim}100{\mu}M$ concentrations. Moreover, when the organotypic cultured hippocampal slice was treated with a glutamate receptor antagonist together with dexamethasone, the real time $Ca^{2+}$ dynamics were decreased. Furthermore, we confirmed by PI uptake study that glutamate receptor antagonist reduced the hippocampal tissue damage caused by dexamethasone treatment. Therefore, our new calcium ion dynamics system in organotypic cultured hippocampal slice after dexamethasone treatment could provide real time analysis method for investigation of brain injuries by stress.

Keywords

References

  1. Andreas B, Robin ZN, Hans-Peter S, Joachim WD. Calcium release from intracellular stores in rodent astrocytes and neurons in situ. Cell Calcium 35: 47-58, 2004 https://doi.org/10.1016/S0143-4160(03)00171-4
  2. Berridge MJ, Bootman MD, Lipp P. Calcium - a life and death signal. Nature 395: 645-648, 1998 https://doi.org/10.1038/27094
  3. Berridge MJ, Lipp P, Bootman MD. The versatility and universality of calcium signaling. Nat Rev Mol Cell Biol 1: 11-21, 2000
  4. Christian L. Monitoring neuronal calcium signaling using a new method for ratiometric confocal calcium imaging. Cell Calcium 34: 295-303, 2003 https://doi.org/10.1016/S0143-4160(03)00105-2
  5. Chrles LR, Andrew HM. When not enough is too much: the role of insufficient glucocorticoid signaling in the pathophysiology of stress-related disorders. Am J Psychiatry 160: 1554-1565, 2003 https://doi.org/10.1176/appi.ajp.160.9.1554
  6. Ciriza I, Azcoitia I, Garcia-Segura LM. Reduced progesterone metabolites protect rat hippocampal neurones from kainic acid excitotoxicity in vivo. J Neuroendocrinol 16: 58-63, 2004 https://doi.org/10.1111/j.1365-2826.2004.01121.x
  7. Colin B. Cellular calcium imaging: so, what's new? Trend In Cell Biology 10: 451-457, 2000 https://doi.org/10.1016/S0962-8924(00)01799-2
  8. David GL. Calcium signaling protocols. 1st ed. Humana Press, Leicester UK, 1999
  9. Elicia ME, Robert MS. Corticosterone impairs hippocampal neuronal calcium regulationpossible mediating mechanisms. Brain Res 602: 84-90, 1993 https://doi.org/10.1016/0006-8993(93)90245-I
  10. Fabienne R, Anne-Marie B, Pierre B. Dynamic study of cell mechanical and structural responses to rapid changes of calcium level. Cell Motility and Cytoskeleton 45: 93-105, 2000 https://doi.org/10.1002/(SICI)1097-0169(200002)45:2<93::AID-CM2>3.0.CO;2-Z
  11. Gee KR, Brown KA, Chen W-NU, Bishop-Stewart J, Gray D, Johnson I. Chemical and physiological characterization of fluo-4 $Ca^{2+}$-indicator dyes. Cell Calcium 27: 97-106, 2000 https://doi.org/10.1054/ceca.1999.0095
  12. Haugland RP. Handbook of fluorescent probes and research products. 9th ed. Molecular Probes, USA, 2002
  13. Hua W, Erik BM, Sunjara V, Steven ER, Wen-hong L. $Ca^{2+}$ dependent glutamate release involves two classes of endoplasmic reticulum $Ca^{2+}$ stores in astrocytes. J Neurosci Res 76: 86-97, 2004 https://doi.org/10.1002/jnr.20061
  14. Joels M. Corticosteroid actions in the hippocampus. J Neuroendo 13: 657-669, 2001 https://doi.org/10.1046/j.1365-2826.2001.00688.x
  15. Kloet ER. Hormones, brain and stress. Endocrine Regulations 37: 51-68, 2003
  16. Kobayashi T, Yamada T, Okada Y. The levels of adenosine and its metabolites in the guinea pig and rat brain during complete ischemia-in vivo study. Brain Res 787: 211-219, 1998 https://doi.org/10.1016/S0006-8993(97)01481-9
  17. Laake JH, Haug FM, Wieloch T, Ottersen OP. A simple in vitro model of ischemia based on hippocampal slice cultures and propidium iodide fluorescence. Brain Res Protoc 4: 173-184, 1999 https://doi.org/10.1016/S1385-299X(99)00021-5
  18. Lee AL, Ogle WO, Sapolsky RM. Stress and depression: possible links to neuron death in the hippocampus. Bipolar Disord 4: 117-128, 2002 https://doi.org/10.1034/j.1399-5618.2002.01144.x
  19. Macklis JD, Madison RD. Progressive incorporation of propidium iodide in cultured mouse neurons correlates with declining electrophysiological status: a fluorescence scale of membrane integrity. J Neurosci Methods 31: 43-46, 1990 https://doi.org/10.1016/0165-0270(90)90007-3
  20. McEwen BS, Sapolsky RM. Stress and cognitive function. Curr Opin Neurobiol 5: 205-216, 1995 https://doi.org/10.1016/0959-4388(95)80028-X
  21. Meijer OC, Kalkhoven E, Laan S, Steenbergen PJ, Houtman SH, Dijkmans TF, Pearce D, de Kloet ER. Steroid receptor coactivator- 1 splice variants differentially affect corticosteroid receptor signaling. Endocrinology 24: 1-48, 2004
  22. Saino T, Satoh Y. Application of real-time confocal laser scanning microscopy to observe living cells in tissue specimens. J Electron Microsc 53: 49-56, 2004 https://doi.org/10.1093/jmicro/53.1.49
  23. Schuster S, Marhl M, Hofer T. Modeling of simple and complex calcium oscillations. Eur J Biochem 269: 1333-1355, 2002 https://doi.org/10.1046/j.0014-2956.2001.02720.x
  24. Stephen AS, Whitaker M. Confocal laser scanning microscopy of calcium dynamics in living cells. Microscopy Reseach and Technique 46: 356-369, 1999 https://doi.org/10.1002/(SICI)1097-0029(19990915)46:6<356::AID-JEMT4>3.0.CO;2-6
  25. Stoppini LP, Buchs A, Muller D. A simple method for organotypic cultures of nervous tissue. J Neurosci Methods 37: 173-193, 1991 https://doi.org/10.1016/0165-0270(91)90128-M
  26. Supko DE, Johnston MV. Dexamethasone potentiates NMDA receptormediated neuronal injury in the postnatal rat. Eur J Pharmacol 270: 105-113, 1994
  27. Takahashi A, Camacho P, Lechleiter JD, Herman B. Measurement of intracellular calcium. Physiol Rev 79: 1089-1125, 1999
  28. Thomas D, Tovey SC, Collins TJ, Bootman MD, Berridge MJ, Lipp P. A comparison of fluorescent $Ca^{2+}$ indicator properties and their use in measuring elementary and global $Ca^{2+}$ signals. Cell Calcium 28: 213-223, 2000 https://doi.org/10.1054/ceca.2000.0152